UBC Theses and Dissertations

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UBC Theses and Dissertations

Intercellular junctions and tubulobulbar complexes between sertoli cells in primary culture and in vivo Sriram, Aarati


Tubulobulbar complexes (TBCs) are actin-rich structures that form at intercellular junctions in the seminiferous epithelium of the mammalian testis. Massive intercellular junctions form at the base of the epithelium where Sertoli cells are connected to each other, and at the apex where Sertoli cells are connected to mature spermatids. TBCs are proposed to be responsible for internalizing these intact junctions during spermiation at the apex and during the translocation of spermatocytes from basal to adluminal compartments near the base of the epithelium. A growing body of evidence indicates that apical TBCs internalize tissue specific adhesion junctions at the apex of the epithelium and are involved in sperm release. In comparison, relatively little is known about basal TBC function and spermatocyte translocation. This thesis explores the hypothesis that the presence of spermatogenic cells influences the structure of TBCs at basal junctