UBC Theses and Dissertations

UBC Theses Logo

UBC Theses and Dissertations

Preputial glands of the albino mouse. Still, Susan 1971

Your browser doesn't seem to have a PDF viewer, please download the PDF to view this item.

Item Metadata

Download

Media
831-UBC_1971_A6_7 S85.pdf [ 15.36MB ]
Metadata
JSON: 831-1.0101849.json
JSON-LD: 831-1.0101849-ld.json
RDF/XML (Pretty): 831-1.0101849-rdf.xml
RDF/JSON: 831-1.0101849-rdf.json
Turtle: 831-1.0101849-turtle.txt
N-Triples: 831-1.0101849-rdf-ntriples.txt
Original Record: 831-1.0101849-source.json
Full Text
831-1.0101849-fulltext.txt
Citation
831-1.0101849.ris

Full Text

THE PREPUTIAL  GLANDS  OF THE ALBINO MOUSE  b  SUSAN  y  STILL  B. S c . , D a l h o u s i e  University,  1967  A THESIS SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE i n the Department of Zoology  We a c c e p t t h i s t h e s i s required standard  as c o n f o r m i n g  to the  THE UNIVERSITY OF BRITISH COLUMBIA May, 1971  In  presenting  this  an a d v a n c e d  degree  the  shall  I  Library  f u r t h e r agree  for  scholarly  by  his  of  this  written  thesis at  it  purposes  for  freely  permission may  representatives. thesis  partial  the U n i v e r s i t y  make  that  in  financial  is  Columbia,  British  by  for  gain  Columbia  shall  the  that  not  requirements I  agree  r e f e r e n c e and copying  t h e Head o f  understood  of  The U n i v e r s i t y o f B r i t i s h V a n c o u v e r 8, Canada  of  for extensive  permission.  Department  of  available  be g r a n t e d  It  fulfilment  of  be a l l o w e d  or  that  study.  this  thesis  my D e p a r t m e n t  copying  for  or  publication  without  my  ABSTRACT  The Mus  p a i r e d p r e p u t i a l g l a n d s o f t h e a l b i n o mouse,  musculus,  ( S w i s s s t r a i n ) were s t u d i e d w i t h r e g a r d t o  h i s t o l o g y , p h y s i o l o g y and b a s i c h i s t o c h e m i s t r y . F o r t h e tological  s t u d y o f t h e d e v e l o p m e n t and  subsequent  a g i n g of  t h e p r e p u t i a l g l a n d , t h e g l a n d s of male and f e m a l e d i f f e r e n t ages, were u s e d .  mice o f  r a n g i n g f r o m newborn t o 12 months o l d ,  I n o r d e r to study the e f f e c t s of androgens  e s t r o g e n s upon the m a c r o s c o p i c  and  and m i c r o s c o p i c s t r u c t u r e  t h e p r e p u t i a l g l a n d , w h i t e S w i s s m i c e were d i v i d e d f o u r m a j o r g r o u p s ( a c c o r d i n g t o age which were, i n t u r n , d i v i d e d i n t o for  his-  and  of  into  s t a t u s ) each  of  s u b g r o u p s a, b , c_, and  d  t r e a t m e n t ; mice i n s u b g r o u p s a and b r e c e i v e d d a i l y  subcutaneous  i n j e c t i o n s of t e s t o s t e r o n e p r o p i o n a t e and  of  e s t r a d i o l benzoate  r e s p e c t i v e l y ; mice i n subgroup £ r e c e i v e d  daily  i n j e c t i o n s of t h e v e h i c l e , sesame o i l ;  subcutaneous  mice i n s u b g r o u p d s e r v e d as c o n t r o l s .  There w e r e , o f  male and f e m a l e  Routine h i s t o c h e m i c a l  tests for lipids  mice i n e a c h g r o u p i n g .  and p r o t e i n s were c a r r i e d o u t on  e x c i s e d p r e p u t i a l g l a n d s of w e a n l i n g and of  both sexes, i n order to l o c a l i z e  course,  the  of young a d u l t mice,  and t o i d e n t i f y  the  s e c r e t o r y p r o d u c t s of the a c i n a r c e l l s . H i s t o l o g i c a l l y and p h y s i o l o g i c a l l y response of  t o a n d r o g e n s and  t h e mouse a r e s i m i l a r ,  ( i n terms of  e s t r o g e n s ) the p r e p u t i a l  glands  i n most r e s p e c t s , t o t h o s e  of t h e  - i i i -  rat  as d e s c r i b e d by o t h e r a u t h o r s .  apparent  d i f f e r e n c e s i n t h e m i c r o s c o p i c anatomy a n d i n t h e  h i s t o c h e m i s t r y of these presence  homologous o r g a n s ;  the l i f e t i m e gland;  granules  of the animal  which p e r s i s t s  and w h i c h i s n o t f o u n d  i n the a c i n a r c e l l s  throughout i n the  o f t h e mouse p r e p u t i a l  gland  i n the a c i n a r c e l l s of the r a t p r e p u t i a l  gland, i n a d d i t i o n to l i p i d  the  preputial  s e c o n d l y , t h e absence of p e r i n u c l e a r p r o t e i n a c e o u s  which are present  in  f i r s t l y , the  o f an i n t i m a t e a s s o c i a t i o n o f t h e mouse  gland with a central h a i r f o l l i c l e  rat  However, t h e r e a r e  droplets.  Lipids,  primarily  t h e f o r m o f d r o p l e t s o f n e u t r a l f a t s a r e , t o my k n o w l e d g e , only secretory products Developmentally,  o f t h e mouse p r e p u t i a l  gland.  h i s t o l o g i c a l l y , and h i s t o c h e m i c a l l y ,  t h e p r e p u t i a l g l a n d o f t h e mouse i s a n e x c e s s i v e l y d e v e l o p e d s e b a c e o u s g l a n d and u n l i k e t h e r a t p r e p u t i a l g l a n d c a n n o t be c a t e g o r i z e d so s i m p l y , b e l o n g s v a r i e t y of h o l o c r i n e gland.  which  t o the "monocrine"  -ivTABLE OF CONTENTS Page T i t l e Page  .  i  Abstract  i i  Table of Contents  iv  List  of Tables  v  List  of Graphs  v i  List  of P l a t e s  v i i  Introduction  1  M a t e r i a l s and Methods  3  Results  11  G r o s s Anatomy Microscopic  of the A d u l t P r e p u t i a l Gland  Anatomy  of the A d u l t  D e v e l o p m e n t and A g i n g  ..  Gland  11 16  .  20  Physiology  27  Histochemistry  48  Discussion  54  Histology  54  Physiology  59  Histochemistry  75  Summary and C o n c l u s i o n s  78  Illustrations  82  Acknowledgements  101  Literature Cited  102  -V-  L I S T OF TABLES  TABLE  I II  III  IV  PAGE  Summary o f E x p e r i m e n t a l P r o c e d u r e f o r the P h y s i o l o g i c a l Study  . 6  Normal V a r i a t i o n s i n Body W e i g h t and W e i g h t s o f S i n g l e P r e p u t i a l G l a n d s i n W e a n l i n g and Young A d u l t Mice  1 5  Average R e l a t i v e W e i g h t s o f S i n g l e P r e p u t i a l Glands i n the P h y s i o l o g i c a l Study  29  Computer Program A n a l y s i s P h y s i o l o g i c a l Study  32  of the  -vi-  L I S T OF GRAPHS  GRAPH  1  2  PAGE  Body W e i g h t s . a n d W e i g h t s o f S i n g l e P r e p u t i a l G l a n d s o f Male M i c e From 1 Week t o 12 Months  13  Body W e i g h t s and W e i g h t s o f S i n g l e P r e p u t i a l G l a n d s o f Female M i c e From 1 Week t o 12 Months  14  -vii-  L I S T OF PLATES  PLATE  PAGE  1  The Young A d u l t P r e p u t i a l  Gland  82  2  The Young A d u l t P r e p u t i a l  Gland  83  3  The D e v e l o p m e n t o f t h e P r e p u t i a l  G l a n d ..  84  4  The D e v e l o p m e n t o f t h e P r e p u t i a l  Gland . .  85  5  The A g i n g of t h e P r e p u t i a l  Gland  86  6  The A g i n g o f t h e P r e p u t i a l  Gland  87  7  The E x p e r i m e n t a l G l a n d s o f Group I (Males)  88  The E x p e r i m e n t a l G l a n d s o f Group I (Males)  89  The E x p e r i m e n t a l G l a n d s o f Group I ' (Females )  90  .8 9 10  The E x p e r i m e n t a l G l a n d s o f G r o u p I (Females)  91  11  The E x p e r i m e n t a l G l a n d s o f G r o u p I i I n t a c t A d u l t Mice (Males)  92  The E x p e r i m e n t a l G l a n d s o f Group I i I n t a c t A d u l t Mice (Females)  93  13  The E x p e r i m e n t a l G l a n d s o f Group I I V e a n l i n g Mice (Males)  94  14  The E x p e r i m e n t a l G l a n d s o f Group I I Weanling Mice (Females)  95  15  The E x p e r i m e n t a l G l a n d s o f G r o u p Newborn M i c e  Ill96  The E x p e r i m e n t a l G l a n d s o f G r o u p Newborn M i c e  Ill-  12  16  97  -viii-  PLATE  PAGE  17  Histochemistry  98  18  Histochemistry  99  19  Histochemistry  100  -1-  INTRODUCTION  The p r e p u t i a l g l a n d s are p a i r e d subcutaneous sexes, side  musculus,  organs which are s i t u a t e d , i n both  i n t h e g e n i t a l r e g i o n o f t h e a n i m a l , one on e i t h e r  of the m i d l i n e .  the s k i n , b e s i d e  E a c h g l a n d opens  and a l o n g t h e l a t e r a l  female  independently  onto  the u r e t h r a l o p e n i n g , v i a an e x c r e t o r y d u c t  which runs along the l a t e r a l mouse  o f t h e mouse, Mus  side  of the p e n i s  i n t h e male  s i d e of the c l i t o r i s i n the  mouse. U n l i k e the r a t p r e p u t i a l glands  w h i c h have b e e n t h e  c o n c e r n of d e t a i l e d h i s t o l o g i c a l , h i s t o c h e m i c a l , e n d o c r i n o logical  and b i o c h e m i c a l i n v e s t i g a t i o n s  the p r e p u t i a l glands little  attention.  Although Voss  of the g l a n d s  i n v o l v i n g the response  of the g l a n d to androgens  has  preputial  (Voss,  1932;  and more r e c e n t l y , i n f o r m a t i o n on (Bronson,  1967).  My s t u d y r e p o r t s t h e o b s e r v a t i o n s histology  adult  investigations  sex a t t r a c t a n t f u n c t i o n f o r t h e g l a n d  Gaunt,  (1933)  t o my k n o w l e d g e ,  endocrinological  B u r d i c k and Gamon, 1941)  1966;  i n the normal  O t h e r r e f e r e n c e s t o t h e mouse  gland concern cursory  a possible  ( 1 9 3 2 ) and S c h a f f e r  t h e i r d e t a i l e d morphology,  not been d e s c r i b e d .  authors,  of t h e mouse have r e c e i v e d c o m p a r a t i v e l y  give b r i e f d e s c r i p t i o n s male mouse,  b y a number o f  of t h e p r e p u t i a l g l a n d s  p o s t - n a t a l d e v e l o p m e n t and a g i n g  made upon t h e  of normal mice and a l s o  during  the h i s t o l o g y  of  -2-  the glands  of e x p e r i m e n t a l a n i m a l s , a f t e r hormone  i n order to determine  t h e i r p o s s i b l e endocrine  treatments,  relationships.  F i n a l l y , r o u t i n e h i s t o c h e m i c a l t e s t s f o r l i p i d s and p r o t e i n s were c a r r i e d  o u t on e x c i s e d p r e p u t i a l g l a n d s  i d e n t i f y and  to l o c a l i z e  p a p e r of B e a v e r proved  their  secretory products.  The  ( i 9 6 0 ) on t h e p r e p u t i a l g l a n d s o f t h e r a t  a v a l u a b l e p o i n t o f r e f e r e n c e f o r my  mouse p r e p u t i a l  i n order to  gland.  s t u d y on  the  -3-  MATERIALS AND METHODS  Histology For the h i s t o l o g i c a l subsequent  aging  study  of t h e d e v e l o p m e n t  and  of the p r e p u t i a l g l a n d , w h i t e Swiss m i c e ,  b o t h male and f e m a l e , o f d i f f e r e n t ages r a n g i n g  from  t o 12 months  glands  o l d were s a c r i f i c e d and t h e p a i r e d  removed.  In mice, from b i r t h u n t i l  necessary  to d i s s e c t  7 days of age,  the whole g e n i t a l a r e a .  newborn  it  Animals  was were  s a c r i f i c e d a t w e e k l y i n t e r v a l s f r o m 7 d a y s o f age u n t i l weeks months  o f age and t h e r e a f t e r a t m o n t h l y i n t e r v a l s u n t i l of a g e .  e x c e p t i o n of newborn,  1,  2, 3, 4 ,  W i t h the  5, and 6 day o l d  glands  w h i c h c o u l d n o t be i d e n t i f i e d g r o s s l y and t h e r e f o r e a l l g l a n d s were t r e a t e d s i m i l a r l y  fixation,  p r i o r to weighing.  of excess  periglandular  on a S a r t o r i u s  The g l a n d s were d i s s e c t e d  f a t , b l o t t e d dry,  balance.  and t h e n  free  weighed  F i n a l l y , a l l t h e g l a n d s were  D e l a f i e l d ' s h e m a t o x y l i n and c o u n t e r s t a i n e d w i t h Hormone  could  after  i n p a r a f f i n , s e c t i o n e d s e r i a l l y a t 8 p.. and s t a i n e d  Physiology:  12  The d i s s e c t e d g l a n d s were f i x e d i m m e d i a t e l y  i n B o u i n ' s f i x a t i v e f o r a p p r o x i m a t e l y 24 h o u r s .  n o t be w e i g h e d ,  8  embedded  with  eosin.  Treatments  For the p h y s i o l o g i c a l  s t u d y w h i t e mice of the Swiss •  s t r a i n were d i v i d e d i n t o f o u r m a j o r g r o u p s a c c o r d i n g t o  age  and s t a t u s .  7  weeks)  Group I  consisted  which at the onset  o f y o u n g a d u l t mice  of t h e e x p e r i m e n t were  (age  castrated  _4-  (by s u r g i c a l incision)  r e m o v a l of t h e gonads t h r o u g h  an  abdominal  and had t h e l e f t p r e p u t i a l g l a n d removed a t  same t i m e .  For t h i s  o p e r a t i o n t h e m i c e were  w i t h an i n t r a p e r i t o n e a l i n j e c t i o n o f A v e r t i n  the  anaesthetized solution.  ( 0 . 0 2 5 m l . of A v e r t i n p e r one gram o f body w e i g h t was adequate stock  dosage.  solution:  ethanol,  1.0  The A v e r t i n was p r e p a r e d f r o m t h e 1.0  m l . amylene h y d r a t e ,  ml. d i s t i l l e d water.  was made up t o 20 m l . o f of d i s t i l l e d w a t e r . ) (age  7 weeks)  the l e f t  mice  (age  0.5 m l . t r i b r o m o -  Then 0.5 m l . o f t h e  consisted  which a t the b e g i n n i n g  at the onset  Group I I  of t h e e x p e r i m e n t  of the e x p e r i m e n t .  was made up o f  f o u r major  was t h e n d i v i d e d i n t o s u b g r o u p s a , b , £ , to the treatment g i v e n .  r e c e i v e d a d a i l y subcutaneous testosterone  propionate*  and d a c c o r d i n g  i n j e c t i o n o f 0.125  25mg/cm , C i b a Company L i m i t e d ) .  i n j e c t i o n o f 0.00165 mg.  Biochemicals  In  i n j e c t e d d a i l y w i t h 0.05  (*Perandren  E a c h mouse i n s u b g r o u p b  m l . o f sesame o i l  Corporation) .  and  mg.of  m l . o f sesame o i l  e s t r a d i o l b e n z o a t e * i n 0.05  i n s u b g r o u p d were n o t  groups  E a c h mouse i n s u b g r o u p a  i n 0.05  r e c e i v e d a d a i l y subcutaneous  removed  consisted  There w e r e , o f c o u r s e , male  female mice i n each g r o u p i n g .  had  weanling  F i n a l l y , Group I I I  Each of t h e s e  ml.  anaesthesia  w h i c h had t h e l e f t p r e p u t i a l g l a n d  o f i n t a c t newborn m i c e .  stock  of young a d u l t mice  p r e p u t i a l g l a n d removed u n d e r A v e r t i n  3 weeks)  following  s o l u t i o n b y t h e a d d i t i o n o f 19.5  Group I i  b u t were n o t c a s t r a t e d .  an  of  (^Nutritional  s u b g r o u p c_ e a c h a n i m a l  m l . of sesame o i l U . S . P .  was  The m i c e  i n j e c t e d or h a n d l e d a f t e r the  original  -5-  o p e r a t i o n , but served as c o n t r o l s f o r the other subgroups  within  the same major group.  A l l groups I , I i , and I I I animals were  kept on the experiment  f o r 21 days.  however, were on the experiment  Group II animals,  f o r 19 days only.  summarizes the above experimental procedure.  Table I  At the end of  each experiment,24 hours a f t e r the l a s t i n j e c t i o n , a l l the mice were s a c r i f i c e d and the remaining p r e p u t i a l gland, t h a t i s the r i g h t gland, was removed from each animal i n groups I, I i , and I I and both p r e p u t i a l glands were removed from each animal i n group I I I .  As soon as the p r e p u t i a l  glands  were d i s s e c t e d , e i t h e r at the onset of each experiment the end of each experiment,  or at  they were t r e a t e d s i m i l a r l y .  p r e v i o u s l y d e s c r i b e d f o r the h i s t o l o g i c a l  study, the glands  were f i x e d i n Bouin's f i x a t i v e and t h e r e a f t e r d i s s e c t e d of excess f a t , b l o t t e d dry, and weighed.  As  free  (The p a i r e d glands  from each mouse of group I I I , the newborn group, were weighed together and then averaged; the two glands from each animal of the a d u l t and weanling groups  ( I , I i , II)  were weighed s i n g l y so that the weights before and a f t e r treatment could be compared.)  F i n a l l y , the glands were  embedded i n p a r a f f i n , s e r i a l l y s e c t i o n e d a t 8 ji.  and s t a i n e d  with hematoxylin and e o s i n . Histochemistry Routine h i s t o c h e m i c a l t e s t s f o r l i p i d s and p r o t e i n s were c a r r i e d out on the e x c i s e d p r e p u t i a l glands of young a d u l t Swiss mice (5-8 weeks) and of weanling mice, of both  ;  TABLE I SUMMARY OF EXPERIMENTAL PROCEDURE FOR THE PHYSIOLOGICAL STUDY  Group  T  Ii  Treatment Subgroup  Sex a n d No. o f Animals  Daily Dosage/ Mouse  Total Hormone Injected/ Mouse  Duration Age a n d s t a t u s of a t onset of Experiment Experiment  a)Testosterone propionate  M 5 F 5  0.125 mg  2. 625 mg  21 d a y s  b)Estradiol benzoate  M 5 F 3  0.00165 mg  0 . 03465 mg  21 d a y s  c)Sesame O i l U.S.P.  M 5 F 5  0.05 ml  d)Control  M 2 F 4  a)Testosterone propionate  M 3 F 3  0.125 mg  2. 625 mg  21 d a y s  b)Estradiol benzoate  M 3 F 2  0.00165 mg  0 . 03465 mg  21 d a y s  c)Sesame O i l U.S.P.  M 1 F 1  0.05 m l  d)Control  M 7 F 6  21 d a y s  Young a d u l t m i c e (age 7 "weeks) , c a s t r a t e d and left preputial g l a n d removed simultaneously at beginning of experiment  21 d a y s  21 d a y s 21 d a y s  I n t a c t , young a d u l t m i c e (age 7 weeks). Left preputial gland removed a t beginning of experiment  TABLE I c o n t .  Group  II  III  Sex and No. of Animals  DailyDosage/ Mouse  Total Hormone Injected/ Mouse  Duration of Experiment  a)Testosterone propionate  M 3 P 5  0.125 mg  2.375 mg  19 d a y s  b)Estradiol benzoate  M 4 F 5  0.00165 mg  0.03135 mg  19 d a y s  c)Sesame O i l U.S.P.  M 5 F 6  0.05 m l  d)Control  M 5 F 6  a)Testosterone propionate  M 4 F 6  0.125 mg  2.625 mg  21 d a y s  b)Estradiol benzoate  M 3 F 4  0.00165 mg  0.03465 mg  21 d a y s  c)Sesame O i l U.S.P.  M 4 F 5  0.05 ml  Treatment Subgroup  Age a n d s t a t u s a t onset of Experiment Intact veanling m i c e (age 3 w e e k s ) Left preputial g l a n d removed a t onset o f experimen  -  19 d a y s 19 d a y s  21 d a y s  Intact mice  newborn  -8-  sexes,  i n o r d e r t o l o c a l i z e and t o i d e n t i f y , a t l e a s t ,  to class,  the s e c r e t o r y p r o d u c t s  as  o f the a c i n a r c e l l s .  All  g l a n d s f o r h i s t o c h e m i c a l s t u d i e s were f i x e d i n 10%> b u f f e r e d neutral formalin, unless, The b u f f e r was 12-24  otherwise,  sodium p h o s p h a t e .  i n d i c a t e d by the method.  F i x a t i o n time v a r i e d  hours depending upon the r e q u i r e m e n t  of the  procedure.  The h i s t o c h e m i c a l and s t a i n i n g methods were c a r r i e d according to Pearse These methods  (1968) e x c e p t as o t h e r w i s e  i n c l u d e d McManus'  compound l i p i d s  Baker's formol calcium f o r 4 weeks);  frozen sections  lipids  of t i s s u e ,  about  out  indicated.  Sudan b l a c k B t e c h n i q u e  i n p a r a f f i n sections  Sudan b l a c k B method f o r  (after fixation the propylene  (Pearse,  in  1961), u s i n g  Butcher,  fresh  the  for  c h o l e s t e r o l which also  the p e r f o r m i c - a c i d S c h i f f u n s a t u r a t e d bonds  requires  the S c h u l t z  fresh, frozen  method f o r l i p i d s  method  sections;  sections);  (Baker's modification);  t h e f e r r i c f e r r i c y a n i d e method f o r s u l p h y d r y l fixed,  70%  containing  (using formalin f i x e d , p a r a f f i n  the M i l l o n r e a c t i o n f o r t y r o s i n e  Sudan  and P o u l t e r  ( 1 9 6 9 ) , u s i n g a s a t u r a t e d s o l u t i o n of Sudan b l a c k B i n a l c o h o l and f r e s h f r o z e n s e c t i o n s ;  for  glycol-  14 ja i n t h i c k n e s s ;  b l a c k B method o f C h a y e n , B i t e n s k y ,  ethyl  from  p a r a f f i n s e c t i o n s and f r e s h , f r o z e n  (using  formalin  sections);  t h e B i e b r i c h s c a r l e t method f o r b a s i c n u c l e o p r o t e i n s  a t pH  level  the  o f 9.5  (following f i x a t i o n i n Carnoy's f l u i d ) ;  B i e b r i c h s c a r l e t method of S p i c e r and L i l l i e staining  60 m i n u t e s  (Lillie,  i n 0.01% B i e b r i c h s c a r l e t i n the  1965) Mcllvaine  -9citric  acid-sodium phosphate  b u f f e r a t pH 4.95; Heidenhain's  i r o n hematoxylin procedure; and M a l l o r y ' s phosphotungstic a c i d hematoxylin method ( C u l l i n g , 1957).  The l a t t e r three  methods, the B i e b r i c h s c a r l e t method of S p i c e r and L i l l i e , Heidenhain's  i r o n hematoxylin method, and M a l l o r y ' s P.T.A.H.  method were a l s o c a r r i e d out on the p r e p u t i a l glands of a d u l t female a l b i n o r a t s for  (200 gm.), f o l l o w i n g f o r m a l i n f i x a t i o n  12 hours, i n order to compare the f i n d i n g s of these  methods on the r a t gland with those on the mouse gland. a d d i t i o n to l i p i d d r o p l e t s , S c h a f f e r (1933) and Beaver  In (1959,  i960) have d e s c r i b e d numerous cytoplasmic granules i n the a c i n a r c e l l s of the r a t p r e p u t i a l gland which are quite d i s t i n c t from the f a t d r o p l e t s and are best  demonstrated  w i t h i r o n hematoxylin or with M a l l o r y ' s s t a i n .  However, the  demonstration of the granules, even with these methods, i s v e r y much dependent upon proper t i s s u e f i x a t i o n ; 10% b u f f e r e d n e u t r a l f o r m a l i n renders the granules i n s o l u b l e so that they r e s i s t almost every subsequent  treatment whereas a l c o h o l i c  and c e r t a i n a c i d f i x a t i v e s d e s t r o y the granules (Beaver, In  order to ensure the exact d u p l i c a t i o n of Beaver's  technique, the B i e b r i c h s c a r l e t method, Heidenhain's  1959).  (i960) iron  hematoxylin method and M a l l o r y ' s P.T.A.H. method were a l s o c a r r i e d out on r a t and mouse glands which had been f i x e d i n 10% n e u t r a l f o r m a l i n b u f f e r e d by sodium acetate  (Lillie,  1965), as w e l l as on those glands f i x e d i n sodium buffered formalin. which Beaver  F i n a l l y , the d i f f e r e n t i a l  phosphate  s t a i n i n g method  (i960) devised to e x h i b i t the s e c r e t i o n granules  -10-  and f a t d r o p l e t s simultaneously i n f r o z e n s e c t i o n s of r a t p r e p u t i a l gland was  c a r r i e d out on the p r e p u t i a l glands of  a d u l t male and female mice and a l s o on the glands of male and female r a t s  (200 gm.).  simultaneous d i f f e r e n t i a l  A second method f o r the s t a i n i n g of both s e c r e t o r y products  i n the r a t p r e p u t i a l gland was  devised u s i n g o i l red 0 and  i r o n hematoxylin (Heidenhain's method): 1)  F i x t i s s u e i n 10% b u f f e r e d n e u t r a l f o r m a l i n f o r 12 hours. (The b u f f e r was sodium phosphate).  2)  Cut t h i n f r o z e n s e c t i o n s about 7u i n t h i c k n e s s .  3)  S t a i n f o r 10 minutes w i t h o i l red 0 i n i s o p r o p a n o l d i l u t e the stock s o l u t i o n of o i l red 0 (lgm. o i l red 0: 500 ml. 99% isopropanol) as f o l l o w s before u s i n g : 9 ml. stock: 6 ml. d i s t i l l e d water, adding the water to the o i l red 0. Let stand 5-10 minutes, then f i l t e r twice. (These three steps f o l l o w the method of Beaver, 1960.)  4)  Rinse i n d i s t i l l e d  5)  Mordant i n a 4%> aqueous s o l u t i o n of ammonium i r o n alum at 45 C f o r 60 minutes.  6)  Rinse i n tap water.  7)  Rinse i n d i s t i l l e d  8)  S t a i n i n 0.5% aqueous s o l u t i o n of hematoxylin at 45 C f o r 60 mintites.  9)  Rinse, i n tap water.  10)  Differentiate i r o n alum.  11)  Blue i n tap water.  12)  Mount i n Apathy's RESULTS:  water.  water.  i n 2.5% aqueous s o l u t i o n of ammonium  medium.  f a t red; granules b l a c k .  -11-  RESULTS  Gross Anatomy of the Adult P r e p u t i a l Gland The p a i r e d p r e p u t i a l glands of the mouse are s i t u a t e d subcutaneously on each side of the m i d l i n e i n the g e n i t a l r e g i o n of the animal.  The e x c r e t o r y duct of each gland  runs along the l a t e r a l  side of the penis i n the male and along  the  lateral  side of the c l i t o r i s i n the female to open  independently beside the u r e t h r a l opening.  A network of  blood v e s s e l s may be seen upon the d o r s a l surface of each gland. In the male mouse, the gland i s a w e l l developed, f l a s k - s h a p e d s t r u c t u r e ; i t ranges i n length from 3 to 10  mm.,  and i n wet weight from 8 to 81 mg., depending on the age of the  a d u l t animal (Graph l ) .  In the a d u l t female mouse, the  p r e p u t i a l gland i s a r a t h e r small, round s t r u c t u r e ; i t averages 2 mm.  i n l e n g t h and ranges from 2 to 7 mg. i n wet  weight, depending a l s o on the age of the animal (Graph 2 ) . There i s considerable v a r i a t i o n i n the weight of p r e p u t i a l glands even among mice of the same age which i s p a r t i a l l y r e l a t e d to a s i m i l a r v a r i a t i o n i n body weight among such animals (Table I I ) . C o n t r a l a t e r a l glands i n the same male animal are not always comparable by weight e i t h e r , there being as much as 10 mg. d i f f e r e n c e .  NOTES TO GRAPHS 1 AND  2  Each p o i n t on the graph r e p r e s e n t s the body weight (gm.) or s i n g l e p r e p u t i a l weight (mg.) of a d i f f e r e n t mouse. ¥here more than one animal of the same age was used, the graph passes through the average weight recorded f o r that age. At 3 weeks, the minimum (min.), maximum (max.) and average (av.) s i n g l e p r e p u t i a l gland weights i n d i c a t e d on graph 1 are the v a l u e s compiled from a t o t a l of 17 weanling males i n experiment I I . At 7 weeks, the min., max., and av. are the v a l u e s compiled from a t o t a l of 31 a d u l t males i n experiments I and I i . At 3 weeks, the min., max., and av. s i n g l e p r e p u t i a l weights i n d i c a t e d on graph 2 are the v a l u e s compiled from a t o t a l of 22 weanling females i n experiment I I . At 7 weeks, the min.;, max., and av. are the v a l u e s compiled from a t o t a l of 29 a d u l t females i n experiments I and I i .  -13GRAPH 1  a b  BODY WEIGHTS AND WEIGHTS OF SINGLE GLANDS OF MALE MICE FROM 1 WEEK TO 12 MONTHS LEGEND  1 2 34 56 78 WEEKS  3  4  5 6 MONTHS  7  8  9  10  11  -14-  GRAPH 2ac BODY WEIGHTS AND WEIGHTS OP SINGLE PREPUTIAL GLANDS OP FEMALE MICE FROM 1 WEEK TO 12 MONTHS  LEGEND 'Body Weight ( gm. ) -•Single P r e p u t i a l Weight (mg.) 40  30  A-  20  10  AL  iin  I I I ' I ' I  '  12 34 5678 WEEKS  3  5  6 7 MONTHS  8  9  10  11  12  TABLE I I Normal v a r i a t i o n s i n body w e i g h t and w e i g h t s o f s i n g l e p r e p u t i a l g l a n d s * i n w e a n l i n g and young a d u l t mice (^compiled from w e i g h t s of p r e p u t i a l g l a n d s removed from mice o f e x p e r i m e n t s I , I i , and I I a t o n s e t o f e a c h experiment). BODY WEIGHTS (gm.) Status  Minimum  No. 31  18.5  27.5  23.7  Adult F  29  16.2  25.1  21.0  Weanling M  17  6.2  13.2  9.2  Weanling F  22  6.5  11.6  9.1  No.  Minimum  Adult M  31  21.0  Adult F  29  0.6  Weanling M  17  F  22  Weanling  Average  Adult M  ABSOLUTE PREPUTIAL WEIGHT (mg.) Status  Maximum  Maximum  Average  RELATIVE PREPUTIAL WEIGHT (mg/lOgm body wt) Minimum Maximum Average  33.0  9.7  20.3  3.4  2.1  0.4  1.9  1.0  1.0  3.5  1.8  1.4  3.0  1.9  0.4  1.5  0.8  0.5  1.4  1.0  49.3  13.8  -16-  M i c r o s c o p i c Anatomy of the Young A d u l t P r e p u t i a l At 5-6 weeks, the age mice of the Swiss s t r a i n , f u l l y developed pattern.  of sexual m a t u r i t y of most  the p r e p u t i a l gland e x h i b i t s a  h o l o c r i n e , compound a l v e o l a r  structural  In both the male and female mouse the parenchyma  of the gland c o n s i s t s of densely packed a c i n i s i z e s and  Gland  i n v a r i o u s s t a t e s of a c t i v i t y ,  of v a r i o u s  a number of which  can be d e s c r i b e d i n the manner of Montagna and Noback (1946) as f o l l o w s : 1)  small, e p i t h e l i o i d  parenchymal a c i n i ; have l i t t l e 2)  a c i n i between the  the c e l l s of these  larger  "primordial a c i n i "  cytoplasm.  rounded a c i n i , more uniform i n s i z e  and  l a y e r of f l a t t e n e d p e r i p h e r a l c e l l s surrounds  shape; a c e l l s of  i n c r e a s i n g magnitude toward the centre of each acinus; these c e l l s have v a c u o l a t e d or granular cytoplasm 3)  g r e a t l y enlarged a c i n i  vacuolated c e l l s  4)  composed of hypertrophied,  (the vacuoles represent cytoplasmic  g l o b u l e s ) surrounded cells  ( F i g . 3).  fatty  by a l a y e r of f l a t t e n e d p e r i p h e r a l  (Fig. 4). acini  a l s o composed of h y p e r t r o p h i e d , v a c u o l a t e d  cells  which are beginning to degenerate i n the centre of the a c i n u s ; these show degenerating (Fig. 5) way;  c e l l membranes  and pyknotic  nuclei  5). acini  i n which c e l l u l a r  a distinct  degeneration  i s w e l l under  lumen appears i n the acinus; o f t e n at  this  -17-  stage  s e v e r a l a c i n i have f u s e d t o f o r m g i a n t  of d i s i n t e g r a t i n g  cells.  The s m a l l e r a c i n i  (types  1, 2 a b o v e )  i n the p e r i p h e r y of the p r e p u t i a l g l a n d . be s e e n i n t h e b a s a l  c e l l s of these  p o s s i b i l i t y of f u r t h e r i n c r e a s e large  acini,  undergoing  ation  (types  3,4,  the  conglomerates  a r e most  numerous  Often mitoses  can  a c i n i i n d i c a t i n g the  i n t h e i r c e l l number.  h y p e r t r o p h y and s u b s e q u e n t  5) a r e most numerous  The  degener-  i n the c e n t r e  of  gland. The above  states  of a c t i v i t y r e p r e s e n t the c y c l e  of  sebum f o r m a t i o n i n t h e p r e p u t i a l g l a n d , f r o m t h e f a t - f r e e acinus  to the h y p e r t r o p h i e d , f a t - l a d e n a c i n u s  degeneration is (type l )  r e p l a c e those  Each a c i n u s fatty  increases  substances  central  taking place.  i n which  The s m a l l p r i m o r d i a l a c i n i  a c i n i w h i c h have a l r e a d y f o r m e d  sebum.  i n s i z e w i t h the c o n t i n u e d storage  of  i n the a c i n a r c e l l s .  c e l l s begin to degenerate.  p y k n o t i c , and c e l l u l a r membranes  Then, the  hypertrophied  The n u c l e i become  indistinct.  Cellular  breakdown p r o g r e s s e s from the c e n t r e of the a c i n u s the p e r i p h e r y .  toward  The f l a t t e n e d p e r i p h e r a l c e l l s o f t h e  m e a n w h i l e , become s t r a t i f i e d t o f o r m t h e e p i t h e l i a l (2-3  layers thick)  which,  cell  o f a new a c c e s s o r y  duct i n the  acinus,  lining  gland,  i n o r d e r t o open i n t o a d u c t a l r e a d y p r e s e n t  i n the  g l a n d , must b r e a k t h r o u g h t h e s t r a t i f i e d e p i t h e l i u m o f old duct.  The edges o f t h e e p i t h e l i u m a t t h i s  site  the  become  f r a y e d and a r e c a s t o f f i n t o t h e sebum.  Eventually  the  lining  continuous  with  o f t h e new a c c e s s o r y  d u c t becomes  -18-  t h a t of the o l d duct, p e r m i t t i n g the passage of the sebum from the degenerated The  acinus to the c e n t r a l duct of the gland.  complete sequence of a c t i v i t y i n the c y c l e of  sebum formation i s not l i m i t e d to the uniform types of a c i n i just described.  I t i s seen, too, i n the l a r g e  conglomerates  of a c i n i i n which there i s a gradual accumulation  of  lipid  d r o p l e t s i n the a c i n a r c e l l s , p r o g r e s s i n g from the p e r i p h e r y to the centre of the a c i n i , near a duct, where c e l l u l a r degeneration, subsequently, takes place ( F i g . 6). The  c y c l e of h o l o c r i n e s e c r e t i o n and duct formation,  which has been d e s c r i b e d above, has been termed " d u c t u l a r t r a n s f o r m a t i o n " by Beaver (i960).  This seems to be a u s e f u l  d e s c r i p t i v e term to denote a c t i v e s e c r e t i o n and w i l l t h e r e f o r e be used throughout  t h i s paper to r e f e r to the a c i n a r c y c l e .  The p a t t e r n of ducts i n the p r e p u t i a l gland i s always changing The  as new  l a t e r a l or accessory ducts are formed.  l a t e r a l ducts open i n t o the c e n t r a l duct, which i n the  5 week o l d mouse i s most extensive i n the centre of the gland.  The  epithelium  c e n t r a l duct i s l i n e d by s t r a t i f i e d ,  squamous  (4-6 l a y e r s t h i c k ) which i s not k e r a t i n i z e d .  It,  i n t u r n , connects w i t h the main e x c r e t o r y duct of the gland which i s l i n e d by s t r a t i f i e d  squamous e p i t h e l i u m (about  8-10  l a y e r s t h i c k ) continuous w i t h that of the epidermis of the surrounding g e n i t a l r e g i o n .  The  squamous e p i t h e l i u m of the  main e x c r e t o r y duct i s f o r the most p a r t k e r a t i n i z e d  (like  the e p i d e r m i s ) , and thus k e r a t o h y a l i n e granules, s t a i n e d  -19-  heavily layers  b y h e m a t o x y l i n , c a n be s e e n i n t h e s u p e r f i c i a l of f l a t t e n e d e p i t h e l i a l  excretory with  cells  duct i s not k e r a t i n i z e d  the c e n t r a l  ( P i g . 15).  The m a i n  d i s t a l l y , near i t s union  duct.  The p r e p u t i a l g l a n d i s s u r r o u n d e d b y a c o n s p i c u o u s connective tissue  c a p s u l e w h i c h i s composed  of c o l l a g e n o u s  f i b r e s , f i b r o b l a s t s , m a c r o p h a g e s and a d i p o s e t i s s u e . continuous with within  Itis  the c o n n e c t i v e t i s s u e network or stroma  the gland i t s e l f , which d e l i c a t e l y surrounds  a c i n u s and u n d e r l i e s  the epithelium  each  of each duct.  Occasionally,  t h i c k e r masses o f i n t e r s t i t i a l  connective  tissue  the parenchyma i n t o i n d e f i n i t e  lobes.  divide  interstitial numerous  tissue  i s also  richly vascularized;  there are  capillaries.  I n b o t h t h e male and f e m a l e g l a n d a s i n g l e follicle  i s intimately associated  stitially. sections section the  the  The h a i r f o l l i c l e  and i t s s t r u c t u r e  inner  root  distinguished  i s like  An o u t e r r o o t  inter-  through  either  near the d i s t a l  that  serial  i n crossend o f  projects  o f any h a i r f o l l i c l e o f  sheath surrounds the c e l l s  sheath which, i n t u r n ,  The h a i r s h a f t  the gland,  hair  d u c t surrounded by the parenchyma of the  The s t r u c t u r e  skin.  with  c a n be f o l l o w e d  or l o n g i t u d i n a l s e c t i o n ,  main e x c r e t o r y  gland.  The  surrounds the h a i r  i n t o the p o s t e r i o r  duct near i t s union w i t h  the main e x c r e t o r y  of the shaft.  end o f t h e c e n t r a l duct ( F i g . 7 ) .  -20-  B e s i d e s the obvious weight  ( G r a p h s 1,  gross d i f f e r e n c e s i n size  and  2) t h e r e a r e c e r t a i n s e x d i f f e r e n c e s i n  t h e s t r u c t u r a l p a t t e r n o f t h e p r e p u t i a l g l a n d s o f 5 week male and  female  mice.  I n the male, the d u c t system  has  become e x t e n s i v e by d u c t u l a r t r a n s f o r m a t i o n ( P i g . l ) . Conversely, developed  i n the female  ( P i g . 2).  fibrous i n t e r s t i t i a l  the duct system  i s less  Furthermore,  i n the female,  t i s s u e which  surrounds  well the  the a c i n i i s  f r e q u e n t l y t h i c k e r t h a n i n t h e male c o u n t e r p a r t ( F i g . 8).  D e v e l o p m e n t and  Aging  I n t h e newborn male and  female  mouse e a c h p r e p u t i a l  g l a n d c o n s i s t s of s e v e r a l masses o f p r i m o r d i a l a c i n a r c e l l s i n the  s u r r o u n d i n g c o n n e c t i v e t i s s u e of the  ( F i g . 9). tissue  At t h i s  capsule.  prepuce  s t a g e t h e r e i s no d e f i n i t e  There a r e m i t o t i c  of the a c i n a r c e l l s .  Other  cells  connective  configurations i n a i n the  c e n t r e of the  a r e l a d e n w i t h f a t t y d r o p l e t s and have b e g u n t o by a p r o c e s s  s i m i l a r to t h a t which  of the a d u l t g l a n d .  The  ( F i g . 10).  epidermis a c t u a l l y gives r i s e t h a t the s t r a t i f i e d  degenerate  takes p l a c e i n the  ( F i g . 10).  acini  An  the d e g e n e r a t i o n of i n v a g i n a t i o n of  to the main e x c r e t o r y duct  The  c o n t i n u i t y w i t h the  lumen of t h e e x t r e m e  p o r t i o n of the duct r e s u l t s from  the these  the  squamous e p i t h e l i u m o f t h e d u c t  is also k e r a t i n i z e d ) maintains epithelium  gland  lumen of t h e d i s t a l p o r t i o n o f  main e x c r e t o r y duct i s d e r i v e d from hypertrophied c e l l s  few  so  (which surface  proximal  the desquamation or  -21-  sloughing o f f of e p i t h e l i a l  cells  l a y e r of cuboidal c e l l s which the c u b o i d a l c e l l s surrounds  at this  site.  A  single  seems t o be c o n t i n u o u s w i t h  of the e p i t h e l i u m of the e x c r e t o r y duct  the p r i m o r d i a l a c i n a r c e l l s .  At b i r t h  the intimate  a s s o c i a t i o n between t h e p r e p u t i a l g l a n d and a h a i r is present  ( F i g . 11).  primordial acinar cells distal in  follicle  l i e s next to the  a n d t h e h a i r s h a f t emerges i n t o t h e  end o f t h e m a i n e x c r e t o r y d u c t  the a d u l t gland.  cells  The h a i r f o l l i c l e  ( F i g . 1 2 ) as i t does  The o u t e r r o o t s h e a t h c e l l s  of the h a i r  are continuous w i t h the cuboidal-shaped,  of the gland.  follicle  peripheral  This i s not s u r p r i s i n g since a l l these  s t r u c t u r e s are d e r i v a t i v e s  of the s k i n .  D u r i n g t h e f i r s t week o f d e v e l o p m e n t o f t h e p r e p u t i a l g l a n d , i n t h e male and f e m a l e mouse a f t e r b i r t h , gradual increase i n size,  as t h e r e s u l t o f t h e  there i s a proliferation,  by m i t o s i s , o f t h e p r i m o r d i a l a c i n a r c e l l s  of the gland.  t h e end o f t h e f i r s t week, t h e g l a n d s a r e  definitely  identifiable  structures grossly.  z a t i o n of the gland toward begins.  Microscopically, reorgani-  the a d u l t s t r u c t u r a l p a t t e r n  I n o t h e r w o r d s , some o f t h e a c i n a r c e l l s  t o g e t h e r and v a g u e l y r e s e m b l e adult gland; i r r e g u l a r l y  are grouped  t h e type 1 and 2 a c i n i  shaped b a s a l c e l l s  v a t i v e s of the cuboidal-shaped p e r i p h e r a l  Individual  acinar groupings are undergoing  deri-  of the  are i n v a r i o u s  cells within  hypertrophy  of the  (probably  cells  younger gland) surround a c i n a r c e l l s which degrees of enlargement.  By  other  or degeneration  -22-  t o f o r m sebum ( F i g . there  is  13).  Between these  a stroma of f i b r o u s  many f i b r o b l a s t s  and  A t 2 weeks  connective tissue  o f age  the p r e p u t i a l g l a n d i s  a  t h e a c i n i a r e now d e n s e l y p a c k e d t o g e t h e r ,  of the a d u l t g l a n d  (Fig.  14).  compact  more  as t h e s t r u c t u r a l  types  The p e r i p h e r a l c e l l s o f  a c i n i a r e now q u i t e f l a t t e n e d . various  Conglomerates  A stroma of d e l i c a t e c o n n e c t i v e t i s s u e , s u r r o u n d s t h e a c i n i and i s  around the whole  As d e v e l o p m e n t 3 to 5 weeks,  continues  the glands  t h e male g l a n d u n d e r g o e s Microscopically,  intensely  now c o n t i n u o u s  in seen.  eosinophilic,  w i t h the  connective  gland. d u r i n g the i n t e r v a l from  of b o t h s e x e s i n c r e a s e  in size  t h e more s t r i k i n g  change.  size  i n b o t h t h e male and f e m a l e g l a n d ,  3) become more and more n u m e r o u s .  but  the  e n l a r g e d a c i n i composed o f h y p e r t r o p h i e d , v a c u o l a t e d (type  the  of a c i n i  s t a g e s o f d u c t u l a r t r a n s f o r m a t i o n c a n a l s o be  capsule  of  I n b o t h t h e male and f e m a l e  u n i f o r m l y shaped, and r e c o g n i z a b l e  tissue  composed  macrophages.  structure, microscopically. gland,  "primitive" acini  cells  Ductular  transformation is  extensive  so t h a t e v e n t u a l l y t h e  d u c t of the g l a n d  l i n e d w i t h s t r a t i f i e d squamous e p i t h e l i u m  opens i n t o t h e m a i n e x c r e t o r y d u c t ducts  appear  Finally, (at  i n the substance  (by 3 weeks)  of the g l a n d  and  previously  lateral  (by 4 w e e k s ) .  the g l a n d s a t t a i n the mature h i s t o l o g i c a l  5 t o 6 weeks)  central  d e s c r i b e d and t h e  pattern  sexual  -23-  d i f f e r e n c e s become During parenchyma  apparent.  the i n t e r v a l from 5 to 8 weeks,  i n the gland  o f t h e male mouse  t h e amount  increases  s u b s t a n t i a l l y but the s t r o m a l framework remains  delicate.  A t t h e same t i m e t h e d u c t s y s t e m becomes more e x t e n s i v e d u c t u l a r t r a n s f o r m a t i o n and t h e p a t t e r n o f l a t e r a l is  constantly  week 8,  changed b y t h e f o r m a t i o n o f new o n e s .  By  t h e lumen o f t h e c e n t r a l d u c t , t h e a c i n i a r e m a i n l y  the c o n n e c t i v e t i s s u e of the d u c t s interval  of  i n t h e d u c t s y s t e m and e s p e c i a l l y i n t h e s i z e  to the p e r i p h e r y of the g l a n d ;  (Fig.  capsule  16).  In  from 5 to 8 weeks,  n o t change  greatly  (as  Ductular  the of  confined  a small area  between  and t h e s t r a t i f i e d e p i t h e l i u m  the female g l a n d , d u r i n g  the  t h e amount  does  of parenchyma  i n the male) but the s t r o m a l  becomes more a b u n d a n t : are p r o m i n e n t .  that i s ,  by  ducts  i n the c e n t r a l p o r t i o n of the g l a n d , because  increase  of  fibrous  connective  transformation is  tissue  framework  trabeculae  generally  extensive  and as t h e a c i n i r e g r e s s t o w a r d t h e p e r i p h e r y o f t h e  gland  there  central  is  a compensatory  duct.  Moreover,  gland,  t h e r e a r e few l a t e r a l  i n t h e male  because  increase  of the s m a l l e r ducts  of the p r e p u t i a l g l a n d ,  is  (Fig.  size 17)  of the  female  u n l i k e the  pattern  gland.  The p e r i o d o f m i c r o s c o p i c  secretion  i n t h e lumen of t h e  development  from b i r t h u n t i l  (ductular transformation)  and  organization  8 weeks when a c t i v e  becomes  r e f l e c t e d g r o s s l y by a p e r i o d of s t e a d y  really and  extensive,  pronounced  g r o w t h o f t h e g l a n d by w e i g h t f r o m week 1 t o week 8.  Graphs  -24-  1 and 2 g i v e  the w e i g h t s  ( i n mg.)  for  single  preputial  glands  o f male and f e m a l e m i c e r e s p e c t i v e l y as t h e ages o f  the  animals  graphs  do n o t  change.  It  is  i m p o r t a n t to note t h a t these  show a d e f i n i t i v e g r o w t h c u r v e f o r t h e p r e p u t i a l  g l a n d of mice but m e r e l y i l l u s t r a t e the g e n e r a l p a t t e r n of the g l a n d female.  i n t h e male and f o r c o m p a r i s o n ,  The i n c r e a s e  considerably  greater  i n t h e male t h a n i n t h e f e m a l e  s t r u c t u r a l p a t t e r n as t h e y u n d e r g o s t r i k i n g age  are of course (Figs.  18 and 1 9 ) ,  increase continues cast  off  months types  (there  has  have a d i f f e r e n t  c e r t a i n age  is  decrease  changes.  in acinar  glands  tissue  cycle)  and t h e  tissue  compensatory  ducts  i n t o t h e sebum t o o .  and v a r i o u s  are g r a d u a l l y  In  destroyed  t h e male g l a n d ,  a compact mass o f a c i n i of  sizes  remains  l o c a t e d i n the  B u t a n t e r i o r l y as t h e  and i n t e r s t i t i a l  tissue  A few l y m p h o c y t e s  numerous posterior  subepithelial  become more a b u n d a n t ,  o f t e n appear  i n the  s t r o m a and some f a t c e l l s a c c u m u l a t e  acinar tissue.  and  from 3  few r e m a i n i n g a c i n i a r e o f t e n i n v a d e d b y c o n n e c t i v e  tissue  The  no r e g e n e r a t i o n of a c i n a r  completed i t s  and t h e l a t e r a l  t o 10 m o n t h s ,  elements.  gland.  i n t h e lumen o f t h e c e n t r a l d u c t as t h e sebum c y c l e  p a r t of the g l a n d . stroma  is  c h a n g e s i n b o t h t h e male and f e m a l e  the continuous  once t h e a c i n u s  i n the  i n w e i g h t f r o m week 1 t o week 8  The p r e p u t i a l g l a n d s o f o l d e r a n i m a l s  most  growth  tissue  connective  i n the  But the e x a c t time of appearance  the  of  interthese  -25-  l a t t e r age  changes depends on the i n d i v i d u a l animal.  c e l l accumulation  i n the i n t e r a c i n a r t i s s u e and  i n v a s i o n of the stroma were observed  Fat  lymphocytic  i n the p r e p u t i a l  glands  of 5, 8, and 9 month o l d males but not i n 6 or 7 month o l d animals which d i d not even have the abundant stroma, c h a r a c t e r i s t i c of aging glands.  In 11 and  12 month o l d male  mice, the p r e p u t i a l glands are c y s t i c ; the c e n t r a l  and  l a t e r a l ducts are f i l l e d with a substance  lymphocytes,  polymorphonuclear epithelial cells  n e u t r o p h i l e s and desquamated duct ( F i g s . 20 and 22).  duct e p i t h e l i u m degenerate lymphocytes.  rich in  Only a few  A c i n i , stroma, and  as they are invaded by numerous  i s o l a t e d a c i n i remain at the  p e r i p h e r y and i n the p o s t e r i o r p o r t i o n of the gland near the main e x c r e t o r y duct.  The  stratified  squamous e p i t h e l i u m  of the main e x c r e t o r y duct does not seem to be a f f e c t e d by the lymphocytes.  In the female  gland at 3 months, the  decrease  i n a c i n a r t i s s u e , accompanied by the i n c r e a s e i n the  size  of the duct system, i s a l r e a d y at an advanced stage.  The  little  remaining parenchyma at the p e r i p h e r y of the gland  around the ducts c o n s i s t s only of small a c i n i ; there are no enlarged, h e a v i l y v a c u o l a t e d a c i n i and no of a c i n i age).  (these are s t i l l  conglomerates  seen i n the male gland at t h i s  During the i n t e r v a l from 3 months to 12 months, the  parenchyma continues to decrease and degenerate;  the lumen  of the c e n t r a l duct becomes l a r g e r as the l a t e r a l ducts are engulfed by i t ;  f a t c e l l s accumulate i n the connective  -26-  tissue  s t r o m a w h i c h becomes more and more a b u n d a n t .  t h e male g l a n d s , cystic  (Fig.  t h e d e g e n e r a t i v e phenomena w h i c h  the weights  w i t h age  of the p r e p u t i a l glands  accompany  do n o t  decrease  i n e i t h e r t h e male o r t h e f e m a l e as G r a p h s  illustrate  f o r each sex r e s p e c t i v e l y .  female gland interval  ( G r a p h 2) r e m a i n s  fairly  f r o m 8 weeks t o 12 months  o f t h e male g l a n d interval,  (Graph l )  The w e i g h t  adult male.  less  and a l t h o u g h t h e  the i n c r e a s e  sates  f o r the l o s s  study  of w e i g h t  the the weight  fluctuates widely during  i n w h i c h age  changes  In  c a n n o t r e v e a l t h e changes  n o r m a l l y i n the p r e p u t i a l glands a h i s t o l o g i c a l s t u d y c a n show  of a g i n g  these.  young  are recorded f o r  in interacinar tissue  of a c i n a r t i s s u e .  this  never  t h a n t h a t of the average  In f a c t the g r e a t e s t weights  5 t o 9 month o l d g l a n d s  1 and 2  of  steady during  d e p e n d i n g on t h e i n d i v i d u a l a n i m a l , i t  declines to a value  Perhaps  d i d n o t become  21).  Despite aging,  the o l d e r female glands  Unlike  are  prominent.  elements  compen-  conclusion,  a  which occur animals.  Only  -27-  Physiology Data on p r e p u t i a l gland weights were c o l l e c t e d f o r each animal i n the experimental groups i n order to study the p o s s i b l e e f f e c t of hormonal measurement.  s t i m u l a t i o n on t h i s  physical  As d e s c r i b e d p r e v i o u s l y i n the s e c t i o n on  M a t e r i a l s and Methods, the two glands from each mouse of the adult and weanling groups ( I , I i ,  and II) were weighed  s i n g l y so that a comparison of weights, before and a f t e r treatment, f o r each animal could be made.  However, such a  comparison could not be made i n the newborn group  (ill)  since the p a i r e d glands from each mouse of t h i s l a t t e r  group  had to be weighed together a f t e r treatment and then averaged. The average weights of s i n g l e male glands and of s i n g l e female glands i n each subgroup of groups I, I i ,  II and I I I  were c a l c u l a t e d , so t h a t i t was p o s s i b l e to make a comparison of average weights between the subgroups w i t h i n a group. In a d d i t i o n to the measurement of p r e p u t i a l gland weights, the body weight of each animal was measured before ( I , I i , and II animals) and a f t e r the experiment ( a l l groups). This made i t p o s s i b l e to determine, f o r each mouse, the r e l a t i v e p r e p u t i a l gland weight which denotes mg. per 10 gm. was  of body weight.  of gland  R e l a t i v e p r e p u t i a l gland weight  considered to be a more meaningful measurement than  absolute p r e p u t i a l gland weight because of the normal range of v a r i a t i o n which i s found among mice of the same age and which seems to be p a r t i a l l y r e l a t e d to a s i m i l a r v a r i a t i o n  -28-  i n body w e i g h t among s u c h a n i m a l s g i v e s the average (all  groups)  analysed using  analysis  multiple  r e l a t i v e weights  comparisons  i n Table IV.  of the g l a n d s i s  r e l a t i v e weight  sample  of t h e  the average  distinctly positive propionate,  female v e h i c l e weight  differences  program  loss,  weight  average subgroups  treatment)  a l l of w h i c h  But  there c_, and  produced  w i t h t h e e x c e p t i o n of  c o n t r o l g r o u p c w h i c h showed a  g a i n anyway.  IV,  treatment a,  d i f f e r e n c e b e t w e e n t r e a t m e n t s b,  r e l a t i v e weight  the  a f t e r the  changes of g l a n d s of the c o n t r o l  d upon the p r e p u t i a l g l a n d w e i g h t s ,  for  from  s i g n i f i c a n t l y d i f f e r e n t from the  significant  an a v e r a g e  test  relative  f o r b o t h male and f e m a l e m i c e w i t h i n t h e g r o u p . no  a  With reference to Table  d and c_, and t h e s u b g r o u p b_ ( a f t e r e s t r a d i o l  is  The  s i z e , to t e s t  The r e s u l t s  a d m i n i s t r a t i o n of t e s t o s t e r o n e therefore  collected.  to determine the s i g n i f i c a n t  i n t h e a d u l t c a s t r a t e g r o u p I,  and i s  p r e p u t i a l glands  t e s t p r o v i d e d the F v a l u e s  between t r e a t m e n t subgroups.  change  of s i n g l e  III  a computer program f o r  w i t h unequal  This  w h i c h i t was p o s s i b l e  summarized  Table  of v a r i a n c e , which used S c h e f f e ' s  t r e a t m e n t means.  are  II).  compiled from the d a t a , thus  d a t a were a l s o one-way  (Table  the  negligible  In the i n t a c t a d u l t group I i  t h e r e were  no s i g n i f i c a n t d i f f e r e n c e s b e t w e e n t r e a t m e n t g r o u p s f o r male mice but comparing g r o u p I,  i t is  this  group t o males  of t h e a d u l t  i n t e r e s t i n g t o note t h a t the  castrate  administration  of e s t r a d i o l b e n z o a t e , t r e a t m e n t b_, e f f e c t e d a s i m i l a r  -29-  TABLE I I I AVERAGE RELATIVE WEIGHTS OP SINGLE PREPUTIAL GLANDS  Group  Treatment Subgroup  a)Testosterone propionate  ii; intact weanling mice  Relative I n i t i a l P r e p u t i a l Weight (mg/lOgm)  Relative Final P r e p u t i a l Weight (mg/lOgm)  M 5 F 5  14.6 1.1  18.0 8.0  M 5 F 3  12.7 1.2  7.5 1.0  M 5 F 5  13.4 0.9  4.8 1.0  d)Control  M 2 F 4  14.5 1.2  6.2 1.0  a)Te s t o s t e r o n e propionate  M 3 F 3  12.8 0.5  13.1 11.8  b)Estradiol benzoate  M 3 F 2  11.9 0.6  7.0 0.3  •c) Sesame O i l U.S.P.  M 1 F 1  13.3 1.9  16.2 0.7  d)Control  M 7 F 6  15.6 1.1  14.2 1.6  a)Te s t o s t e r o n e propionate  M 3 F 5  2.0 1.0  16.9 9.2  b)Estradiol benzoate  M 4 F 5  1.9 0.9  1.4 0.9  c)Sesame O i l U.S.P.  M 5 F 6  1.8 0.8  14.2 1.1  d)Control  M 5 F 6  2.2 0.9  10.7 1.4  b)Estradiol i; benzoate castrated c)Sesame O i l adult mice U.S.P.  ii; intact adult mice  Sex and No.  -30-  TABLE I I I c o n t .  Group  Treatment Subgroup a)Te s t o s t e r o n e propionate  b)Estradiol III; benzoate intact newborn c)Sesame O i l mice U.S.P. d)Control  Sex and No.  Relative I n i t i a l P r e p u t i a l Weight (mg/lOgm)  Relative Pinal P r e p u t i a l Weight (mg/lOgm)  M 4 F 6  5.5 4.0  M 3 F 4  1.2 1.6  M 4 F 5  1.8 1.3  M 17 F 22  — — _  2.0 0.9  The f i g u r e s f o r t h e c o n t r o l s i n g r o u p I I I a r e c o m p i l e d f r o m the a v e r a g e i n i t i a l r e l a t i v e p r e p u t i a l w e i g h t s o f a l l mice i n g r o u p I I .  NOTES TO denotes male; F denotes  female  See T a b l e I f o r f u r t h e r  details  TABLE IV  The f i r s t f i g u r e i n e a c h b l o c k o f g r o u p I , I i , and I I r e s u l t s i s t h e a v e r a g e r e l a t i v e w e i g h t g a i n o r l o s s (mg/lOgm) p e r p a i r o f g l a n d s : t h i s f i g u r e was d e r i v e d i n t h e f o l l o w i n g manner u s i n g t h e p a i r e d g l a n d s f r o m each a n i m a l . The r e l a t i v e w e i g h t o f t h e g l a n d removed at. t h e o n s e t o f t h e e x p e r i m e n t was s u b t r a c t e d f r o m t h e r e l a t i v e w e i g h t o f t h e g l a n d removed a t t h e end o f t h e e x p e r i m e n t ; t h e sum o f t h e s e d i f f e r e n c e s i n e a c h t r e a t m e n t s u b g r o u p was d i v i d e d b y t h e t o t a l number of a n i m a l s i n t h e s u b g r o u p t o g i v e t h e a v e r a g e r e l a t i v e w e i g h t g a i n o r l o s s as g i v e n i n t h e t a b l e . The s e c o n d f i g u r e error. The is,  i n each b l o c k of r e s u l t s  ( a l l groups) r e p r e s e n t s the s t a n d a r d  t h i r d f i g u r e i n e a c h b l o c k o f r e s u l t s ( a l l g r o u p s ) i s t h e sample s i z e ; t h e number o f a n i m a l s i n t h e s u b g r o u p .  that  The f i r s t f i g u r e i n e a c h b l o c k o f g r o u p I I I r e s u l t s i s t h e a v e r a g e r e l a t i v e f i n a l p r e p u t i a l g l a n d w e i g h t (mg/lOgm). The a v e r a g e r e l a t i v e w e i g h t g a i n o r l o s s p e r p a i r o f g l a n d s was n o t c a l c u l a t e d f o r t h i s g r o u p s i n c e t h e l e f t p r e p u t i a l g l a n d was n o t removed a t t h e b e g i n n i n g o f t h e e x p e r i m e n t as i n g r o u p s I , I i , and I I . T h e r e f o r e w e i g h t s b e f o r e and a f t e r e a c h t r e a t m e n t c o u l d n o t be compared. I n s t e a d t h e p a i r e d g l a n d s f r o m e a c h mouse o f g r o u p I I I were w e i g h e d t o g e t h e r and t h e n a v e r a g e d . The f i g u r e s f o r t h e c o n t r o l s i n g r o u p I I I a r e c o m p i l e d f r o m the a v e r a g e r e l a t i v e p r e p u t i a l w e i g h t s of a l l mice i n group I I . indicates indicates indicates indicates indicates  significant significant significant significant significant  difference difference difference difference difference  from from from from from  treatment treatment treatment treatment treatment  a a b b c  at at at at at  the the the the the  5% 1% 5% 1% 5%  level, level, level, level, level.  initial  TABLE  IV  COMPUTER PROGRAM ANALYSIS OF THE PHYSIOLOGICAL STUDY  Treatment  3 ,  Group I : c a s t r a t e d Young a d u l t m i c e F  M a)Te s t o sterone propionate  3.4  6.9  b  ±1.5  C  (5)  d  M b  ±0.8° (5)  Group I i : I n t a c t Young a d u l t mice  d  F  Group I l r i n t a g t Weanling Mice M  F  Group III: I n t a c t Newborn a mice F  M  0.3  11.3  14.9.  8.2  5.5  ±3.8  ±2.5  ±0.5  ±0.5  ±0.6  0.3  (3)  (5)  (4)  (6)  1.2**  1.6**  (3)  (3)  4.0  e  e  b Estradiol benzoate  -5.2*  -0.2**  -5.0  -0.3**  -0.4**  -0.04**  ±0.5  ±0.2  ±3.1  ±0.1  ±0.2  ±0.1  ±0.3  ±0.3  (5)  (3)  (3)  (2)  (4)  (5)  (3)  (4)  c)Sesame Oil,U.S.P. vehicle control  -8.6**  0.2**  1.8**  1.3**  d)Control  12.4''  0.03**  ±2.2  ±0.1  ±1.0  ±0.2  ±0.2  ±0.1  (5)  (5)  (5)  (6)  (4)  (5)  -8.3*  -0.2**  -1.5  ±2.6  ±0.1  ±1.1  ±0.3  ±0.7  ±0.2  ±0.1  0.9 ' ±0.05  (2)  (4)  (7)  (6)  (5)  (6)  (17)  (22)  0.5**  8.5**, z  0.5**  1.9  f  '  * *  f  t!  -33-  average r e l a t i v e weight l o s s i n both groups  (-5.2 mg/lO gm  f o r group IbM; -5.0 mg/lO gm f o r group IibM).  Furthermore  there i s a s t r i k i n g d i f f e r e n c e between the average  relative  weight changes of the c o n t r o l groups; the c a s t r a t e d show a much greater weight l o s s . i n t a c t a d u l t group I i ,  controls  F o r the females of the  the a d m i n i s t r a t i o n of t e s t o s t e r o n e  propionate e f f e c t e d a d i s t i n c t weight g a i n i n the glands of subgroup a (11.3 mg/lO gm) which i s s i g n i f i c a n t l y  different  at the 1% l e v e l from the weight changes of glands of treatment subgroups b and d.  In the weanling group I I the average  r e l a t i v e weight g a i n of male p r e p u t i a l glands (14.9 mg/lO gm) a f t e r the a d m i n i s t r a t i o n of t e s t o s t e r o n e propionate i s s i g n i f i c a n t l y d i f f e r e n t from the weight changes of male c o n t r o l glands (8.5 mg/lO gm) and of e s t r a d i o l  treated  glands (-0.4 mg/lO gm), but i s not s i g n i f i c a n t l y  different  from the average r e l a t i v e weight g a i n (12.4 mg/lO gm) which a l s o occurs i n glands of the v e h i c l e c o n t r o l subgroup c. In f a c t there i s a s i g n i f i c a n t d i f f e r e n c e at the 5% l e v e l between the weight changes of the two c o n t r o l subgroups  which  i s i n e x p l i c a b l e to me although i t i s probably not a t t r i butable to the sesame o i l i n j e c t i o n s  (since no such  d i f f e r e n c e occurs between c o n t r o l s of the other groups, male or f e m a l e ) .  What i s more important i n the male weanling  group i s the s i g n i f i c a n t d i f f e r e n c e which occurs between the average r e l a t i v e weight changes  i n glands of both  c o n t r o l subgroups and the e s t r a d i o l t r e a t e d glands.  The  -34-  latter  show an a v e r a g e  i n the w e a n l i n g group  r e l a t i v e weight  at  a (8.2 mg/lO gm)  which  relative  a, a f t e r  b, c, and d.  different changes  Finally,  I I I , f o r b o t h m a l e s and f e m a l e s , t h e  final  preputial  weight  of glands i n  preputial  weight  and  subgroup  no  significant  b  upon female  from the average  relative  of glands i n the c o n t r o l  (after  estradiol  final  subgroups  treatment).  gland weights.  final  subgroups variations  preputial  The  significant  c a n be r e a d i l y i n subgroup  weights of female  d anima,ls.  average  glands i n these  I f these f i g u r e s  weight are  seem h i g h compared t o t h e b o d y  o f f e m a l e mice i n t h e o t h e r s u b g r o u p s preputial  d  difference,  a c c o u n t e d f o r by t h e b o d y  examined, the body w e i g h t s  relative  and  g l a n d w e i g h t s , and t r e a t m e n t s b an _c  t h e 5% l e v e l b e t w e e n t r e a t m e n t s b and d upon t h e  relative  c, d  But t h e r e i s  d i f f e r e n c e b e t w e e n t r e a t m e n t s b, c,  u p o n male p r e p u t i a l  weights  subgroup  the a d m i n i s t r a t i o n of t e s t o s t e r o n e p r o p i o n a t e , i s  significantly different  at  g a i n i n glands of  r e l a t i v e weight  i n glands of t r e a t m e n t subgroups  average  females  is significantly  t h e Yfo l e v e l f r o m t h e a v e r a g e  i n t h e newborn g r o u p  For  the a d m i n i s t r a t i o n of t e s t o s t e r o n e  propionate e f f e c t e d a d i s t i n c t weight subgroup  loss.  so t h a t  the  w e i g h t s of d a n i m a l s are c o r r e s p o n d i n g l y  affected. U s u a l l y , but not always, the weight g l a n d s of each t r e a t m e n t subgroup  changes i n t h e  are i n d i c a t i v e  of  their  -35-  present p h y s i o l o g i c a l s t a t e and microscopic s t r u c t u r e . Therefore during the c o n s i d e r a t i o n of the microscopic appearance of; the t r e a t e d p r e p u t i a l glands which f o l l o w s , I s h a l l c o n s t a n t l y r e f e r to Table IV and the  relationship  between p h y s i c a l measurements and h i s t o l o g y or p h y s i o l o g y .  Group I;  c a s t r a t e d a d u l t mice  Male s The  l e f t p r e p u t i a l glands which were removed from  each 7 week male mouse at the onset of the experiment e x h i b i t the f u l l y developed, p a t t e r n ( F i g . 23) which was previous s e c t i o n .  compound a l v e o l a r  structural  described i n d e t a i l i n a  Of course, there are s l i g h t  individual  v a r i a t i o n s i n p r e p u t i a l gland s i z e , i n the number of accessory ducts which course through the parenchyma of the gland and  i n the amount of i n t e r a c i n a r and  connective t i s s u e . animals  subepithelial  In the 10 week c a s t r a t e d c o n t r o l  (subgroups c_, d) the p r e p u t i a l glands  degree of atrophy  ( F i g . 24).  show some  G e n e r a l l y there i s a  i n the o v e r a l l s i z e of the glands.  In. the l i t t l e  decrease remaining  parenchyma i t i s s t i l l p o s s i b l e to d i s t i n g u i s h the v a r i o u s types of a c i n i which make up the c y c l e of sebum formation but there are g e n e r a l l y fewer c e l l s than usual i n each acinus  ( F i g . 28).  The  duct system i s a l s o l e s s e x t e n s i v e .  Furthermore there i s an i n c r e a s e i n the stromal framework of the gland, manifest by an abundant f i b r o u s i n t e r a c i n a r  -36-  and  s u b e p i t h e l i a l stroma and numerous f i b r o u s t r a b e c u l a e .  A l l these  s t r u c t u r a l changes give the glands  f e m a l e - l i k e microscopic p a t t e r n and  a somewhat  i n d i c a t e an  overall  depression of s e c r e t o r y a c t i v i t y which a l s o e x p l a i n s the v e r y d e f i n i t e post c a s t r a t i o n decrease gland  i n r e l a t i v e weight of the  (Tables I I I and.IV). The  d a i l y a d m i n i s t r a t i o n of t e s t o s t e r o n e  propionate  f o r three weeks to c a s t r a t e d male mice maintains s t r u c t u r a l p a t t e r n of the p r e p u t i a l gland and  the a d u l t  thereby  counteracts the post c a s t r a t i o n p r e p u t i a l g l a n d u l a r  atrophy  which was  signi-r-  observed  i n the c o n t r o l glands; hence the  f i c a n t d i f f e r e n c e between the c o n t r o l subgroups -c_ and d and a ( a f t e r t e s t o s t e r o n e treatment) on the b a s i s of r e l a t i v e weight (Table I V ) .  But  s t r u c t u r a l l y the t r e a t e d glands  not s t i m u l a t e d by the hormone to the same degree.  are  In three  of the t r e a t e d glands the s t i m u l a t o r y e f f e c t i s manifest m i c r o s c o p i c a l l y by h y p e r p l a s i a of the a c i n a r c e l l s and i n c r e a s e d d u c t u l a r t r a n s f o r m a t i o n of tb,e parenchyma.  by The  l a t t e r i s so e x t e n s i v e , that i n s p i t e of the h y p e r p l a s i a , the a c i n i are mainly  confined to the p e r i p h e r y of the  between the e p i t h e l i u m of the ducts and the t i s s u e capsule treatment  ( F i g . 26).  group are c y s t i c  The  remaining  two  connective glands  ( F i g . 27), resembling  11 and 12 month o l d male glands.  The  gland  c e n t r a l and  the  in this aging  lateral  ducts are f i l l e d w i t h sebum, lymphocytes, polymorphonuclear n e u t r o p h i l e s , and desquamated duct e p i t h e l i a l c e l l s .  Acini,  -37-  stroma  and even duct e p i t h e l i u m degenerate  by numerous lymphocytes  ( F i g . 29).  as they are invaded  At the p e r i p h e r y of the  gland only a few compact masses of a c i n i remain.  Here the  c y s t i c atrophy i s probably "the r e s u l t of o v e r s t i m u l a t i o n by the repeated a c t i o n of the hormone during the three week i n j e c t i o n p e r i o d which leads to what Beaver (i960) c a l l s "exhaustion atrophy"; h y p e r p l a s i a of the a c i n a r c e l l s  cannot  keep up w i t h the c e l l u l a r d e s t r u c t i o n which of n e c e s s i t y accompanies h o l o c r i n e s e c r e t i o n . Finally,  i n c a s t r a t e d male mice a f t e r the d a i l y  a d m i n i s t r a t i o n of e s t r a d i o l benzoate  f o r three weeks, the  p r e p u t i a l glands show a s i m i l a r degree c o n t r o l glands  (of subgroups £ and d ) .  of atrophy as do the The glands are  reduced i n s i z e both g r o s s l y , on the b a s i s of r e l a t i v e weight  (Table I V ) , and m i c r o s c o p i c a l l y ; there i s l i t t l e  remaining parenchyma which g e n e r a l l y c o n s i s t s of small a c i n i , an abundant f i b r o u s i n t e r a c i n a r and and a l e s s extensive duct system  subepithelial  ( F i g . 25.).  stroma  Since these  changes, which give the glands the appearance of aging female  glands, and i n d i c a t e a d e p r e s s i o n of s e c r e t o r y a c t i v i t y ,  a l s o occur i n the glands of male c a s t r a t e d c o n t r o l s , they may  be a t t r i b u t a b l e mainly to the absence of the t e s t e s  r a t h e r than to the presence  of the e s t r o g e n i c hormone.  However, there are f u r t h e r changes i n the e s t r a d i o l glands of c a s t r a t e d males which are perhaps  treated  directly  e f f e c t e d by the hormone; the thickened s t r a t i f i e d e p i t h e l i u m of the l a t e r a l and c e n t r a l ducts; the presence of many  -38-  desquamated e p i t h e l i a l c e l l s i n the ducts ( F i g . 30).  Females The l e f t p r e p u t i a l glands which were removed from 7 week female mouse a t the onset of the experiment  each  exhibit,  w i t h s l i g h t i n d i v i d u a l v a r i a t i o n s , of course, the t y p i c a l s t r u c t u r a l p a t t e r n of the young a d u l t female gland ( F i g . 31) which has been p r e v i o u s l y d e s c r i b e d . 10 week female glands i n subgroups  A f t e r c a s t r a t i o n , the  c_ and d are not a t r o p h i c  ( F i g . 32), u n l i k e the glands of male c a s t r a t e d c o n t r o l s .  In  the female, gross (on the b a s i s of r e l a t i v e weight) and microscopic d i f f e r e n c e s are n e g l i g i b l e .  A few obvious  aging e f f e c t s are observed but even these v a r y from animal to animal; a decrease i n the amount of parenchyma, smaller a c i n i , a more abundant stroma, the i n v a s i o n of some a c i n i by connective t i s s u e elements, f o r example. A f t e r the d a i l y a d m i n i s t r a t i o n of t e s t o s t e r o n e propionate f o r three weeks to c a s t r a t e d female mice, the p r e p u t i a l glands show a s i g n i f i c a n t i n c r e a s e i n average r e l a t i v e weight  (Table I V ) .  M i c r o s c o p i c a l l y , the s t i m u l a t o r y  e f f e c t of the hormone i s manifest by h y p e r p l a s i a of the paren-n chymal c e l l s and by hypertrophy of the a c i n i . a c i n i appear  l a r g e r than i n the 7 week female glands or i n  the 10 week c o n t r o l glands. conglomerates  Therefore the  of a c i n i  Furthermore,  the frequenpy of  (types 4 and 5) which are undergoing  degeneration, has i n c r e a s e d ; the duct system has become  -39-  more e x t e n s i v e b y d u c t u l a r t r a n s f o r m a t i o n , and o n l y delicate (Figs.  stroma of c o n n e c t i v e t i s s u e  35 and 3 6 ) .  Because  w h i c h are the r e s u l t  of these  of t e s t o s t e r o n e  female glands b e g i n to resemble  surrounds the structural  a acini  changes,  stimulation,  the  t h e more a c t i v e l y  secreting  g l a n d s o f i n t a c t a d u l t male m i c e . After  e s t r a d i o l treatment the glands of  c a s t r a t e d mice g e n e r a l l y resemble c o n t r o l female's effects  But u n l i k e  t h e g l a n d s o f the! u n t r e a t e d  ( i n s u b g r o u p s £ and d) ; t h e o b v i o u s  are observed-the  parenchyma,  female  the i n c r e a s e  r e d u c t i o n i n t h e amount  of  i n the s t r o m a l network  (Fig.  t h e c o n t r o l g l a n d s and n o t u n l i k e  the  t h e r e are changes i n t h e s t r u c t u r e female glands which are perhaps  epithelial the d u c t s  Group I i ;  of t h e e s t r a d i o l  treated the  t h i c k e n e d and t h e  c e l l s seem t o be c o n t i n u o u s l y (Fig.  animals,  d i r e c t l y e f f e c t e d by  the d u c t u l a r e p i t h e l i u m i s  sloughed  off  serve  i n t a c t a d u l t mice  as a s o u r c e  estrogenic  of comparison,  of group  group  and  similarly treated  I.  c o n t r a s t t o the post  i n the c o n t r o l males  of  a f t e r androgenic  t r e a t m e n t s , to the glands of the  c a s t r a t e d animals In  into  34).  The p r e p u t i a l g l a n d s o f i n t a c t , a d u l t a n i m a l s Ii  33).  condition  i n t h e e s t r a d i o l t r e a t e d g l a n d s o f c a s t r a t e d male  hormone;  aging  o f g r o u p I,  castration glandular  atrophy  the p r e p u t i a l glands  of  -40-  group I i male c o n t r o l s continue to e x h i b i t the mature microscopic p a t t e r n ( F i g . 37). observed  - the lymphocytic  A few aging e f f e c t s may  i n f i l t r a t i o n of the  be  connective  t i s s u e stroma, the i n v a s i o n and subsequent degeneration of some a c i n i and the e p i t h e l i u m of some ducts by and other connective t i s s u e elements. observed  lymphocytes  S i m i l a r r e s u l t s are  i n the glands of group I i c o n t r o l females  a l s o l i k e the glands of group I c o n t r o l females,  which,  continue  to e x h i b i t the mature female microscopic p a t t e r n ( F i g . 41). Again aging e f f e c t s may  be observed - a decrease  i n the  amount of parenchyma, an increase i n the lumen of the  central  duct, smaller a c i n i , a more abundant stroma, the i n v a s i o n of some a c i n i by connective t i s s u e elements ( F i g . 42). A f t e r the a d m i n i s t r a t i o n of t e s t o s t e r o n e propionate to i n t a c t male animals the p r e p u t i a l glands show n e g l i g i b l e gross d i f f e r e n c e s on the b a s i s of r e l a t i v e weight and when compared to c o n t r o l glands, almost structural differences.  The  (Table  IV)  negligible  l a t t e r , however, are  difficult  to assess o b j e c t i v e l y since n o t i c e a b l e v a r i a t i o n s i n the amount of parenchyma and the extent of the duct system, f o r example, occur among the i n d i v i d u a l glands of t h i s subgroup.  treatment  There does seem to be an i n d i c a t i o n of i n c r e a s e d  s e c r e t o r y a c t i v i t y i n one  of the glands, as manifest by  h y p e r p l a s i a of the a c i n a r c e l l s and i n c r e a s e d d u c t u l a r t r a n s f o r m a t i o n of the parenchyma ( F i g . 38.). t h i s , the s t i m u l a t i o n of female  In c o n t r a s t to  glands i n i n t a c t  animals  -41-  receiving testosterone  is  s i g n i f i c a n t l y greater both  on t h e b a s i s o f r e l a t i v e w e i g h t  (Table  The s t i m u l a t o r y e f f e c t o f t h e hormone is manifest microscopically,  IV)  grossly,  and m i c r o s c o p i c a l l y .  i n the female  gland  i n much t h e same way as  i n the  g l a n d s of group l a c a s t r a t e d f e m a l e s - by h y p e r p l a s i a the parenchymal  c e l l s , hypertrophy  of the a c i n i ,  and by  i n c r e a s e d d u c t u l a r t r a n s f o r m a t i o n , a l l of which give gland a m a l e - l i k e s t r u c t u r a l p a t t e r n (Fig. There group I ment.  is  a resemblance  and t h o s e Although  of group I i  group I i  (on t h e b a s i s  (Fig.  which g e n e r a l l y  consists  fibrous  are  a s t r i k i n g decrease  39)  structural  duct system  - Table IV)  there i s  little  remaining an  parenchyma  abundant  less the  s i m i l a r i t y between the glands of these  two  i n the c a s t r a t e a n i m a l s , of the g l a n d ,  i n the i n t a c t a n i m a l s ,  given i n t h i s  experiment,  gland, possibly  Whereas  does n o t b r i n g  about the  i n group I i  endocrine  the m i c r o s c o p i c  p r e p u t i a l gland g e n e r a l l y resembles  atrophy doses  an e f f e c t u p o n t h e  the a d m i n i s t r a t i o n of e s t r a d i o l benzoate  female animals  estradiol,  e s t r a d i o l , i n the  seems t o have  by a l t e r i n g e x i s t i n g  grossly  and  The r e a s o n s f o r  n o t t h e same.  treatglands  in size  of s m a l l - s i z e d a c i n i ,  (Fig. 40).  subgroups are o b v i o u s l y  of  i n t a c t , t h e male  i n t e r a c i n a r and s u b e p i t h e l i a l s t r o m a , a  extensive  After  43).  a f t e r e s t r a d i o l benzoate  of r e l a t i v e w e i g h t  microscopically  the  b e t w e e n t h e male g l a n d s  animals  are a t r o p h i c t o o , showing  of  relationships. to  intact  structure  t h a t of the  of  control  the  -42-  glands,  w i t h the e x c e p t i o n of changes perhaps  i n d i r e c t l y e f f e c t e d by the e s t r o g e n i c  d i r e c t l y or  hormone;  the  o f t h e squamous e p i t h e l i u m o f some d u c t s  and t h e  of e p i t h e l i a l  44).  Group I I :  c e l l s i n t o the ducts  veanling  (Fig.  metaplasia  desquamation  mice  Males The l e f t p r e p u t i a l g l a n d s w h i c h were removed e a c h 3 week male mouse a t t h e o n s e t compact s t r u c t u r e s  from  of the e x p e r i m e n t ,  m i c r o s c o p i c a l l y w h i c h have n o t  yet  a t t a i n e d the a d u l t h i s t o l o g i c a l p a t t e r n a l t h o u g h  the  a r e d e n s e l y p a c k e d t o g e t h e r and a r e r e c o g n i z a b l e  as  structural  types  of the a d u l t gland  t h e most numerous,  have few c e l l s ; t y p e s  more c e l l s i n t h e m ) . cells  of the a c i n i .  (Fig. 45).  increase  A c e n t r a l duct i s  i n average  s t r u c t u r a l pattern; there :  enlarged a c i n i of types  (Fig.  3, 4,  usually  acini the  are  5 have i n the  r e l a t i v e weight  2, 3 , 4 ,  III  mature  composed  5 w h i c h now have  i n them and t h e d u c t s y s t e m i s  show  (Tables  the glands e x h i b i t the i s much p a r e n c h y m a  basal  present  the g l a n d s of c o n t r o l animals  and I V ) . M i c r o s c o p i c a l l y ,  more c e l l s  2, w h i c h  There a r e f r e q u e n t m i t o s e s  A t 6 weeks  a striking  (type  are  of  many  extensive  46). The e f f e c t o f t h e d a i l y a d m i n i s t r a t i o n o f  propionate difficult  for  19 d a y s t o w e a n l i n g m i c e  to assess.  Grossly,  there i s  testosterone  ( s u b g r o u p a) an i n c r e a s e  is in  -43-  r e l a t i v e w e i g h t and s i z e b u t g e n e r a l l y t h e i n c r e a s e i s no greater than t h a t observed i n c o n t r o l glands (which i s the  r e s u l t o f a g i n g ) f o r , as n o t e d p r e v i o u s l y , t h e  average  r e l a t i v e weight gain of testosterone t r e a t e d glands i s s i g n i f i c a n t l y d i f f e r e n t from the average of  control  subgroup  d glands but i s not  r e l a t i v e weight  gain  significantly  d i f f e r e n t from the average r e l a t i v e w e i g h t g a i n of glands i n the v e h i c l e in  c o n t r o l subgroup £  (Table I V ) .  Microscopically,  some o f t h e g l a n d s t h e r e seems t o be a n i n d i c a t i o n  of  i n c r e a s e d s e c r e t o r y a c t i v i t y as m a n i f e s t b y h y p e r p l a s i a o f the  acinar cells The  ( F i g . 47).  e f f e c t o f e s t r a d i o l b e n z o a t e , a f t e r 19 d a y s  of  t r e a t m e n t , i s , on t h e o t h e r h a n d , most n o t i c e a b l e , b o t h grossly the  ( T a b l e s I I I and I V ) and m i c r o s c o p i c a l l y b e c a u s e  s m a l l amount o f p a r e n c h y m a i n t h e g l a n d s o f s u b g r o u p  a n i m a l s compared t o t h a t i n c o n t r o l g L a n d s . the  parenchyma c o n s i s t s of s m a l l a c i n i  w h i c h have few c e l l s the  abundant  i n them.  s t r o m a w h i c h now  not e x t e n s i v e .  As t h e r e s u l t  b  Furthermore  (of a l l types)  Some a c i n i a r e i n v a d e d b y s u r r o u n d s t h e a c i n i and  b e n e a t h t h e e p i t h e l i u m of t h e d u c t s .  The  lies  duct system i s  of p r o l o n g e d e s t r o g e n i c  t r e a t m e n t the g l a n d i s a t r o p h i c ; the normal growth is  of  pattern  i n h i b i t e d b y t h e hormone t r e a t m e n t ( F i g . 4 8 ) .  Female s S t r u c t u r a l l y the l e f t p r e p u t i a l g l a n d which  was  removed f r o m e a c h 3 week f e m a l e mouse a t t h e b e g i n n i n g o f  -44-  the experiment, resembles (Fig. 49).  t h a t o f t h e male  But a t 6 weeks  a f t e r sexual differences  a p p e a r e d , t h e g l a n d s of c o n t r o l f e m a l e s increase mature  in size.  weanling  Microscopically,  show o n l y a  s t r u c t u r a l p a t t e r n of the f e m a l e ; g e n e r a l l y the  which are undergoing  dissolution  t h e c e n t r a l d u c t and a c c e s s o r y  (type  parenchyma finally, fibrous  is  g l a n d s undergo  w e i g h t and s i z e  (Fig.  by h y p e r p l a s i a  a striking  the gland;  again manifest  the  in relative (Table  IV).  microscopically  c e l l s , hypertrophy  of  and b y i n c r e a s e d d u c t u l a r t r a n s f o r m a t i o n , a l l  of  which give  of the parenchymal  and t h e  propionate  (treatment a), increase  numerous;  50).  compared t o t h e c o n t r o l g l a n d s  The s t i m u l a t o r y e f f e c t i s  (Fig.  so t h a t  the a d m i n i s t r a t i o n of t e s t o s t e r o n e  19 d a y s t o w e a n l i n g f e m a l e m i c e  preputial  acini,  connective tissue  t r a b e c u l a e a r e more a b u n d a n t  acini  developed  c o n f i n e d t o the p e r i p h e r y of the  the i n t e r a c i n a r f i b r o u s  After  of  5) a r e more  ducts are w e l l  and may o c c u p y t h e c e n t r e o f t h e g l a n d  acini,  slight  the glands e x h i b i t the  have many more c e l l s i n them and c o n g l o m e r a t e s  for  have  the gland a m a l e - l i k e s t r u c t u r a l  the  pattern  51). E s t r a d i o l b e n z o a t e , a f t e r 19 days of t r e a t m e n t ,  has  a s i m i l a r e f f e c t m i c r p s c o p i c a l l y upon t h e f e m a l e g l a n d s group II group.  animals  as  i t does u p o n t h e male g l a n d s i n  The p a r e n c h y m a  w h i c h have  consists  few c e l l s i n t h e m .  of s m a l l a c i n i A v e r y abundant  of  this  (of a l l stroma  types)  -45-  surrounds the a c i n i ; connective tissue not v e i l  some a c i n i a r e i n v a d e d b y t h e  elements  of the stroma.  The d u c t s  developed; the d u c t u l a r e p i t h e l i u m i s  and e p i t h e l i a l  c e l l s seem t o be c o n t i n u o u s l y  i n t o the ducts  (Fig.  Group I I I :  often  newborn mice  c o n t r o l g l a n d s of group I I I newborn a n i m a l s  animals  initially  serve  a t 3 weeks.  to serve  as v e h i c l e c o n t r o l s .  a n i m a l s , b o t h male and f e m a l e , have p a t t e r n of t h e w e a n l i n g mouse;  A few  control  a compact mass o f  structural acini  as t h e t y p e s  of  which surrounds a small c e n t r a l duct.  The d a i l y a d m i n i s t r a t i o n o f t e s t o s t e r o n e t o newborn mice f o r 3 weeks has the average  the  Microscopically,  t h e immature  small i n s i z e but recognizable  the a d u l t gland)  as  r e c e i v e d a d a i l y i n j e c t i o n of sesame o i l  t h e n , the p r e p u t i a l g l a n d s of a l l the group I I I  (generally  off  52),  f r o m t h e male and f e m a l e m i c e o f g r o u p I I  3 weeks  thickened  sloughed  The p r e p u t i a l g l a n d s w h i c h were removed  for  are  a significant  propionate  e f f e c t upon  r e l a t i v e w e i g h t o f t h e p r e p u t i a l g l a n d s of  male and f e m a l e m i c e  (Table  b o t h male and f e m a l e g l a n d s , m a n i f e s t by h y p e r p l a s i a  IV).  Microscopically,  cells  o f many c o n g l o m e r a t e s  is  and  i n c r e a s e d d u c t u l a r t r a n s f o r m a t i o n of the parenchyma i n d i c a t e d by the presence  in  the s t i m u l a t o r y e f f e c t  of the parenchymal  both  as  of type 4  _46-  and 5 a c i n i and by the appearance of l a t e r a l ducts (Figs. 53 and 54). In the m a j o r i t y of the treated glands the most numerous a c i n i are s t i l l the small type 2 a c i n i which are t y p i c a l l y present i n the c o n t r o l glands.  However,  two of the treated male glands show a greater response to the hormone, f o r there has not only been hyperplasia of the acinar c e l l s but hypertrophy of many of the a c i n i as w e l l ( F i g . 58) so that the glands begin to e x h i b i t the adult s t r u c t u r a l pattern. A f t e r the d a i l y a d m i n i s t r a t i o n of e s t r a d i o l benzoate to newborn mice f o r 3 weeks, the male and female p r e p u t i a l glands continue to resemble, g e n e r a l l y , the normal glands of weanling mice, with the exception of a few s t r u c t u r a l d e t a i l s , which are not widespread; the more abundant stroma, the thickened epithelium l i n i n g the c e n t r a l duct, the invasion of some a c i n i by connective t i s s u e  elements  (Figs 55 and 56). Summary of the e f f e c t s of the p h y s i o l o g i c a l treatments The mouse p r e p u t i a l gland responds d i r e c t l y to testosterone propionate.  The stimulatory e f f e c t i s  manifest m i c r o s c o p i c a l l y most often by hyperplasia of the parenchymal c e l l s and o c c a s i o n a l l y by hypertrophy of the a c i n i (usually i n the female glands).  Therefore mitoses,  -47-  p a r t i c u l a r l y i n t h e w e a n l i n g and n e w b o r n a n i m a l s I l a and  Ilia  cells.  S t i m u l a t i o n of the female  r e s p e c t i v e l y ) are f r e q u e n t i n the b a s a l a c i n a r  o f t h e male g l a n d s  glands i s greater than that  i n a l l subgroups r e c e i v i n g t e s t o s t e r o n e  w i t h t h e e x c e p t i o n of t h e newborn The  (groups  group.  mouse p r e p u t i a l g l a n d i s n o t s t i m u l a t e d b y  e s t r a d i o l benzoate.  The  a c t u a l response  t h e hormone depends on t h e age prolonged treatment  of t h e g l a n d t o  of t h e a n i m a l .  o f newborn a n i m a l s  After  (group I l l b )  the  m i c r o s c o p i c appearance of the p r e p u t i a l g l a n d s i s l i t t l e a f f e c t e d compared t o t h a t o f n o r m a l age.  The  of t h e same  glands of both subgroups e x h i b i t the  immature p a t t e r n . estradiol  animals  But  overall  the glands of w e a n l i n g animals  ( g r o u p l i b ) do n o t d e v e l o p n o r m a l l y .  The  receiving  effect  i s more n o t i c e a b l e i n male g l a n d s b e c a u s e d u r i n g t h e interval  f r o m 3 weeks t o 6 w e e k s , when n o r m a l l y male  g l a n d s show e v i d e n c e  of a c t i v e  male g l a n d s r e c e i v i n g e s t r a d i o l become a t r o p h i c .  s e c r e t i o n , the growth i s i n h i b i t e d ; the  glands  S i m i l a r l y , the glands of i n t a c t a d u l t  males r e c e i v i n g e s t r a d i o l  ( g r o u p l i b ) show an  d e p r e s s i o n o f s e c r e t o r y a c t i v i t y m a n i f e s t by  overall decreased  d u c t u l a r t r a n s f o r m a t i o n and a c i n a r a t r o p h y a l l o f g i v e the glands the appearance of a g i n g female Additional effects  of  which  glands.  of e s t r a d i o l - t h e t h i c k e n i n g o f t h e  d u c t u l a r e p i t h e l i u m and  the continuous  s l o u g h i n g o f f of  -48-  epithelial in  cells  i n t o the ducts,  f o r e x a m p l e , "which a r e n o t e d  some g l a n d s a f t e r e s t r a d i o l t r e a t m e n t s - w i l l  be  dis-  cussed i n the f o l l o w i n g s e c t i o n of the t h e s i s .  Histochemistry The l i p i d  component o f t h e mouse p r e p u t i a l  gland  i s most c l e a r l y d e m o n s t r a t e d i n f r e s h f r o z e n s e c t i o n s o f t i s s u e b y t h e f a t - s o l u a b l e d y e , Sudan b l a c k B, u s e d as a saturated  s o l u t i o n i n 70% e t h y l a l c o h o l .  b l a c k by the dyej  numerous l i p i d  Coloured  droplets  throughout the cytoplasm o f the a c i n a r  appear  cells.  scattered  The  droplets  seem t o a c c u m u l a t e g r a d u a l l y i n t h e a c i n i ,  gressing  from t i n y d r o p l e t s  gland  t o homogenous  (Fig. 60).  and d e g e n e r a t i n g a c i n i ,  masses o f l i p i d  component o f a d u l t and w e a n l i n g g l a n d s ,  lipid  of e i t h e r sex,  The r e s u l t s o f McManus' Sudan  b l a c k B method f o r compound cerebrosides)  and  i n the ducts of the  ( Q u a l i t a t i v e d i f f e r e n c e s i n the  were n o t n o t i c e a b l e . )  pro-  i n the s m a l l a c i n i , to l a r g e  spheres i n the h y p e r t r o p h i e d finally,  blue-  lipids  were n o t c o n s i s t e n t .  ( p h o s p h a t i d e s and In  counterstained  s e c t i o n s , t h e r e were s o m e t i m e s i n t h e s m a l l a c i n i a few fat  droplets, partially  trophied a c i n i , acinar  cell  stained black  and i n t h e h y p e r -  c l o s e l y packed g l o b u l e s  almost completely.  which f i l l e d  T h e s e , t o o , were  only  each  -49-  h e a v i l y o u t l i n e d by the s t a i n ; they d i d not appear as the dense d r o p l e t s d e s c r i b e d above by the other Sudan technique  (Fig. 6 l ) .  Sudan black B has a strong  f o r most c l a s s e s of l i p i d s  affinity  so that p o s i t i v e r e s u l t s  with  McManus' method do not n e c e s s a r i l y s i g n i f y the presence of phosphatides and are excluded  cerebrosides.  from the l i p i d  Only i f n e u t r a l f a t s  component does the s t a i n become  u s e f u l i n the demonstration of these (Pearse,  1968).  from the l i p i d  compound  lipids  However, n e u t r a l f a t s cannot be  excluded  component of the mouse p r e p u t i a l gland  as  shown by the p o s i t i v e r e s u l t s of o i l red 0 s t a i n i n g by differential  s t a i n i n g technique  of Beaver ( i 9 6 0 ) .  O i l red  0 i s a strong colourant of f a t s but a v e r y poor one phospholipids  the  for  (Chayen, B i t e n s k y , Butcher, P o u l t e r , 1969).  In the mouse p r e p u t i a l gland the c o n c e n t r a t i o n of f a t s i s very great as manifest  by o i l red 0 s t a i n i n g .  Moreover,  the d e p o s i t i o n of o i l red 0 d r o p l e t s f o l l o w s c l o s e l y that revealed by Sudan black B s t a i n i n g i n f r o z e n s e c t i o n s ; the o i l red 0 d r o p l e t s are conspicuous throughout the gland with the l a r g e s t d r o p l e t s c o a l e s c i n g i n the degenerating ( F i g s . 62 and The  a c i n a r c e l l s of the hypertrophied  acini  67).  lipid  component of the gland d i d not  stain  f o l l o w i n g the p e r f o r m i c - a c i d S c h i f f method f o r compound lipids  c o n t a i n i n g unsaturated  bonds (that i s , f o r  -50-  phospholipids  and  c e r e b r o s i d e s ) ; there was  s t a i n i n g of the cytoplasm and  only d i f f u s e  r a t h e r more intense s t a i n i n g  of the n u c l e i (the l a t t e r because of t h e i r DNA The  Schultz method f o r c h o l e s t e r o l and  content).  i t s e s t e r s was  also  negative. The  several histochemical  mouse p r e p u t i a l gland  t e s t s c a r r i e d out on  f o r p r o t e i n s d i d not r e v e a l d e f i n i t e  s t r u c t u r e s of a proteinaceous nature i n the  acinar  cells,  which would c o n t r i b u t e d i s t i n c t l y to the formation sebum, i n a d d i t i o n to the degenerated. proteins  l i p i d component, as the  c o n t a i n i n g t y r o s i n e and  cells  (at pH  9.5)  there was  c o n t a i n i n g amino a c i d s  c y s t e i n e ) by the formation gave r i s e to weak and  only  stroma of  f e r r i c f e r r i c y a n i d e method which should  the presence of sulphur  the reveal  (cystine  of the pigment, P r u s s i a n  blue,  However i n s e c t i o n s of  gland which also i n c l u d e d the epidermis of the  the  prepuce,  the extreme proximal p o r t i o n of the main e x c r e t o r y  of the gland,  and  n o n - s p e c i f i c green s t a i n i n g throughout  the parenchyma of the gland.  and  the  a f t e r the B i e b r i c h s c a r l e t  d i f f u s e s t a i n i n g of the parenchyma and the The  of  In b r i e f , a f t e r the M i l l o n r e a c t i o n f o r  method f o r b a s i c n u c l e o p r o t e i n s  gland.  the  the squamous e p i t h e l i u m  of these  duct  structures  s t a i n e d p o s i t i v e l y , because of t h e i r k e r a t i n content. ( K e r a t i n contains  S-S  and  S-H  groups).  The  i n t e n s i t y of  -51-  r e a c t i o n i n the e p i t h e l i u m of the main e x c r e t o r y duct decreased r a p i d l y from the surface  inwards.  The B i e b r i c h s c a r l e t method of S p i c e r and L i l l i e (at pH 4.95), Heidenhain's M a l l o r y ' s phosphotungstic  i r o n hematoxylin method and a c i d hematoxylin method gave  r i s e only to n o n - s p e c i f i c d i f f u s e s t a i n i n g i n the a c i n a r c e l l s of the mouse p r e p u t i a l gland ( F i g . 66).  In c o n t r a s t ,  these methods demonstrated i n the parenchyma of the r a t p r e p u t i a l gland, the c h a r a c t e r i s t i c cytoplasmic, p r o t e i n a ceous granules, p r e v i o u s l y d e s c r i b e d by Beaver (1959, 1960). There were no  d i f f e r e n c e s between the phosphate f o r m a l i n  f i x e d and the acetate f o r m a l i n f i x e d r a t glands.  With  B i e b r i c h s c a r l e t at pH 4.95, the granules, which were mainly p e r i n u c l e a r i n p o s i t i o n and f a i r l y uniform i n s i z e ( F i g . 63), s t a i n e d b r i g h t red; with i r o n hematoxylin the granules s t a i n e d b l a c k , r e a d i l y and i n t e n s e l y ( F i g . 65): with phosphotungstic  a c i d hematoxylin,  they s t a i n e d b l u e -  b l a c k , i n c o n t r a s t to the r e d s t a i n i n g of the stroma and the pale blue s t a i n i n g of the n u c l e i  ( F i g . 64).  The  granules are always most numerous and large i n s i z e i n the a c i n a r c e l l s approaching  degeneration.  In the ducts they  seem to coalesce to form l a r g e , i n t e n s e l y s t a i n i n g masses of  secretion. F o l l o w i n g the d i f f e r e n t i a l  s t a i n i n g technique of  -52-  Beaver (i960) upon the mouse p r e p u t i a l gland, only o i l r e d 0 d r o p l e t s were observed expected  throughout  the parenchyma and as  on the b a s i s of the procedures  no p e r i n u c l e a r granules were observed.  d e s c r i b e d above, However, i n the r a t  gland both l i p i d d r o p l e t s and p r o t e i n granules were demonstrated simultaneously w i t h t h i s method ( F i g . 68). W i t h i n the a c i n a r c e l l s of the r a t gland and most conspicuously, w i t h i n the c e l l s of the h y p e r t r o p h i e d a c i n i , the i n t e n s e l y b l u e - s t a i n i n g granules p r i m a r i l y surround and hence obscure  the nucleus; the o i l red 0 d r o p l e t s have a  more random d i s t r i b u t i o n .  The ducts of the gland are  f i l l e d with large o i l r e d 0 d r o p l e t s and clumps of blue s t a i n e d m a t e r i a l of considerable s i z e which must be formed by the coalescence of many p r o t e i n granules.  Unfortunately,  with Beaver's method, many d e t a i l s of the p r o t e i n granules w i t h i n the a c i n i were obscured by strong cytoplasmic and stromal d e t a i l , brought as the c o u n t e r s t a i n . the simultaneous products  about by the use of hematoxylin  For t h i s reason a second method f o r  differential  s t a i n i n g of both s e c r e t o r y  i n the r a t p r e p u t i a l gland was devised u s i n g o i l  red 0 and i r o n hematoxylin, M a t e r i a l s and Methods.  as d e s c r i b e d i n the s e c t i o n on  This second method c l e a r l y  demonstrates the d i s t r i b u t i o n of p r o t e i n granules and f a t d r o p l e t s (red) w i t h i n the gland  (black)  ( F i g . 69). Moreover,  i t documents the d e t a i l s obscured by Beaver's method -  -53-  the development of the p r o t e i n granules w i t h i n the a c i n i from t i n y granules the hypertrophied the ducts.  i n the small a c i n i to l a r g e r  spheres i n  a c i n i to masses of considerable s i z e i n  U n f o r t u n a t e l y , what was gained with one  s e c r e t o r y product was l o s t with the other; the i n t e n s i t y of, o i l red 0 s t a i n i n g was not as s a t i s f a c t o r y with second method.  this  DISCUSSION  Histology D e v e l o p m e n t a l l y and h i s t o l o g i c a l l y t h e p r e p u t i a l g l a n d s o f t h e mouse a r e e x c e s s i v e l y glands  (Voss, 1932).  as t h e s o u r c e  attractant view i s  as t h e s o u r c e  (Bronson,  1966;  Gaunt,  t h e f a v o u r e d one and w i l l  consideration later in this and S c h a f f e r preputial those  sex g l a n d s ,  emphasize  sebaceous  Both Voss  (1932) of  the  (neglecting  t h e i r r e l a t i o n s h i p to  g l a n d s of the s k i n .  the i n t i m a t e a s s o c i a t i o n  These  the  authors  of the p r e p u t i a l throughout  the  gland lifetime  animal. A t y p i c a l sebaceous  simple  further  g l a n d s i n t h e n o r m a l a d u l t male mouse  with a h a i r f o l l i c l e which p e r s i s t s of the  latter  brief descriptions  of the f e m a l e ) , r e c o g n i s e  ordinary  sexual  The  be g i v e n  discussion.  ( 1 9 3 3 ) , who g i v e  during  or s i t e of a 1967).  the  and e i t h e r  of a n a t u r a l l u b r i c a n t  c o p u l a t i o n or serve  sebaceous  F u n c t i o n a l l y , t h e y may have  c h a r a c t e r of c e r t a i n a c c e s s o r y serve  developed  gland, which i s  a  or branched h o l o c r i n e , a l v e o l a r g l a n d ,  pear-shaped, usually  d i f f e r e n t i a t e s f r o m t h e c e l l s of t h e e x t e r n a l r o o t of a growing h a i r f o l l i c l e sebaceous  gland,  thus,  duct of  opens i n t o t h e h a i r  the s t r a t i f i e d e p i t h e l i a l w i t h the outer r o o t  The s h o r t  the  c a n a l so  c e l l s of the duct are  sheath c e l l s of the h a i r  sheath  that  continuous  follicle.  The  e p i t h e l i u m of the duct  the p e r i p h e r a l l a y e r gland alveolus.  of cuboidal c e l l s  The s e b a c e o u s c e l l s  continuously arise  t o become  cytoplasmic  of  the duct The  simple later at  of  (Odland, preputial  stages  hypertrophied  surrounded  follicle  shaft  layer  as a h a i r  canal.  of cuboidal cells  as i n t h e  follicle,  which i s i n  i s already a  fully  complex, t h e r e l a t i o n s h i p  and p r e p u t i a l  g l a n d has u n d e r -  The l o n g e x c r e t o r y d u c t ,  exits,  w i t h the p r i m o r d i a l gland s t r u c t u r e s ) than  inits  However, i n c o n t r a s t t o t h e o r d i n a r y  i n emphasis.  which the h a i r  than  w i t h the c u b o i d a l  The h a i r  sebaceous g l a n d - h a i r f o l l i c l e  gone a s h i f t  the  The p r i m o r d i a l c e l l s a r e ,  by a s i n g l e  sebaceous g l a n d .  between t h e h a i r  by the  i n the r e g i o n  i n t h e n e o n a t a l mouse  of development.  structure.  fatty  g l a n d more c l o s e l y r e s e m b l e s  p r o x i m i t y t o the a c i n a r c e l l s ,  developed  The  cells  the e p i t h e l i u m of the e x c r e t o r y duct,  close  cells  i s then produced  w h i c h seems t o be c o n t i n u o u s  ordinary  the c e l l s o f  1966).  sebaceous g l a n d  t h a t time,  cells  of these  from  droplets.  s e c r e t i o n o f the sebaceous g l a n d disintegration  of the sebaceous  large spherical  lipid  with  of the a l v e o l u s  and d i f f e r e n t i a t e  the p e r i p h e r a l l a y e r engorged w i t h  i s , i n t u r n , continuous  through  i s more c l e a r l y a s s o c i a t e d  (here t h e l a r g e r  i t i s associated with Furthermore,  o f t h e two  the h a i r  follicle  the f o r m a t i o n of the  -56-  excretory duct of the p r e p u t i a l gland involves d i s t a l l y , the degeneration of hypertrophied, f a t - l a d e n acinar c e l l s and p r o x i m a l l y , the desquamation of e p i t h e l i a l  cells.  But, i n mouse f o l l i c l e s of the s k i n , sebaceous c e l l s have not been observed to take a part i n the formation of the h a i r canal which a c t u a l l y r e s u l t s only from the k e r a t i n i z a t i o n of c e l l s i n the epidermis (Hardy,  1949).  In f e t a l l i f e , the duct of the p r e p u t i a l gland i n i t s e a r l y development as a primordium, may be more d i r e c t l y r e l a t e d to the formation of the h a i r f o l l i c l e .  Unfortunately,  a study of the development of the p r e p u t i a l gland i n f e t a l mice was not undertaken f o r t h i s t h e s i s .  Such a study  would prove i n t e r e s t i n g i n determining the i n t r i c a c i e s of the a s s o c i a t i o n between the p r e p u t i a l gland and the h a i r f o l l i c l e p r i o r to b i r t h and the time of i n i t i a t i o n of these structures i n the embryo.  Regarding the l a t t e r , i t  i s known i n the mouse that the period of development f o r first-formed f o l l i c l e s ,  from the stage when the primordium  i s f i r s t seen to the stage when the h a i r emerges at the skin surface, i s nine days (Lyne, 1966).  Since the h a i r  f o l l i c l e of the p r e p u t i a l gland i s f u l l y formed i n the newborn mouse, the anlagen of the h a i r f o l l i c l e and i t s associated p r e p u t i a l gland probably appears some nine days e a r l i e r , at the end of the second week of f e t a l  life.  ¥ith the establishment of the compound a l v e o l a r pattern p r e v i o u s l y described, the g r e a t l y enlarged p r e p u t i a l gland of the adult mouse bears l i t t l e resemblance to the  -57-  simple  sebaceous g l a n d .  sebum s t i l l  results  a h o l o c r i n e type  acini,  i n which there  the  i n the  centre  of s e c r e t i o n .  i s a gradual  cells  o f the  acini  gland.  nearest  But,  i n the  my  description,  changing  of t h e i r  gland.  new  are  cells  there  preputial  gland,  acini  arise  can  a c i n i which are the  animal  depleted, gland by  the  i s no  and  exist  the  throughout acini,  lipid  the  adult  i n various the The  everbreakdown  f e a t u r e of  the  i n which  p e r i p h e r y of  r e g e n e r a t i o n of a c i n a r c e l l s has  between the  a g e s , and  larger  small  striking  age  decrease  o f the  duct  paired preputial  glands  o f the  i n b o t h male and  female r a t s ,  acini.  preputial  of a c i n a r t i s s u e  o f the mouse.  epithelioid  become  change o f the  size  i n most r e s p e c t s t o t h o s e  New  parenchymal  the p r i m o r d i a l a c i n i  the  i n the  f o r m e d sebum.  o n l y , by m i t o s i s , f r o m t h e found  of  stratification  sebaceous g l a n d ,  acinus  states  types  f r o m the  subsequent  i s a unique  the  once the  overall  of  of  (as i n the  a r e numerous t o o .  Unlike  i n c r e a s e i n the The  hypertrophied  c y c l e o f sebum f o r m a t i o n ,  acini  the most  i s the  position,  ducts  c o n s t a n t l y produced a t the  alveolus,  As  do  peripheral cells,  preputial  gland  Conglomerates  p a t t e r n of d u c t s , which r e s u l t s  of h y p e r t r o p h i e d  of  accumulation  the  the u n i f o r m  of a c t i v i t y previous  of p r e p u t i a l  p r o g r e s s i n g f r o m the p e r i p h e r y t o  sebaceous g l a n d a l v e o l u s ) , preputial  formation  from a d i s i n t e g r a t i o n  cells,  droplets  The  accompanied  system. r a t are Their on  similar  subcutaneous  e a c h s i d e of  the  -58-  midline  i n the  genital region  as  i n the  mouse.  that  difference male and equally  f e m a l e mouse, t h e  extent i n external t e n d t o be  alveolar  club.  of the  appearance; i n the  Microscopically,  f o r m a t i o n and  aging h i s t o l o g y  (Montagna and  However, a s t r i k i n g d i f f e r e n c e between the the with  association  I960).  t h e r e i s i n the Beaver  ( i 9 6 0 ) has  he  i n the  mouse) b u t  concludes that  g l a n d i n an  life  r e l a t e d to the  in hair  I960).  the  rat is gland  or e v e n a f t e r 1933;  Beaver, interstitially  extend into a duct c a p s u l e o f the  their inclusion within  area r i c h  and  because t h e s e , i f  e x t e n d b e y o n d the  " p r o b a b l y f o r t u i t o u s and  sebum  anatomy  found h a i r f o l l i c l e s  not  of  rat preputial  mouse ( S c h a f f e r ,  s e c t i o n , do  gland,  compound  Beaver,  mouse and  of the  followed (as  holocrine,  i n microscopic  o n l y r a r e l y and,  serial  glands  rat preputial  N o b a c k , 1946;  i n r a t g l a n d s , but by  certain  mouse, t h e  the  a central hair f o l l i c l e i n adult  b i r t h as  to a  adult  s i m i l a r developmental  p r e p u t i a l g l a n d s of the  a b s e n c e of any  male  a s i m i l a r cycle  d u c t f o r m a t i o n , and  as  r a t , t h e y more r e s e m b l e  mouse, e x h i b i t s t h e with  adult  almost  i n the  male r a t d i f f e r  structural pattern  size  g l a n d s of the  more f l a s k - s h a p e d ; i n t h e  a flattened that  adult  The  same  obvious  g l a n d s of the  f e m a l e as  Beaver, I960). the  the  g l a n d s of r a t s are  w e l l d e v e l o p e d i n the  male mouse and  a n i m a l , i s the  However, u n l i k e  w h i c h e x i s t s b e t w e e n the  ( B u s c h k e , 1933;  like  of t h e  the  gland i s  d e v e l o p m e n t of  follicles."  gland,  the  Physiology Since the i n i t i a l  experiments  Dennison (1934a, 1934b, 1935,  of Korenchevsky and  1936a, 1936b) on the h i s t o l o g y  of v a r i o u s sex organs of the r a t , i n c l u d i n g the p r e p u t i a l glands, a f t e r treatments with male or female s e v e r a l authors  (Salmon, 1938;  Barnes and Swyer, 1951;  sex hormones,  Noble and C o l l i p ,  1941;  Rennels, Hess, and F i n e r t y ,  Huggins, Parsons, and Jensen,  1955;  H i l f , I o v i n o , and M i c h e l , 1964) a t t e n t i o n on the responses  Beaver, 1960;  1953;  Freeman,  have a l s o centered t h e i r  of the r a t p r e p u t i a l gland to  androgens, estrogens and other hormones.  Their findings  r e p e a t e d l y show t h a t the p r e p u t i a l gland i s s t i m u l a t e d by androgens but not by estrogens.  Cursory e n d o c r i n o l o g i c a l  i n v e s t i g a t i o n s i n v o l v i n g the mouse p r e p u t i a l gland have been c a r r i e d out by Voss (1932) and Burdick and Gamon (1941), whose f i n d i n g s e s t a b l i s h the p o s i t i v e response p r e p u t i a l gland to androgens.  of the mouse  However, a d e t a i l e d  of the e f f e c t s of androgens and estrogens on the  description  overall  microscopic s t r u c t u r e of the gland i n male and female mice of d i f f e r e n t ages and of d i f f e r e n t s t a t u s (which has been presented by Beaver (i960) f o r the r a t ) i s not found i n the l i t e r a t u r e , to my knowledge. The  simple  sebaceous glands of the white r a t respond  p o s i t i v e l y to androgens and n e g a t i v e l y to estrogens. E b l i n g (1948) has found that t e s t o s t e r o n e propionate  ( l mg.  daily  -60-  f o r 36 d a y s ) i n c r e a s e s sebaceous glands,  t h e a c t i v i t y and t h e s i z e o f t h e  (the r e s u l t of hyperplasia  and h y p e r t r o p h y )  whereas e s t r a d i o l benzoate i n e i t h e r massive  (lQO u g . d a i l y  f o r 36 d a y s ) o r m o d e r a t e d o s e s ( l u g . d a i l y f o r 36 d a y s ) causes a t r o p h y of the glands.  H a s k i n , L a s h e r , a n d Rothman  (1953) have a l s o shown t h a t t e s t o s t e r o n e treatment the  propionate  ( l mg. d a i l y f o r 30 d a y s ) i n d u c e s e n l a r g e m e n t o f  sebaceous glands  Therefore,  (a 4 0 0 % average i n c r e a s e  i n size).  i t i s not s u r p r i s i n g that the p r e p u t i a l glands  r e a c t a c c o r d i n g l y when t r e a t e d w i t h t h e s e h o r m o n e s . i n t a c t a d u l t and w e a n l i n g m i c e r e c e i v i n g propionate  testosterone  ( d a i l y f o r 3 weeks), the s t i m u l a t i o n of the  female glands i s s i g n i f i c a n t l y greater gland,  In  t h a n t h a t o f t h e male  n o t o n l y on t h e b a s i s o f r e l a t i v e w e i g h t b u t a l s o  m i c r o s c o p i c a l l y , as m a n i f e s t b y h y p e r p l a s i a  of the acinar  cells,  increased  t h e h y p e r t r o p h y o f t h e a c i n i a n d by  ductular  transformation,  a male-like  a l l of which give  structural pattern.  the female  I n newborn mice, however,  male a n d f e m a l e g l a n d s r e s p o n d a l m o s t e q u a l l y t o stimulation.  gland  testosterone  The r e a s o n f o r t h e d i f f e r e n c e i n r e s p o n s i v e n e s s  among t h e a d u l t and t h e w e a n l i n g g l a n d s i s n o t c l e a r . adult glands, the  a t l e a s t , i t may be due t o t h e f a c t t h a t i n  i n t a c t a d u l t male, s t i m u l a t i o n of the gland by the  a n i m a l ' s own a n d r o g e n i c hormones i s . a l r e a d y m a x i m a l . Apparently,  Among  i n the r a t , the female glands o f a l l groups  -61-  (newborn, immature weanling, and to testosterone  a d u l t animals) respond  to a greater extent  than the male glands  (Beaver, I960) so that Beaver (i960) suggests that e f f e c t i s more l i k e l y due  to an inherent  the  sex d i f f e r e n c e  than to the f a c t that androgenic s t i m u l a t i o n of the male gland by the animal's own an inherent  hormones i s near maximal.  sex d i f f e r e n c e may  also be the  Such  important f a c t o r  f o r the d i f f e r e n c e i n responsiveness among the a d u l t  and  weanling glands of the mouse. The  f a c t that the microscopic  p r e p u t i a l gland  p a t t e r n of the  of the a d u l t male mouse i s maintained by  presence of androgens i s f u r t h e r i l l u s t r a t e d by the changes which occur i n the gland a d u l t animal and The  thus,  the  atrophic  a f t e r c a s t r a t i o n of  the  i n the absence of t e s t i c u l a r androgens.  atrophy i s manifest by a r e v e r s i o n to a non-stimulated,  somewhat f e m a l e - l i k e  p a t t e r n , which, however, as shown,  can be  counteracted  by the a d m i n i s t r a t i o n  to the  c a s t r a t e d animal.  of  testosterone  Voss (1932) too, found that  t e s t e s t r a n s p l a n t s or androgen i n j e c t i o n s to 4 week c a s t r a t e s could reverse  p r e p u t i a l glandular  reappearance of the  atrophy, b r i n g i n g about a  s e c r e t o r y a c t i v i t y of the gland.  the female mouse a f t e r castrata.cn, microscopic the  s t r u c t u r e of the p r e p u t i a l gland  gland  continues  changes i n  are n e g l i g i b l e ; the  to e x h i b i t the t y p i c a l non-stimulated  s t r u c t u r a l p a t t e r n of the a d u l t female. the mouse, there  In  i s a post  In the r a t , l i k e  c a s t r a t i o n atrophy i n the male  -62-  g l a n d but not i n the female The  gland  (Beaver,  I960).  mouse p r e p u t i a l g l a n d i s n o t s t i m u l a t e d b y  estrogens.  I n f a c t , the a c t u a l response  of the g l a n d u l a r  parenchyma t o p r o l o n g e d t r e a t m e n t s of e s t r a d i o l seems t o depend on t h e age  and  benzoate  s t a t u s of the a n i m a l ; the  g l a n d s of newborn mice are l i t t l e  a f f e c t e d ; the glands  w e a n l i n g m i c e , b o t h male and f e m a l e , show i n h i b i t e d the glands of i n t a c t a d u l t males a c i n a r a t r o p h y and  of  growth;  (but not of females)  show  an o v e r a l l d e p r e s s i o n o f s e c r e t o r y  a c t i v i t y as m a n i f e s t by d e c r e a s e d d u c t u l a r t r a n s f o r m a t i o n . The  atrophic  changes of the l a t t e r , however, a l s o  occurred  i n t h e g l a n d s o f c a s t r a t e d m a l e s and were a t t r i b u t e d t o t h e absence of the t e s t e s .  Perhaps  i t may  be  concluded,  therefore,  t h a t the a t r o p h y of the p r e p u t i a l g l a n d w h i c h f o l l o w s  the  injections  due  of e s t r a d i o l  i n t o the i n t a c t animal i s not  t o t h e s u p p r e s s i o n o f t h e g l a n d by t h e e s t r o g e n p e r b u t i s due tropic  to the e s t r o g e n i c s u p p r e s s i o n of the  e s t a b l i s h e d t h a t the a n t e r i o r which produces  organs.  The  fact  lobe of the  results  i s now  well  pituitary,  t h e g o n a d o t r o p h i c hormone c o m p l e x , FSH  LH e x e r t s a c o n t r o l l i n g gonadal  gonado-  f u n c t i o n of the a n t e r i o r p i t u i t a r y , which  i n a l o s s o f endogenous a n d r o g e n .  se  and  i n f l u e n c e upon t h e g o n a d s , t h e  s e c r e t i o n s and h e n c e , t h e a s s o c i a t e d a c c e s s o r y The  a c t i v i t y of the a n t e r i o r p i t u i t a r y ,  o t h e r h a n d , i s t o some e x t e n t c o n t r o l l e d by t h e hormones b e c a u s e when p r e s e n t i n e f f e c t i v e  on  the  gonadal  amounts,  gonadal  -63-  hormones, of e i t h e r sex,  depress the  s e c r e t i o n s of the a n t e r i o r p i t u i t a r y  gonadotrophic (apparently  indirectly,  by t h e i r a c t i o n on c e r t a i n areas of the hypothalamus that are concerned with hypophyseal a c t i v i t y ) .  Moore and  Price  (1932), among the r e s u l t s of s e v e r a l experiments, give evidence that the  i n j e c t i o n of o e s t r i n i n t o normal a d u l t 1  male r a t s leads to t e s t i c u l a r damage, a l o s s of hormone s e c r e t i o n and accessory  the  i n v o l u t i o n of the  organs (the prostate  and  the  testis  reproductive  seminal v e s i c l e s ) .  However, the i n j e c t i o n of o e s t r i n accompanied by administration  the  of f r e s h hypophyseal implants does not have  a harmful e f f e c t upon the male reproductive  system.  i n t e r p r e t a t i o n of these f i n d i n g s i s based upon the  Their complex  i n t e r r e l a t i o n s of the gonads and the a n t e r i o r p i t u i t a r y mentioned above and may  be e q u a l l y v a l i d here i n the  matter of the response of the p r e p u t i a l gland i n i n t a c t male mice.  as  The  e s t r a d i o l may  to e s t r a d i o l  so suppress  the  s e c r e t i o n of hypophyseal LH that an i n s u f f i c i e n t amount i s a v a i l a b l e f o r the endocrine f u n c t i o n of the t e s t i s .  In the  absence of t e s t i c u l a r androgens, the p r e p u t i a l glands  be-  come a t r o p h i c , i n a manner s i m i l a r to that which occurs a f t e r t e s t e s removal. complicated  However, t h i s i n t e r p r e t a t i o n i s  by the f a c t that  steroid-gonadotrophin  r e l a t i o n s h i p s do not always f a l l  i n t o the  simple  o u t l i n e d above; recent evidence suggests that the p l a y of s t e r o i d s ( p a r t i c u l a r l y . e s t r o g e n ) and  pattern inter-  gonadotrophins,  i n the female mammal, depends on the dose of estrogen  -64-  i n j e c t e d ; small doses of estrogen stimulate FSH large doses  output,  of estrogen i n h i b i t FSH output, and moderate  doses of estrogen s t i m u l a t e LH r e l e a s e (Tepperman, 1968). In my  study, the i n j e c t e d dose of estrogen was  0.00165 mg.  low  (only  d a i l y ) , but i t i s impossible to say whether  such a dosage, given during prolonged treatment, i s p h y s i o l o g i c a l l y low, moderate, or h i g h , f o r the mouse. Therefore, I am unable to suggest, w i t h any v a l i d i t y , steroid-gonadotrophin r e l a t i o n s h i p s which may  the  e x i s t i n my  experimental mice, a f t e r the i n j e c t i o n s of e s t r a d i o l . In the i n t a c t male r a t , there i s evidence small doses of estrogen produce secreting, i n t e r s t i t i a l  that  atrophy of the androgen-  c e l l s of the t e s t i s , by  directly  i n h i b i t i n g them, whereas l a r g e r doses i n the i n t a c t  animal  stimulate these same c e l l s by i n c r e a s i n g the output of hypophyseal  LH  (Tepperman, 1968).  dosage of e s t r a d i o l used i n my  Assuming that the  study i s , indeed, p h y s i o l o -  g i c a l l y low, then a r e d u c t i o n i n t e s t i c u l a r androgens, brought  about by the d i r e c t e s t r o g e n i c i n h i b i t i o n of the  Leydig c e l l s , may  account f o r the atrophy of the p r e p u t i a l  glands of the i n t a c t adult and weanling male mice. An a d d i t i o n a l h y p o t h e s i s , which cannot be e l i m i n a t e d as an e x p l a n a t i o n of the response  entirely  of the mouse  p r e p u t i a l gland to estrogen i s the p o s s i b i l i t y of a d i r e c t suppressive a c t i o n of the estrogen on the parenchyma of the p r e p u t i a l gland, i t s e l f .  Evidence from the r a t suggests  that  -65-  s u c h an e x p l a n a t i o n may  be u n l i k e l y ;  the p r e p u t i a l  glands  o f t h e f e m a l e r a t a r e a l m o s t as w e l l d e v e l o p e d as t h o s e t h e male and a f t e r p r o l o n g e d t r e a t m e n t s o f e x o g e n o u s the female  (Beaver, I960).  glands are not a f f e c t e d  e s t r o g e n , i n d e e d , a c t s d i r e c t l y and t h e p r e p u t i a l g l a n d , one w o u l d  estradiol, If  s u p p r e s s i v e l y upon  perhaps  expect the  g l a n d s o f i n j e c t e d a n i m a l s t o be a p p r o p r i a t e l y  female  affected.  However, i t i s d a n g e r o u s t o make e x t r a p o l a t i o n s f r o m sex t o t h e o t h e r s i n c e t h e r e may difference  be  i n v o l v e d i n the responses  g l a n d s o f male and f e m a l e  of  one  an i n h e r e n t s e x of the  preputial  animals to the e s t r o g e n .  F u r t h e r m o r e , because of s p e c i e s d i f f e r e n c e s i t i s o f t e n i n a d v i s a b l e t o e x t r a p o l a t e f r o m one in this  species to another,  i n s t a n c e , f r o m t h e r a t t o t h e mouse. I n a d d i t i o n t o the s u p p r e s s i v e e f f e c t s of  estradiol  upon t h e mouse p r e p u t i a l g l a n d ( e s p e c i a l l y , t h e male gland), which  I am u n a b l e  to a t t r i b u t e  t o one  particular  mechanism o f a c t i o n , t h e r e a r e o t h e r h i s t o l o g i c a l w h i c h , I t h i n k , must r e s u l t f r o m a d i r e c t a c t i o n e s t r o g e n on t h e p r e p u t i a l g l a n d .  changes of  These c h a n g e s - n a m e l y ,  t h e m e t a p l a s i a o f t h e squamous e p i t h e l i u m o f t h e main e x c r e t o r y d u c t and t h e c e n t r a l d u c t w i t h i n d i c a t i o n s t h e i r k e r a t i n i z a t i o n , and t h e d e s q u a m a t i o n cells  of  i n t o these ducts - are noted, o n l y a f t e r  of  epithelial estradiol  t r e a t m e n t , i n some g l a n d s o f b o t h s e x e s i n t h e a d u l t  and  -66-  weanling  groups.  Several authors give evidence  of s i m i l a r  d i r e c t e s t r o g e n i c e f f e c t s i n the accessory r e p r o d u c t i v e glands of the r a t and the mouse a f t e r prolonged Burrows (1935, 1937)  treatment.  d e s c r i b e s the p a t h o l o g i c a l c o n d i t i o n s  induced by large doses of e s t r o g e n i c compounds i n the c o a g u l a t i n g gland, the p r o s t a t e , and the b u l b o - u r e t h r a l gland of the male mouse; these g e n e r a l l y take the form of  e p i t h e l i a l metaplasia which, i n the p r o s t a t e and  b u l b o - u r e t h r a l gland, occurs i n the ducts, f i r s t l y , then g r a d u a l l y spreads  through  the body of the  and  gland.  In the c o a g u l a t i n g glands, which show the most marked changes, e p i t h e l i a l metaplasia i s accompanied by k e r a t i n i z a t i o n ; the sacs of the gland become f i l l e d with k e r a t i n i z e d m a t e r i a l , desquamated c e l l s and  polymorpho-  nuclear leucocytes which r e c a l l the c y c l i c a l phenomena of  e s t r u s i n the mouse v a g i n a . A r a i  (1968) found  squamous  e p i t h e l i a l metaplasia i n the seminal v e s i c l e s and c o a g u l a t i n g glands of male r a t s which was treatments  apparently caused by  of high doses of estrogen and was  neonatal  still  present 11 months a f t e r the l a s t i n j e c t i o n . The  coagulating  glands a l s o showed c o r n i f i c a t i o n l i k e those of the mouse. No d i r e c t e s t r o g e n i c e f f e c t s have been d e s c r i b e d i n the r a t gland by Beaver ( i 9 6 0 ) , a f a c t which i s perhaps r e l a t e d to the o b s e r v a t i o n that estrogen e f f e c t s on male accessory glands are u s u a l l y more marked i n the mouse than i n the r a t (Price and Williams-Ashman, 1961). Beaver  (i960)  -67-  used low doses of estrogen. high doses of estrogen  I t i s p o s s i b l e , then, that  (as A r a i , 1968,  used) and  different  periods of i n j e c t i o n would a f f e c t the r a t p r e p u t i a l  gland  directly.  P o s s i b l e endocrine p r e p u t i a l glands  r e l a t i o n s h i p s i n developing and  At 5 weeks, the  aging  compound a l v e o l a r s t r u c t u r a l p a t t e r n  of the mouse p r e p u t i a l gland i s w e l l developed, of h o l o c r i n e s e c r e t i o n and  the c y c l e  duct formation, a c t i v e ,  and  sexual d i f f e r e n c e s between male and female glands, During the i n t e r v a l from 5-8 weeks, sexual becomes very obvious  apparent.  divergence  as the male gland undergoes a tremendous  change i n s i z e and weight which i s manifest, m i c r o s c o p i c a l l y , by a s u b s t a n t i a l i n c r e a s e i n the amount of parenchyma i n the gland.  Holocrine s e c r e t i o n i s , by then, v e r y a c t i v e i n the  male gland; hence there develops  the elaborate and  p a t t e r n of ducts which i s not seen i n the female  extensive  gland.  C o i n c i d i n g w i t h t h i s development of the p r e p u t i a l gland i s the attainment  of sexual m a t u r i t y i n the animal, i t s e l f .  p r e c i s e time of sexual maturity, which depends on the balance  and  i n t e r a c t i o n of a n t e r i o r p i t u i t a r y and  hormones, i s h i g h l y v a r i a b l e i n Mus usual i n d i v i d u a l v a r i a t i o n s and  The  proper  gonadal  musculus (there are  strain variations).  the  Further-  more, i t must be i n t e r p r e t e d w i t h i n the framework of the measurement used to d e s c r i b e sexual maturation.  For example,  i n d i c e s of sexual maturity f o r the female mouse can be  the  -68-  f i r s t appearance of the v a g i n a l o r i f i c e , or the  first  appearance of the c o r n i f i e d v a g i n a l smear (both of these are estrogen dependent),  or the i n i t i a l w i l l i n g n e s s to mate, or  f i n a l l y , the a b i l i t y to conceive and c a r r y a l i t t e r fahich  to term  does not always occur with the schedule of the previous  events); f o r the male, i n d i c e s of sexual maturity can be the f i r s t appearance of spermatozoa i n the epididymis, or the i n i t i a l w i l l i n g n e s s to mate.  Because my  c h i e f concern i s  the e n d o c r i n o l o g i c a l c o n d i t i o n of the gonads, a d e f i n i t i o n of sexual m a t u r i t y as the c a p a c i t y of the gonads to f u n c t i o n , which i m p l i e s gonadal hormone a c t i v i t y but excludes reference to b e h a v i o u r a l c y c l e s , i s adequate.  any  I have not  been able to f i n d i n the l i t e r a t u r e a s p e c i f i c reference t o the age  of Swiss a l b i n o mice at normal sexual m a t u r i t y , but  i n our breeding l a b , a young female mouse of the Swiss  strain  kept w i t h males of the same l i t t e r w i l l u s u a l l y have i t s first  litter  at 8 weeks o f age.  Presumably then, sexual  maturity i s a t t a i n e d i n both sexes by the age of 5 weeks. This f i g u r e agrees w e l l w i t h the f i n d i n g s of s e v e r a l authors on the age mouse.  of m a t u r i t y i n undesignated  s t r a i n s of the a l b i n o  According to Cockrum (1962), Mus  maturity i n 35 days (that i s , 5 weeks). (1927) found the average  musculus reaches Engle and Rosasco  age of the f i r s t appearance of the  v a g i n a l o r i f i c e i n a l b i n o mice to be 35 days with a range from day 28 to day 49 whereas M i r s k a i a and Crew (1930) found the mean age of f i r s t  e s t r u s to be  39 days, with male  -69-  mice reaching maturity at about the same time.  Since  the s e c r e t i o n of androgen controls the a c t i v i t y of the p r e p u t i a l gland, i t i s not s u r p r i s i n g that the maturation of the male gland coincides with the age of sexual maturity at about 5 weeks and i s followed by the establishment of the stimulated pattern during the i n t e r v a l from 5-8 weeks.  At t h i s time androgen  production i n the male mouse must be very a c t i v e . In a d d i t i o n to the postpuberal growth of the male p r e p u t i a l gland there i s a prepuberal period of steady growth by weight (see graph l ) which i s manifest m i c r o s c o p i c a l l y by increases i n the amount of glandular parenchyma.  In the r a t , i t i s known that the secretory  a c t i v i t y of the testes gradually increases during prepuberal development, i n response to the r i s i n g t i t e r s of c i r c u l a t i n g gonadotrophins, u n t i l the l e v e l of androgen maintained i n the adult i s reached (Moore and P r i c e , 1938).  S i m i l a r secretory a c t i v i t y by the a n t e r i o r  p i t u i t a r y and the developing testes i n the mouse would account f o r t h i s prepuberal stage of p r e p u t i a l gland growth.  There i s evidence to support t h i s  assumption.  For example, i t i s known that the d i f f e r e n t i a t i o n of the mouse hypophysis from a gonadotrophic viewpoint occurs at day 6 i n males and at day 12 i n females  (Bronson,  Dagg, and S n e l l , 1966) and that t h e r e a f t e r the capacity of the gonads to respond to gonadotrophic hormone s t i m u l a t i o n  -70-  develops g r a d u a l l y , i n c r e a s i n g r a p i d l y at about 3 weeks of age  ( P r i c e and O r t i z , 1944).  Therefore, i t i s  reasonable to assume that the p r o d u c t i o n of t e s t i c u l a r androgens i n c r e a s e s g r a d u a l l y d u r i n g the prepuberal development of the mouse, i n response to the i n c r e a s i n g gonadotrophic a c t i v i t y .  The p r e p u t i a l gland develops  a c c o r d i n g l y ; i t s s t r i k i n g r e a c t i v i t y begins a f t e r 3 weeks of age  (see graph l ) ,  the time, at which,  i t is  i n t e r e s t i n g to note, that gonadal a c t i v i t y increases significantly. But what of androgen p r o d u c t i o n i n the female mouse?  Ye have seen that the female p r e p u t i a l gland,  l i k e that of the male, i s s t i m u l a t e d by androgens but not by estrogens. unique  The female p r e p u t i a l gland i s not  i n this respect.  female mouse responds too.  The p r o s t a t e homologue i n the  only to androgenic  stimulation  In the female mammal the p r o d u c t i o n of androgens  seems to occur normally i n the o v a r i e s and the adrenal glands (Parkes, 1950;  P r i c e and Williams-Ashman,  under the i n f l u e n c e of hypophyseal hormones.  1961),  Such  androgenic p r o d u c t i o n i n the young female mouse, i n c r e a s i n g g r a d u a l l y i n response to i n c r e a s i n g gonadotrophic a c t i v i t y , u n t i l the l e v e l of androgen maintained i n the a d u l t i s reached, would account f o r the prepuberal p e r i o d of growth a,n& development of the female p r e p u t i a l gland  -71-  (see graph 2), up to the time  of sexual m a t u r i t y when  the mature microscopic p a t t e r n of the gland i s reached and t h e r e a f t e r maintained.  Androgen p r o d u c t i o n i n the  female mouse must, of course, be very much l e s s than i n the male mouse.  Not only i s the prepuberal growth of the  female gland l e s s s t r i k i n g than t h a t of the male but i t s postpuberal growth i s almost n e g l i g i b l e , i n c o n t r a s t to t h a t of the male gland. I t i s p o s s i b l e t h a t the mouse p r e p u t i a l  gland  responds d i r e c t l y to c i r c u l a t i n g p i t u i t a r y hormones as w e l l as to androgens.  I t has been demonstrated, f o r  example, that the r a t p r e p u t i a l gland undergoes marked atrophy f o l l o w i n g hypophysectomy (Noble and C o l l i p , Beaver, I960); an atrophy which i s not  1941;  completely  r e v e r s i b l e by the a d m i n i s t r a t i o n of t e s t o s t e r o n e alone. The presence  of the p i t u i t a r y gland seems to be  f o r t e s t o s t e r o n e to exert i f s f u l l gland.  necessary  a c t i o n on the p r e p u t i a l  As a d d i t i o n a l evidence, Noble and C o l l i p  (l94l)  have shown i n female r a t s that p i t u i t a r y e x t r a c t s can stimulate the p r e p u t i a l glands to a c e r t a i n extent i n the absence of o v a r i e s and adrenal glands.  The s t i m u l a t o r y  e f f e c t s of the p i t u i t a r y e x t r a c t s were augmented i n i n t a c t animals.  Perhaps then, i t i s t h i s a c t i v i t y of the  p i t u i t a r y which e x p l a i n s why  ovariectomy  does not  affect  the p r e p u t i a l gland of the a d u l t female mouse; i t i s s u f f i c i e n t to m a i n t a i n the  "non-stimulated"  p a t t e r n of the  -72-  female gland i n the absence of o v a r i a n androgens. adrenal androgens may  The  a l s o make a c o n t r i b u t i o n to the  maintenance of the p r e p u t i a l gland f o l l o w i n g ovariectomy. I t i s d i f f i c u l t to assess t h e i r r e l a t i v e  importance because  I c a r r i e d out no adrenalectomy experiments.  However, there  are i n d i c a t i o n s i n the l i t e r a t u r e t h a t adrenal androgens can u s u a l l y cause only s l i g h t e f f e c t s  ( P r i c e and W i l l i a m s -  Ashman, 1961) and i t has been shown i n female r a t s that the removal of both ovaries and adrenals does not a f f e c t the weight of the p r e p u t i a l glands (Noble and C o l l i p , I f the p r e p u t i a l gland f o l l o w s the t y p i c a l  1941).  pattern  of development of c e r t a i n mammalian r e p r o d u c t i v e accessory glands, such as the p r o s t a t e  ( P r i c e , 1936; Burns, 1961),  i t s e a r l y development i n the male mouse before b i r t h  may  be dependent on the s e c r e t i o n of male hormone by the f e t a l t e s t i s , whereas, i n the female mouse, i t s e a r l y development may proceed independently of hormonal c o n d i t i o n i n g a c e r t a i n stage),  (up to  But, since the p r e p u t i a l glands of  newborn mice, of both sexes, are so s i m i l a r i n s i z e and i n microscopic s t r u c t u r e , i t i s p o s s i b l e that the f e t a l development of male glands may be independent of hormonal s t i m u l a t i o n , as w e l l as t h a t of female glands, u n t i l the d i f f e r e n t i a t i o n of the hypophysis from the gonadotrophic p o i n t of view, at day 6 i n males and at day 12 i n females. (It i s i n t e r e s t i n g to note that t h i s i n i t i a t i o n of gonadotrophic a c t i v i t y i s followed c l o s e l y by the growth  -73of the male gland a t week 1 and of the female week 2.)  In order to determine  gland a t  the p r e c i s e r o l e of hormones  i n the e a r l y development of the male p r e p u t i a l gland, a study of the e f f e c t s of e a r l y c a s t r a t i o n on the glands of f e t a l and neonatal mice would have to be  undertaken.  The p r e p u t i a l glands of mice age q u i c k l y .  During  the i n t e r v a l from 3-12 months both male and female  glands  undergo a tremendous r e d u c t i o n and degeneration of a c i n a r t i s s u e , accompanied by an i n c r e a s e i n the s i z e of the duct system.  By 12 months, the glands are made up almost  e n t i r e l y of a network of empty ducts.  In the p r o s t a t e  glands of aging r a t s and mice (the exact age i s not s t a t e d ) , s e n i l e changes, s i m i l a r to those brought  about by c a s t r a t i o n ,  have been d e s c r i b e d by Moore (1936) and are i n t e r p r e t e d on the b a s i s of a decrease androgen.  or absence of t e s t i c u l a r  However, hormonal d e f i c i e n c i e s of the gonads are  probably not a notable f a c t o r i n the aging processes of the p r e p u t i a l gland.  These age changes are not s i m i l a r to  the a t r o p h i c changes caused by c a s t r a t i o n .  Moreover,  during the i n t e r v a l from 3-12 months, the r e p r o d u c t i v e a c t i v i t i e s of the mouse normally continue; t h i s i m p l i e s gonadal  activity.  According to S n e l l  (1941), the u s e f u l  breeding p e r i o d of the i n b r e d female mouse terminates a t about 1 year of age; t h e r e a f t e r , there i s a decrease i n f e r t i l i t y and f e c u n d i t y which i s s t r a i n dependent and hence, h i g h l y v a r i a b l e .  A c t u a l l y , a gradual decrease i n  -74-  fertility  i n aging female mice need not n e c e s s a r i l y  r e f l e c t a lowered Endocrine  estrogen p r o d u c t i o n by the o v a r i e s .  a c t i v i t y can apparently be r e t a i n e d longer  than  r e p r o d u c t i v e c a p a c i t y i n some mice, as shown by Thung, Boot and Miihlbock (1956) f o r the i n b r e d s t r a i n s C57BL, O20, the h y b r i d (O20  x  DBAf).  and  U n f o r t u n a t e l y , to my knowledge,  there has been no c o n s i d e r a t i o n of t h i s r e l a t i o n s h i p i n the Swiss s t r a i n of mice.  Still,  f o r a l l these  given above, i t seems u n l i k e l y that endocrine  reasons  deficiencies  are r e s p o n s i b l e f o r the aging of mouse p r e p u t i a l during the i n t e r v a l from 3 to 12 months.  More  glands  important  f a c t o r s i n aging must be changes i n the response  of the  p r e p u t i a l gland to androgens, once the gland has  fully  developed continue  (at 3 months i n some animals).  Androgens must  to stimulate the s e c r e t o r y a c t i v i t y of the gland  but must cease to a p p r e c i a b l y a f f e c t the r e g e n e r a t i o n of parenchyma i n the a d u l t gland. of  With the gradual d e p l e t i o n  p r i m o r d i a l a c i n i and the c e s s a t i o n of m i t o t i c  i n the a c i n a r c e l l s , the e f f e c t i v e  "life"  of the  activity preputial  gland i s f i x e d by the amount of parenchyma which makes up the gland at t h i s stage of development.  T h e r e a f t e r , the  parenchyma of the gland i n e v i t a b l y diminishes; the of  result  the c e l l u l a r d e s t r u c t i o n which, of n e c e s s i t y , accompanies  holocrine secretion.  In t h i s way,  the p r e p u t i a l gland i s  unique among the accessory r e p r o d u c t i v e glands of the  75-  mouse; f o r , i n most accessory reproductive  glands  s e c r e t o r y a c t i v i t y i s a continuous  dependent upon  an androgen stimulated  process  epithelium.  Histochemistry The h i s t o c h e m i c a l s t u d i e s of my h i s t o l o g i c a l observations  t h e s i s confirm  the  on the c y c l e of sebum formation  i n the mouse p r e p u t i a l gland; t h a t , i n f a c t , the s e c r e t i o n of the gland i s l i p i d which i s e l a b o r a t e d i n the c e l l s of the a c i n i and discharged degenerate.  The  i n t o the ducts when the  cells  l a t t e r could not be observed i n the  h i s t o l o g i c a l p r e p a r a t i o n s because of the use  of the f a t  s o l v e n t s i n the p a r a f f i n p r o c e s s i n g which d i s s o l v e d the lipid;  t h e r e f o r e , the ducts always appeared empty and  acinar c e l l s , vacuolated.  Much of the l i p i d  the  s e c r e t i o n of  the mouse p r e p u t i a l gland seems to be made up of n e u t r a l fats  ( t r i g l y c e r i d e s ) as shown by o i l red 0 s t a i n i n g .  c l a s s e s of l i p i d which are p o s s i b l y present  Other  i n the s e c r e t i o n ,  i n a d d i t i o n to the n e u t r a l f a t s , could not be  identified  by the methods of the present i n v e s t i g a t i o n . However, Gaunt (1967), using a s i l i c i c  a c i d column, separated  free f a t t y  acids as w e l l as n e u t r a l l i p i d s from mouse p r e p u t i a l gland s e c r e t i o n and P a t t e r s o n p r e p u t i a l glands  (i960) found t h a t the  of young male mice, when incubated  in  14 a c e t a t e , i n c o r p o r a t e d the C  a c t i v e l y i n f o s t e r o l s so that  -76-  these, too, may The  normally be present i n the  l i p i d s of the  examined by G a l l o  lipid  secretion.  r a t p r e p u t i a l gland have been  (1966) whose f i n d i n g s demonstrate  the  f o l l o w i n g d i f f e r e n t c l a s s e s : squalene, t r i g l y c e r i d e s , s t e r o l e s t e r s , monoglyderides, d i g l y d e r i d e s , and  free s t e r o l s .  major component.  free f a t t y acids,  Of these, squalene seems to be In c o n t r a s t ,  squalene has  not  the  been  l o c a l i z e d i n the mouse p r e p u t i a l gland, whose f a t t y s e c r e t i o n is,  therefore,  more c l o s e l y r e l a t e d to that of the  sebaceous glands of mice and replaced The  by A  - cholestenol  skin  r a t s , i n which squalene i s (Brooks, Lalic.h, Baumann, 1956).  r a t p r e p u t i a l gland, however, i n i t s p r e f e r e n t i a l  accumulation of squalene, resembles the  functional  sebaceous  glands of the human dermis i n which squalene i s also produced i n large amounts (Kandutsch, 1964). No  s p e c i f i c proteins  were l o c a l i z e d i n the  c e l l s of the mouse p r e p u t i a l gland. s t r i k i n g difference  i n a d d i t i o n to the  mouse and  l i p i d s , the  p r o t e i n granules which are The  granules have no  sebaceous gland nor  Thus, there e x i s t s a  i n the h i s t o c h e m i s t r y  p r e p u t i a l glands of the  In the  latter,  c e l l s c o n t a i n numerous  sepreted along with the  counterpart i n the i n the  between the  the r a t .  acinar  acinar  lipid.  c e l l s of the  simple  c e l l s of the mouse p r e p u t i a l  gland i n which a l l attempts to demonstrate them, f o l l o w i n g the methods of Beaver (1959, 1960), proved  fruitless.  -77-  Two  functions  gland: l ) that the l u b r i c a n t during  have been proposed f o r the  gland serves as the  copulation  a t t r a c t members of the research  i n d i c a t e that the  the.more l i k e l y one.  source of a n a t u r a l  or 2) that the  opposite sex.  preputial  The  f i n d i n g s of  latter function  Bronson (1966) has  gland serves to recent  i s probably  demonstrated  the  a t t r a c t i o n of C57BL./6J female mice to the p r e p u t i a l gland s e c r e t i o n of the male.  Gaunt (1967), a f t e r v e r i f i c a t i o n  t h i s a t t r a c t i o n i n Swiss mice, has  of  observed that i t i s the  f r e e f a t t y a c i d component of the male p r e p u t i a l gland secretion  (and not  the  neutral l i p i d  a t t r a c t s female mice and  component) which  i n addition, accelerates  of grouped females (the Whitten e f f e c t ) .  the  Although the  f a t t y a c i d component of the p r e p u t i a l gland i s the of an a t t r a c t a n t or pheromone, the pheromone and  determined.  free  site  o r i g i n a l source of  i t s i d e n t i t y remain to be  estrus  the  -78-  SUMMARY AND  The  preputial  glands  CONCLUSIONS  o f the male and  female  mouse  are p a i r e d , subcutaneous organs s i t u a t e d i n the  genital  r e g i o n o f t h e a n i m a l , one  ventral  midline. urethral  E a c h g l a n d opens o n t o  the  s i d e of t h e s k i n , beside  the  o p e n i n g , v i a an e x c r e t o r y d u c t .  The developed  on e i t h e r  p r e p u t i a l glands are b a s i c a l l y e x c e s s i v e l y s e b a c e o u s g l a n d s and  so d e v e l o p m e n t a l l y ,  like  the s i m p l e sebaceous g l a n d s , t h e y are d e r i v a t i v e s of skin.  Furthermore,  with a hair f o l l i c l e of  the a n i m a l .  each g l a n d i s i n t i m a t e l y which  p e r s i s t s throughout  the  associated the  lifetime  H i s t o l o g i c a l l y , the g l a n d s are h o l o c r i n e  compound, a l v e o l a r g l a n d s .  During p o s t n a t a l development  b o t h male and  i n c r e a s e i n s i z e , as t h e  of  female  glands  t h e p r o l i f e r a t i o n o f a c i n a r c e l l s , b u t t h e male  u n d e r g o e s the more s t r i k i n g e n l a r g e m e n t . has  result  gland  Once e a c h a c i n u s  f o r m e d sebum, t h e r e i s no r e g e n e r a t i o n o f  acini.  T h e r e f o r e , w i t h a g i n g , as t h e c y c l e of h o l o c r i n e s e c r e t i o n and  d u c t f o r m a t i o n c o n t i n u e s , t h e parenchyma o f t h e a d u l t  preputial  gland g r a d u a l l y decreases.  L i p i d s , p r i m a r i l y i n the form f a t s , which are,  accumulate  of d r o p l e t s of n e u t r a l  i n the c y t o p l a s m  o f the  acinar  t o my k n o w l e d g e , the o n l y s e c r e t o r y p r o d u c t  mouse p r e p u t i a l g l a n d .  The  lipid  of  i s discharged into  cells,  the the  -79-  ducts  of t h e g l a n d as t h e a c i n a r c e l l s The  p r e p u t i a l gland  testosterone propionate  degenerate.  of t h e mouse i s s t i m u l a t e d by  b u t n o t by  e s t r a d i o l benzoate.  The  s t i m u l a t o r y e f f e c t o f t e s t o s t e r o n e i s m a n i f e s t , most o f t e n , by h y p e r p l a s i a of t h e a c i n a r c e l l s ,  hypertrophy  acini,  The  and  by  increased secretion.  depends upon t h e age gland tends  and  to respond  of  degree of  sex of the a n i m a l .  The  the response  female  t o exqgenous t e s t o s t e r o n e t o a  g r e a t e r e x t e n t t h a n t h e male g l a n d , w i t h t h e p o s s i b l e e x c e p t i o n of t h e g l a n d s changes w h i c h o c c u r  o f newborn a n i m a l s .  i n the female  The  gland a f t e r  structural  testosterone  s t i m u l a t i o n g i v e the gland a m a l e - l i k e s t r u c t u r a l p a t t e r n . Since the androgen-stimulated f r o m the n o n ^ s t i m u l a t e d  gland d i f f e r s  g l a n d , and  p r o d u c t i o n must be v e r y much l e s s t h a t i n the male, i t i s not differences mice.  i n the glands  Furthermore,  since normally i n the female  s u r p r i s i n g to f i n d  mouse  than  morphological  o f n o r m a l a d u l t male and  by t h e p r e s e n c e  i s a p o s t - c a s t r a t i o n atrophy  female  of a n d r o g e n s ,  I n the female  the there  i n t h e male g l a n d w h i c h i s  m a n i f e s t by a r e v e r s i o n t o a n o n - s t i m u l a t e d  microscopic  androgen  b e c a u s e t h e m i c r o s c o p i c p a t t e r n of  male g l a n d i s m a i n t a i n e d  like pattern.  structurally  mouse, a f t e r  somewhat  female-  castration,  changes i n s t r u c t u r e o f tb.e p r e p u t i a l g l a n d  are  negligible. Prolonged p r e p u t i a l gland  treatments  of e s t r a d i o l benzoate a f f e c t  i n d i r e c t l y and/or d i r e c t l y .  the  Some b r i n g a b o u t  -80-  a c i n a r a t r o p h y and  a d e p r e s s i o n of s e c r e t o r y a c t i v i t y  m a n i f e s t by d e c r e a s e d  ductular transformation.  t h e a c t u a l d e g r e e of r e s p o n s e depends upon t h e age of  and  inhibited  s t a t u s of t h e a n i m a l ;  o f w e a n l i n g m i c e , b o t h male and growth;  However,  of t h e g l a n d u l a r parenchyma  i n t a c t a d u l t males (but not of f e m a l e s )  the glands  the glands A direct  as  the  glands  become a t r o p h i c ; female,  show  o f newborn mice seem t o  a c t i o n o f e s t r o g e n on t h e  be  little  affected.  pre-  putial  g l a n d i s m a n i f e s t , i n some g l a n d s , by m e t a p l a s i a  t h e squamous e p i t h e l i u m of t h e  main e x c r e t o r y d u c t , and  the desquamation of e p i t h e l i a l  cells  i n t o these  W i t h r e f e r e n c e t o the hormone t r e a t m e n t s  of by  ducts.  of t h i s  study  (with  e i t h e r t e s t o s t e r o n e p r o p i o n a t e or e s t r a d i o l b e n z o a t e ) i t must be  remembered t h a t t h e r e s u l t s  c e r t a i n doses, or  different  so t h a t i t i s p o s s i b l e t h a t d i f f e r e n t  intervals  results.  of t r e a t m e n t would g i v e  of  physiologically  ( i n terms  of  i n many r e s p e c t s t o t h o s e  organs which any  i n t h e h i s t o c h e m i s t r y of t h e s e  are i n t e r e s t i n g to note:  firstly,  the presence  the  a f t e r b i r t h as t h e r e i s i n t h e mouse; of p e r i n u c l e a r , p r o t e i n a c e o u s  the micro-  homologous  a s s o c i a t i o n of t h e r a t p r e p u t i a l g l a n d w i t h a  hair f o l l i c l e  glands  of  However, t h e r e a r e s t r i k i n g d i f f e r e n c e s i n t h e  s c o p i c anatomy and  of  different  t o hormone t r e a t m e n t s ) t h e p a i r e d p r e p u t i a l  t h e mouse a r e s i m i l a r  rat.  doses  .  H i s t o l o g i c a l l y and response  were o b t a i n e d w i t h  absence central secondly,  granules i n the  -81-  acinar  cells  counterpart  of t h e r a t p r e p u t i a l g l a n d , w h i c h have i n the a c i n a r  no  c e l l s o f t h e mouse p r e p u t i a l  gland. Developmentally,  histologically,  the p r e p u t i a l gland  o f t h e mouse r e m a i n s  developed sebaceous  gland.  Because  1960),  (Schaffer,  i n order to d i s t i n g u i s h  preputial  gland.  at least  of  histo-  secrete  t h e r a t p r e p u t i a l g l a n d has  a " d i c r i n e " gland  v a r i e t y of h o l o c r i n e  excessively  n  of i f s a p p a r e n t a b i l i t y t o  two d i f f e r e n t s u b s t a n c e s , designated  a  But the p r e p u t i a l gland  t h e r a t c a n n o t be c a t e g o r i z e d so s i m p l y , chemically.  and h i s t o c h e m i c a l l y ,  1933;  i t from the  gland to which belongs  Beaver,  "monocrine" t h e mouse  been  -82-  PLATE  1 - The Young A d u l t P r e p u t i a l G l a n d o f Mus  Fig.  1.  P r e p u t i a l g l a n d o f t h e male mouse, weeks,  showing  formation. a)  c e n t r a l duct  b)  lateral  d) 2.  a c t i v e s e c r e t i o n and  parenchyma  connective tissue  P r e p u t i a l gland  capsule  of the f e m a l e , a t 5 weeks. 1.  i n s i z j e and t h e r e l a t i v e of the d u c t system. 3.  acini,  showing  pheral  cells  present  37 X. of type 2  a l a y e r of f l a t t e n e d  ( b l a c k arrow)  of each a c i n u s .  The  476  A type 3 acinus hypertrophied,  toward the  vacuoles fatty  X. (black arrow)  composed  vacuolated c e l l s ,  '  .  of  surrounded  by a l a y e r o f f l a t t e n e d p e r i p h e r a l 476 X .  peri-  surrounding  i n some c e l l s , r e p r e s e n t  globules. 4.  underdevelopment  of i n c r e a s i n g magnitude  centre  Fig.  Note the d i f f e r e n c e  G l a n d u l a r parenchyma - d e t a i l  cells  duct  duct  Compare w i t h f i g u r e  Fig.  at 5  37 X.  c) g l a n d u l a r  Fig.  musculus  cells.  -83-  PLATE 2 - The Y o u n g A d u l t P r e p u t i a l G l a n d o f Mus Pig.  5.  A type 4 acinus beginning  whose c e n t r a l c e l l s  to degenerate  the a c i n u s . peripheral  6.  (a)  c e l l s are becoming  7.  476  o f a 5 week  and d e g e n e r a t i n g  the p e r i p h e r y  to the c e n t r e of the 146  follicle  of  a progression  hypertrophied  (b).  acini of  cells  from  acini,  X.  (a) w h i c h i s  usually  a s s o c i a t e d w i t h each p r e p u t i a l g l a n d . shaft  projects  i n t o the p o s t e r i o r  the c e n t r a l duct  (b)  near  the main e x c r e t o r y d u c t 91 Fig.  8.  The  stratified  conglomerates  i n which there i s  The h a i r  of  X.  showing  near a duct Fig.  i n the centre  D e t a i l of the p r e p u t i a l gland male mouse,  are  The n u c l e i a r e p y k n o t i c .  (black arrow). Fig.  musculus  its  (c)  as  end  union  Its of  with  shown h e r e .  X.  D e t a i l of the p r e p u t i a l g l a n d f e m a l e mouse.  Compare w i t h f i g u r e  the t h i c k e n e d f i b r o u s (stroma)  o f a 5 week  interstitial  which surrounds the a c i n i  female gland.  228  X.  6.  Note  tissue in  this  -84-  PLATE 3 - The D e v e l o p m e n t Fig.  9.  of the P r e p u t i a l  The p r e p u t i a l g l a n d (a)  Fig.  10.  duct. 11.  The h a i r  follicle Its  (black arrow)  12.  main  outer root  sheath  w i t h the  p e r i p h e r a l c e l l s of  s e c t i o n of  the h a i r  o f t h e newborn p r e p u t i a l g l a n d , shaft  cells  the  X.  Longitudinal  hair  excretory  o f t h e newborn p r e p u t i a l .  are continuous  cuboidal-shaped 228  its  gland,  X.  (a).  gland.  tissue  X.  the d e r i v a t i o n of 228  gland  Fig.  91  connective  D e t a i l o f t h e newborn p r e p u t i a l showing  Fig.  o f t h e newborn mouse  i n the s u r r o u n d i n g  of the p r e p u c e .  Gland  (a)  emerging  of the e x c r e t o r y duct  follicle showing  its  i n t o the d i s t a l  end  (b).  .228  X.  -85-  PLATE 4 - The D e v e l o p m e n t  Pig.  13«  of the P r e p u t i a l Gland  The p r e p u t i a l g l a n d of t h e m a l e mouse, 1 week o f a g e .  Fig.  14.  91 X .  The p r e p u t i a l g l a n d o f t h e male mouse, 2 weeks  of age.  The a c i n i a r e  Fig.  15.  stage  of development.  shaped  91 X .  The m a i n e x c r e t o r y d u c t o f t h e 1 week preputial  gland  (male),  showing  at  densely-  p a c k e d t o g e t h e r , and more u n i f o r m l y at t h i s  at  old  its  k e r a t i n i z e d s t r a t i f i e d squampus e p i t h e l i u m . 106  X.  1 4  -86-  PLATE 5 - The Aging of the P r e p u t i a l Gland F i g . 16.  The p r e p u t i a l gland of the male at 8 weeks. The duct system has become more extensive by ductular transformation.  F i g . 17.  The p r e p u t i a l glancl of the female, at 8 weeks.  F i g . 18.  13 X.  37 X.  The p r e p u t i a l gland of the male at 5 months, showing the s t r i k i n g decrease i n acinar t i s s u e , accompanied by a compensatory i n the duct system.  F i g . 19.  increase  11 X.  The p r e p u t i a l gland of the female at 5 months.  Note the decrease i n acinar t i s s u e ,  the abundant stroma.  37 X.  -87-  PLATE 6 - The A g i n g o f t h e P r e p u t i a l G l a n d  Fig.  20.  The p r e p u t i a l g l a n d o f t h e m a l e , a t months.  Fig.  21.  The g l a n d i s  cystic.  13 X .  The p r e p u t i a l g l a n d o f t h e f e m a l e , a t months.  increased considerably. 22.  11  The p a r e n c h y m a i s much r e d u c e d  and t h e lumen o f t h e c e n t r a l d u c t  Fig.  11  has  37 X .  D e t a i l o f t h e 11 mpnth o l d c y s t i c p r e p u t i a l gland of the male.  The c e n t r a l d u c t  f i l l e d w i t h a substance  rich in  is  lymphocytes,  polymorphonuclear n e u t r o p h i l e s  and  desquamated e p i t h e l i a l  O n l y a few  isolated acini  (a)  of the g l a n d .  37 X .  cells.  remain at the  periphery  -88-  PLATE 7 - The E x p e r i m e n t a l G l a n d s Pig.  Fig.  Fig.  23.  24.  25.  o f Group I  The p r e p u t i a l g l a n d  o f t h e male mouse  7 weeks,  showing  the normal,  compound  alveolar pattern.  12  male.  The g l a n d  atrophic.  figure  23,  12  is  developed,  castrated  Compare  The p r e p u t i a l g l a n d  o f t h e 10 week  castrated  male, a f t e r the d a i l y a d m i n i s t r a t i o n  of  e s t r a d i o l benzoate  gland  f o r 3 weeks.  gland  in figure  The p r e p u t i a l g l a n d  The  o f a t r o p h y as 24.  testosterone  propionate  testosterone  t r e a t m e n t has  m a i n t a i n i n g the a d u l t Compare w i t h f i g u r e s The p r e p u t i a l g l a n d  which i s 12 X.  showing  the r e s u l t  the  castrated of  f o r 3 weeks.  The  counteracted  of t h e g l a n d ,  the  thereby  structural pattern. 23 and 24.  of a n o t h e r  c a s t r a t e d male, a f t e r the treatment,  does  12 X.  o f t h e 10 week  p o s t - c a s t r a t i o n atrophy  27.  with  X.  male, a f t e r the d a i l y a d m i n i s t r a t i o n  Fig.  at  X.  o f t h e 10 week  control 26,  fully  The p r e p u t i a l g l a n d  shows a s i m i l a r d e g r e e  Fig.  (Males)  12 X. 10 week  testosterone  the " e x h a u s t i o n of hormonal  atrophy"  overstimulation.  -89-  PLATE 8 - The E x p e r i m e n t a l G l a n d s Fig.  28.  of Group I ( M a l e s )  D e t a i l of the p r e p u t i a l g l a n d castrate.  Note the abundant  o f t h e 10 week fibrous  a c i n a r and s u b e p i t h e l i a l s t r o m a , underdeveloped 146 Fig.  29.  acini.  prolonged  testosterone  and p o l y m o r p h o n u c l e a r the d u c t s ,  30.  small, 6.  X.  t h e 10 week c a s t r a t e d g l a n d  Fig.  and t h e  Compare w i t h f i g u r e  D e t a i l of the p r e p u t i a l gland  146  inter-  in figure  27  -  o v e r s t i m u l a t e d by-  treatments.  Lymphocytes  n e u t r o p h i l s (a)  a c i n i and s t r o m a o f t h e  invade  gland.  X.  D e t a i l o f t h e e s t r a d i o l - t r e a t e d , 10 week c a s t r a t e d p r e p u t i a l gland  (male),  showing  the d i r e c t e f f e c t s of the e s t r o g e n i c  hormone  the t h i c k e n e d s t r a t i f i e d e p i t h e l i u m of c e n t r a l d u c t , and t h e d e s q u a m a t e d cells  i n the d u c t .  146  X.  the  epithelial  -  -90-  PLATE 9 - The E x p e r i m e n t a l G l a n d s o f Group I  Fig.  31.  (Females)  The p r e p u t i a l g l a n d o f t h e f e m a l e mouse, a t 7 w e e k s . 41 X .  Fig.  32.  The p r e p u t i a l g l a n d o f t h e 10 week c a s t r a t e d female.  The g l a n d i s n o t a t r o p h i c l i k e  o f t h e c a s t r a t e d male ( f i g u r e  24).  decrease  i n t h e amount o f p a r e n c h y m a , t h e  increase  example 33.  Only-  aging e f f e c t s are observed - the  i n t h e lumen o f t h e c e n t r a l d u c t ,  Fig.  that  for  41 X .  The p r e p u t i a l g l a n d o f t h e 10 week c a s t r a t e d female, a f t e r prolonged e s t r a d i o l benzoate treatments.  Fig.  34.  Detail effects  41 X .  of f i g u r e 33, showing the d i r e c t o f t h e e s t r o g e n i c hormone.  122  X.  -91-  PLATE 10 - The E x p e r i m e n t a l G l a n d s Fig.  35.  The  p r e p u t i a l gland  of G r o u p I  (Females)  o f t h e 10 week  female a f t e r prolonged  testosterone  showing the s t i m u l a t o r y  hypertrophy  ductular 32. Fig.  36.  41  treatment,  e f f e c t of the  as m a n i f e s t b y marked h y p e r p l a s i a cells,  castrated  of the  o f t h e a c i n i and  transformation.  acinar  increased  Compare w i t h  figure  X.  D e t a i l of f i g u r e  35 - t h e s t i m u l a t e d  female  gland.  Note the h y p e r t r o p h i e d a c i n i ,  absence  of abundant  of w h i c h g i v e pattern.  hormone,  the  i n t e r a c i n a r stroma,  the gland a m a l e - l i k e  Compare w i t h f i g u r e s  all  structural  6 and 4 2 .  122  X.  -92-  PLATE  11 - The E x p e r i m e n t a l G l a n d s Adult Mice  Fig.  37.  ma,le mouse.  Fig.  38.  The  o f t h e 10 week  24.  p r e p u t i a l gland  12  39.  The p r e p u t i a l g l a n d  40.  D e t a i l of f i g u r e acini  39.  12  estradiol  the s t r i k i n g a t r o p h y  Compare w i t h f i g u r e  37.  37.  X. intact  treatment,  of the  gland.  12 X.  Note the  small-size  and t h e r a t h e r more a b u n d a n t  interacinar  hormone  transform-  o f t h e 10 week  male, a f t e r a prolonged  Fig.  treatment,  the s t i m u l a t o r y e f f e c t of the  Compare w i t h f i g u r e  showing  intact  testosterone  as m a n i f e s t b y i n c r e a s e d d u c t u l a r  Fig.  castrate  X.  o f t h e 10 week  male, a f t e r prolonged  ation.  intact  Compare w i t h t h e 10 week  in figure  showing  -Intact  (Males)  The p r e p u t i a l g l a n d  gland  of Group I i  fibrous  and s u b e p i t h e l i a l s t r o m a .  122  X.  PLATE  12 -  The E x p e r i m e n t a l G l a n d s A d u l t Mice  Pig.  41 .  42.  The p r e p u t i a l g l a n d 41  of the i n t a c t female  41.  parenchyma,  in  - the decrease the abundant  Fig.  43.  ment,  i n t h e amount the  tissue  o f t h e 10 week  a f t e r prolonged  showing  t h e hormone cells,  normal  the s t i m u l a t o r y  - the h y p e r p l a s i a  the hypertrophy  with figure 44.  invasion  elements  41.  41  The p r e p u t i a l g l a n d  intact  testosterone  treat-  effects of the  of acinar  of t h e a c i n i , and  increased ductular transformation.  Fig.  of  X.  The p r e p u t i a l g l a n d female,  the  stroma,  o f some a c i n i by c o n n e c t i v e 122  at  Note the t y p i c a l  e f f e c t s which occur  female gland  (a).  Intact  X.  D e t a i l of f i g u r e aging  -  (Females)  10 w e e k s . Fig.  of Group I i  the  Compare  X. o f t h e 10 week estradiol  intact  female,  a f t e r prolonged  treatment,  showing  t h e d i r e c t e f f e c t s of t h e hormone  -  t h e m e t a p l a s i a o f t h e squamous e p i t h e l i u m o f the ducts cells  and t h e d e s q u a m a t i o n  i n t o the d u c t s .  41  X.  of e p i t h e l i a l  -94-  PLATE 13 - The E x p e r i m e n t a l G l a n d s Mice Fig.  Fig.  45.  46.  of Group II  The p r e p u t i a l g l a n d  o f t h e 3 week male mouse  w h i c h has  not y e t a t t a i n e d the a d u l t  pattern.  41  o f t h e 6 week  The p r e p u t i a l g l a n d  as m a n i f e s t by h y p e r p l a s i a  Fig.  48.  46.  41  after is  activity  of the a c i n a r  cells  Compare  with  X. o f t h e 6 week male a f t e r  e s t r a d i o l treatment,  i n h i b i t i o n of i t s  showing  n o r m a l g r o w t h by t h e  Note the s m a l l - s i z e stroma,  X.  There  secretory  of the a c i n i .  The p r e p u t i a l g l a n d prolonged  41  treatment.  some i n d i c a t i o n of i n c r e a s e d  figure  45.  structural  o f t h e 6 week male  testosterone  and by h y p e r t r o p h y  normal  the a d u l t  Compare w i t h f i g u r e  prolonged  structural  X.  The p r e p u t i a l g l a n d  pattern. 47.  Weanling  (Males)  male mouse w h i c h e x h i b i t s  Fig.  -  acini,  the u n d e r d e v e l o p e d  Compare w i t h f i g u r e  46.  41  t h e more duct X.  the hormone.  abundant  system.  -95-  PLATE  14 T The E x p e r i m e n t a l G l a n d s Mice  Fig.  49.  50.  The p r e p u t i a l g l a n d  The  41  51.  41 X ,  o f t h e f e m a l e mouse  at  of  t h e f e m a l e mouse  at  o f t h e f e m a l e mouse  at  .  The p r e p u t i a l g l a n d 6 weeks  a f t e r prolonged  u l a t i o n which is the a c i n a r  testosterone  52.  gtim-n  m a n i f e s t by h y p e r p l a s i a  c e l l s , hypertrophy  and by i n c r e a s e d d u c t u l a r  Fig.  Weanling  X.  p r e p u t i a l gland  6 weeks. Fig.  -  (Females)  3 weeks. Fig.  of Group II  of t h e  acini  transformation.  Compare w i t h f i g u r e  50.  The p r e p u t i a l g l a n d  o f t h e f e m a l e mouse  6 weeks,  a f t e r prolonged  Compare w i t h f i g u r e amount  of parenchyma  the very  abundant  the d u c t u l a r  41  X.  estradiol  composed the  of  of  epithelial cells  41  X.  small  small a c i n i ,  metaplasia  e p i t h e l i u m , and t h e  at  treatment.  50 and n o t e t h e  stroma,  of  of  desquamation  i n t o the c e n t r a l  duct.  -96-  PLATE  15 - The E x p e r i m e n t a l G l a n d s  of Group I I I  -  Newborn  Mice Fig.  53.  The p r e p u t i a l g l a n d 3 weeks,  a f t e r prolonged  ulation,  m a n i f e s t by h y p e r p l a s i a  acinar  male g l a n d 54.  in figure  showing  o f t h e hormone.  56.  stim-  the trans-  41  X.  o f t h e - f e m a l e mouse a t testosterone  the s t i m u l a t o r y  Compare w i t h t h e  normal  in figure  41  X.  The p r e p u t i a l g l a n d  o f t h e male mouse  at  a f t e r prolonged  49.  effects  3 week f e m a l e g l a n d  3 weeks,  Fig.  45.  a f t e r prolonged  treatment,  55.  of  at  Compare w i t h t h e n o r m a l 3 week  The p r e p u t i a l g l a n d 3 weeks,  Fig.  testosterone  c e l l s and i n c r e a s e d d u c t u l a r  formation.  Fig.  o f t h e male mouse  estradiol  Compare w i t h f i g u r e  45.  The p r e p u t i a l g l a n d  o f t h e f e m a l e mouse  a t 3 weeks ment.  41  treatment,  a f t e r prolonged  Compare w i t h f i g u r e  X.  estradiol 49.  41  X.  treat-  -97-  PLATE 16 - The Experimental  Glands of Group I I I - Newborn  Mice Fig.  57.  D e t a i l of the normal 3 week p r e p u t i a l (male).  Fig.  58.  122 X.  D e t a i l of f i g u r e  53, the 3 week testosterone-  treated preputial  Fig.  59-  gland  gland  (male).  hypertrophy  of the a c i n i .  figure  122 X.  57.  D e t a i l of f i g u r e treated preputial i t s resemblance,  Note the  Compare with  56, the 3 week e s t r a d i o l gland (female).  Note  g e n e r a l l y , with the normal  3 week gland i n f i g u r e  57.  122 X.  -98-  PLATE 17 Fig.  Histochemistry  60.  Sudan b l a c k B s t a i n i n g ,  demonstrating  d e p o s i t i o n of l i p i d d r o p l e t s cells  o f t h e mouse p r e p u t i a l g l a n d  female).  The l a r g e s t  the d e g e n e r a t i n g acini Fig.  61.  (a).  62.  304  droplets  c e l l s of the  Sudan b l a c k B m e t h o d .  droplets  are merely h e a v i l y 304  The  lipid  o u t l i n e d by  the  X.  throughout  the a c i n a r  t h e mouse p r e p u t i a l g l a n d Again the l a r g e s t degenerating (a).  in  hypertrophied  of  Beaver  showing the d e p o s i t i o n of o i l  0 droplets  acini  (adult  coalesce  D i f f e r e n t i a l s t a i n i n g technique (i960),  acinar  X.  McManus'  stain. Fig.  i n the  the  droplets  c e l l s of t h e 304  X.  (adult  red  cells  of  female).  coalesce  in  hypertrophied  the  PLATE 18 - H i s t o c h e m i s t r y  Pig.  63.  Biebrich at  Scarlet  Fig,  64.  granules  the r a t p r e p u t i a l  in  the a c i p a r c e l l s  gland  65.  (female).  Heidenhain's  granules putial arq  66.  cells  Heidenhain's to  with  304 X.  (female).  method,  Note t h a t t h e g r a n u l e s  approaching  gland  304 X.  method a p p l i e d (male).  of g r a n u l e s .  304 X.  i n the  degeneration-  iron hematoxylin  66.  of the r a t p r e -  and l a r g e i n s i z e  complete a b s e n c e figure  granules  of the r a t p r e p u t i a l  t h e mouse p r e p u t i a l  the  304 X.  hematoxylin  the p r o t e i n  i n the a c i n a r c e l l s  gland  cells  the p e r i n u c l e a r , proteinaceous  most numerous  acinar  acid  i r o n hematoxylin  demonstrating  Fig.  i n the a c i n a r  M a l l o r y ' s phosphot-ungstic  Lillie,  the p e r i n u c l e a r  gland-(female).  method, d e m o n s t r a t i n g  Fig.  o f S p i c e r and  pH 4.95, d e m o n s t r a t i n g  proteinaceous of  method  Note  Compare  -100-  PLATE  19 -  Fig.  Histochemistry  67.  Differential staining (i960)  showing  droplets  (red)  technique  of  Beaver  t h e d e p o s i t i o n o f o i l :red 0 throughout  the a c i n a r  o f t h e mouse p r e p u t i a l g l a n d  cells  (adult,  female).  80 X .  Fig.  68.  D i f f e r e n t i a l s t a i n i n g technique (i960), oil  showing  red 0 droplets  (red)  and dense  which are m a i n l y p e r i n u c l e a r  position  ( b l a c k arrow)  (a).  coalesce  cells  (female).  i n the ducts  The  of the  gland  67 i n w h i c h t h e  are absent.  O i l red 0 s t a i n i n g  in  i n the a c i n a r  Compare w i t h f i g u r e  p r o t e i n granules  of  protein  granules  granules  69.  Beaver  t h e randprn d e p o s i t i o n  of the r a t p r e p u t i a l g l a n d  Fig.  of  f o l l o w e d by  108 X .  .  Heidenhain's  i r o n h e m a t o x y l i n method, d e m o n s t r a t i n g  the  d i s t r i b u t i o n o f dense  (blue-  black)  p r o t e i n granules  and o i l r e d 0 d r o p l e t s  simultaneously  i n the a c i n i of the r a t p r e p u t i a l (male).  136  X.  gland  -101-  A CKN 0WLE D GEMEN T S  Sincere  thanks  a r e e x t e n d e d t o D r . J . M.  f o r h e r a d v i c e and a s s i s t a n c e of t h e s t u d y .  I  i n the g e n e r a l  Taylor  supervision  a l s o w i s h t o t h a n k D r . P. F o r d  and  D r . A . M. P e r k s who p r o v i d e d h e l p f u l c r i t i c i s m and advice.  I  am g r a t e f u l t o D o r o t h y K i e s e r f o r h e r  l a t i o n o f s e v e r a l German a r t i c l e s , t o Daphne  trans-  Hards  for  her v a l u a b l e t e c h n i c a l s k i l l w i t h the h i s t o c h e m i c a l procedures, Finally,  and t o a l l t h e o t h e r p e o p l e who h e l p e d me.  I wish to e s p e c i a l l y thank Denis  encouragement study.  and h e l p t h r o u g h o u t  for  the course  his of  the  -102-  LITERATURE CITED  A r a i , Y . 1 9 6 8 . M e t a p l a s i a i n male r a t r e p r o d u c t i v e a c c e s s o r y g l a n d s i n d u c e d by n e o n a t a l e s t r o g e n t r e a t m e n t . E x p e r e n t i a 24: 180-18,1. B a r n e s , H . H . F . and G . I . M . S w y e r . 1 9 5 1 . E f f e c t of g r o w t h hormone and o f c h o r i o n i c g o n a d o t r o p h i n on p r e p u t i a l g l a n d s of t h e f e m a l e r a t . B r i t . Med. J . 2: 2 0 7 - 2 0 8 , B e a v e r , D.L. 1 9 5 9 . Der E i n f l u s s v e r s c h i e d e n e r P i x i e r u n g s - ^ m i t t e l a u f das S t r t i k t u r b i l d d e r P r a p u t i a l d r u s e n d e r Ratte. Z t s c h r . Z e l l f o r s c h . m i k r . Anat.- 5 1 ; 88-96 B e a v e r , D.L. 1 9 6 0 . The r e - e v a l u a t i o n of t h e r a t p r e p u t i a l g l a n d as a " d i c r i n e " o r g a n f r o m t h e s t a n d p o i n t of i t s m o r p h o l o g y , h i s t o c h e m i s t r y , and p h y s i o l o g y . J . Exp. Z o o l . 143: 15,3-173. Bronson, F.H. preputial  1966. A s e x a t t r a c t a n t f u n c t i o n f o r mouse glands. Amer. Z o o l . 6: 535. Abstract.  B r o n s o n , F . H . , C - P . Dagg and G.D. S n e l l . 1 9 6 6 . Reproduction. I n B i o l o g y of the L a b o r a t o r y Mouse, (E. G r e e n , e d . ) , 2nd e d i t i o n , The J a c k s o n L a b o r a t o r y . B r o o k s , S . C . , J . J . L a l i c h , C.A. Baumann. 1 9 5 6 . Skin sterolsXI A d i r e c t d e m o n s t r a t i o n o f f a s t - a c t i n g s t e r o l s i n t h e sebaceous g l a n d s . Am. J . P a t h o l . 3 2 : 1205-1213. B u r d i c k , H.O. and E. Gamon. 1 9 4 1 , Response of the p r e p u t i a l g l a n d s of t h e f e m a l e mouse t o t e s t o s t e r o n e p r o p i o n a t e . E n d o c r i n o l o g y 28: 677-679. B u r n s , R.K. 1 9 6 1 . R o l e of hormones i n t h e d i f f e r e n t i a t i o n of s e x . I n Sex and I n t e r n a l S e c r e t i o n s , " V o l . 1, ( V . C . Y o u n g , e d . ) , 3 r d e d i t i o n . The W i l l i a m s and ' W i l k i n s Co. p p . 7 6 - 1 5 8 . B u r r o w s , H. 1935. P a t h o l o g i c a l c o n d i t i o n s induced by c e s t r o g e n i c compounds i n t h e c o a g u l a t i n g g l a n d and prostate of t h e mouse. Am. J . C a n c e r 2 3 ( 3 ) : 4 9 0 - 5 1 2 . . B u r r o w s , H. 1937. Changes i n d u c e d b y o e s t r o n e i p t h e b u l b o - u r e t h r a l ( C p w p e r ' s ) g l a n d s o f t h e male mouse. J , Path. & Bact. 44(2): 481-483.  -103-  B u s c h k e , V. 1933. Die Hautdrusenorgan (Hardersche Drflsen, Inguinaldrusen, Pr&pufialdrusen, Analdrusen, Kaudaldrtisen, K i e f e r d r i l s e n ) d e r ' L a b o r a t o r i u m s n a g e t i e r e und d i e F r a g e i h r e r A b h & n g i g k e i t v o n den G e s e h l e c h t s d r t l s e n . Ztschr. Zellforsch. m i k r . A n a t . 18: 217-243. C h a y e n , J . , L. B i t e n s k y , R.G. B u t c h e r , L.W. P o u l t e r . A Guide to P r a c t i c a l H i s t o c h e m i s t r y . University Aberdeen. C o c k r u m , E..L. 1 9 6 2 . P r e s s C o . , N.Y.  Introduction  t o Mammalogy.  1969. Press,  Ronald  C u l l i n g , C . F . A . 1957. Handbook o f H i s t o p a t h o l o g i c a l T e c h n i q u e , 2nd- e d i t i o n . B u t t e r w o r t h & Co. L t d . , L o n d o n . E b l i n g , F . J . 1948. Sebaceous g l a n d s . I. The e f f e c t o f s e x hormones on t h e s e b a c e o u s g l a n d s o f t h e f e m a l e a l b i n o rat. J . E n d o c r i n . 5: 297-302. E n g l e , E . T . and J . R o s a s c o . 1927. The age o f t h e a l b i n o mouse .at n o r m a l s e x u a l m a t u r i t y . A n a t . Rec. 36: 383-388. F r e e m a n , J . J . , R. H i l f , A . J , I o v i n o , I. M i c h e l . 1964. E f f e c t s p f s t e r o i d s upon t h e w e i g h t , p r o t e i n and n u c l e i c a c i d c o n c e n t r a t i o n of t h e p r e p u t i a l g l a n d s the female r a t . Endocrin. 74(6): 990-993. G a l l o , D.G. gland.  of  1966. L i p i d s y n t h e s i s by the r a t p r e p u t i a l Fed. Proc. 2 5 ( 2 , i ) : 766.  G a u n t , S . L . 1967. C l a s s i f i c a t i o n and e f f e c t o f t h e p r e p u t i a l pheromone i n t h e mouse. Amer. Z o o l . 7: Abstract. G r e e p , R.O. 1966. Book Co.  ed. Histology,  713  2nd e d i t i o n . M c G r a w - H i l l  H a r d y , M.H. 1949. The d e v e l o p m e n t o f mouse h a i r i n v i t r o w i t h some o b s e r v a t i o n s on p i g m e n t a t i o n . J . Anat. London 83: 364-384. i  H a s k i n , D., N. L a s h e r , S. Rothman. 1 9 5 3 . Some e f f e c t s of ACTH, c o r t i s o n e , p r o g e s t e r o n e , and t e s t o s t e r o n e on sebaceous g l a n d s i n the w h i t e r a t . J . I n v e s t . Dermat. 20: 207-212.  -104-  H u g g i n s , C , F. P a r s o n s , E. J e n s e n . 1 9 5 5 . P r o m o t i o n of g r o w t h of p r e p u t i a l g l a n d s by s t e r o i d s and t h e p i t u i t a r y g r o w t h hormone. E n d o c r i n . 57: 25-32. K a n d u t s c h , A . A . . 1964. S t e r o l m e t a b o l i s m i n s k i n and epidermis. I n The E p i d e r m i s , (V. M o n t a g n a and V . C . L o b i t z , e d s . ) A c a d e m i c P r e s s , N.Y. p p . 4 9 3 - 5 1 0 . K o r e n c h e v s k y , V. and M. D e n n i s o n . 1 9 3 4 a . The e f f e c t o f o e s t r o n e on n o r m a l and c a s t r a t e d r a t s . Biochem. J . 1474-1485.  28:  K o r e n c h e v s k y , V. and M. D e n n i s o n . 1 9 3 4 b . The e f f e c t on . male r a t s of t h e s i m u l t a n e o u s a d m i n i s t r a t i o n o f male and f e m a l e s e x u a l hormones and t h e r e l a t i o n t o t h e a s s a y of the hormones. Biochem. J . 28: 1486-1499. K o r e n c h e v s k y , V. and M. D e n n i s o n . 1 9 3 5 . Histological changes i n the organs of r a t s i n j e c t e d w i t h o e s t r o n e a l o n e o r s i m u l t a n e o u s l y w i t h o e s t r o n e and t e s t i c u l a r hormone. J . P a t h . & B a c t . 4 1 : 323-337. K o r e n c h e v s k y , V. and M. D e n n i s o n . 1 9 3 6 a . The h i s t o l o g y of the sex organs of o v a r i e c t o m i z e d r a t s t r e a t e d w i t h male o r f e m a l e s e x hormone a l o n e o r w i t h b o t h simultaneously. J , P a t h . & Bact. 42: 91-104. K o r e n c h e v s k y , V. and M. D e n n i s o n . 1936b. The h i s t o l o g i c a l c h a n g e s i n t h e sex o r g a n s o f s p a y e d r a t s i n d u c e d b y t e s t o s t e r o n e and o e s t r o n e . J . Path. & Bact. 43: 345-356. L i l l i e , R.D. 1965. Histochemistry,  H i s t o p a t h o l o g i c a l T e c h n i c and P r a c t i c a l 3 r d e d i t i o n . M c G r a w - H i l l Book Co.  L y n e , A . G . 1966. The d e v e l o p m e n t J . S c i . 2 8 ( 1 0 ) : ,370-377.  of h a i r f o l l i c l e s .  Aust.  M i r s k a i a , L. and F . A . E . Crew. 1930. On t h e g e n e t i c n a t u r e of t h e t i m e of a t t a i n m e n t o f p u b e r t y i n t h e f e m a l e mouse. Q u a r t . J . E x p . P h y s i o l . 20: 299-304. M o n t a g n a , ¥ . and H. C h a s e . 1950. R e d i f f e r e n t i a t i o n of s e b a c e o u s g l a n d s i n t h e mouse a f t e r t o t a l e x t i r p a t i o n with methylcholanthrene. A n a f . R e c . 107: 83-91. M o n t a g n a , ¥ . and C R . N o b a c k . 1946. The h i s t o l o g y p f t h e p r e p u t i a l g l a n d o f the. r a t . A n a t . R e c . 96: 41-54.  -105-  M o o r e , C R . and D. P r i c e . 1 9 3 2 . Gonad hormone f u n c t i o n s and t h e r e c i p r o c a l i n f l u e n c e b e t w e e n gonads and h y p o p h y s i s w i t h i t s b e a r i n g on t h e p r o b l e m o f s e x hormone a n t a g o n i s m . Am. J . A n a t . 50: 13-71. M o o r e , C R . and D. P r i c e . 1938. Some e f f e c t s o f t e s t o s t e r o n e and t e s t o s t e r o n e p r o p i o n a t e i n t h e r a t . Anat. Rec. 71: 59-78. M o o r e , R.A. 1936. The e v o l u t i o n and i n v o l u t i o n o f prostate gland. Am. J . P a t h . 1 2 : 599-624.  the  N o b l e , R.L. and J . B . C o l l i p . 1 9 4 1 A possible direct c o n t r o l o f t h e p r e p u t i a l g l a n d s of t h e f e m a l e r a t b y t h e p i t u i t a r y g l a n d and i n d i r e c t e f f e c t s p r o d u c e d t h r o u g h t h e a d r e n a l s and gonads b y augmented p i t u i t a r y extracts. E n d o c r i n . 29: 943-951. O d l a n d , G . P . 1966. Skin. I n H i s t o l o g y , (R.O. G r e e p , 2nd e d i t i o n . M c G r a w - H i l l Book Co. p p . 4 2 0 - 4 4 5 . P a r k e s , A.S.. 1 9 5 0 . Recent Progress  ed.),  A n d r o g e n i c a c t i v i t y of the o v a r y . i n Hormone R e s e a r c h 5: 101-114.  P a t t e r s o n , J . F , I960. B i o s y n t h e s i s of squalene by preputial gland. P r o c . S o c . E x p t l . B i o l . Med. 461-463.  rat 105:  P e a r s e , A . G . E . 1961. H i s t o c h e m i s t r y . T h e o r e t i c a l and A p p l i e d , 2nd e d i t i o n . L i t t l e , Brown & Co. P e a r s e , A . G . E . 1968. Histochemistry. T h e o r e t i c a l and A p p l i e d , V o l . 1, 3 r d e d i t i o n . L i t t l e , Brown & Co. P r i c e , D. 1936. N o r m a l d e v e l o p m e n t o f t h e p r o s t a t e and s e m i n a l v e s i c l e s of the r a t w i t h a s t u d y of e x p e r i m e n t a l p o s t - n a t a l m o d i f i c a t i o n s . Am. J . A n a t . 60: 79-127. P r i c e , D. and E. O r t i z . 1944 The r e l a t i o n o f age t o r e a c t i v i t y i n the r e p r o d u c t i v e system of the r a t . E n d o c r i n . 34: 215-239. P r i c e , D. and H.G. W i l l i a m s - A s h m a n . 1 9 6 1 . The a c c e s s o r y r e p r o d u c t i v e g l a n d s o f mammals. I n Sex and I n t e r n a l S e c r e t i o n s , V o l . 1, ( W . C Y o u n g , e d . ) , 3 r d e d i t i o n . The W i l l i a m s and W i l k i n s Co. p p . 3 6 6 - 4 4 8 . R e n n e l s , E . G . , M. H e s s , J . C . F i n e r t y . 1 9 5 3 . Response of p r e p u t i a l and a d r e n a l g l a n d s o f t h e r a t t o sex h o r m o n e s . P r o c . Soc. E x p t l . B i o l . (N.Y.) 82: 304-309.  -106-  S a l m o n , U . J . 1938. The e f f e c t o f t e s t o s t e r o n e p r o p i o n a t e on t h e g e n i t a l t r a c t o f t h e immature f e m a l e r a t . E n d o c r i n . 23: 779-783. S c h a f f e r , J . 1933. D i e V o r h a u t d r t i s e n v o n Maus und R a t t e . Z t s c h r . m i k r . A n a t . F o r s c h . 34: 1-22. S n e l l , G.D. 1 9 4 1 . Reproduction. In B i o l o g y of the L a b o r a t o r y M o u s e , (G.D. S n e l l , e d . ) , B l a k i s t o n , P h i l a d e l p h i a , pp. 55-88. Tepperman, J . 1968. M e t a b o l i c and E n d o c r i n e P h y s i o l o g y , 2nd e d i t i o n . Y e a r Book M e d i c a l P u b l i s h e r s , I n c . Thung, P . J . , L.M. B o o t , 0 . M i i h l b o c k . 1956. S e n i l e changes i n t h e e s t r u s c y c l e and i n o v a r i a n s t r u c t u r e i n some i n b r e d s t r a i n s of m i c e . A c t a E n d o c r i n o l o g i c a 23: 8-32. V o s s , H.E. 1 9 3 2 . D i e P r f t p u t i a l d r t l s e n d e r Maus i n i h r e r A b h f t n g i g k e i t v o n hormon des H o d e n s . Zeitschr. Zellforsch. m i k r . A n a t . 14: 200-221.  

Cite

Citation Scheme:

        

Citations by CSL (citeproc-js)

Usage Statistics

Share

Embed

Customize your widget with the following options, then copy and paste the code below into the HTML of your page to embed this item in your website.
                        
                            <div id="ubcOpenCollectionsWidgetDisplay">
                            <script id="ubcOpenCollectionsWidget"
                            src="{[{embed.src}]}"
                            data-item="{[{embed.item}]}"
                            data-collection="{[{embed.collection}]}"
                            data-metadata="{[{embed.showMetadata}]}"
                            data-width="{[{embed.width}]}"
                            async >
                            </script>
                            </div>
                        
                    
IIIF logo Our image viewer uses the IIIF 2.0 standard. To load this item in other compatible viewers, use this url:
http://iiif.library.ubc.ca/presentation/dsp.831.1-0101849/manifest

Comment

Related Items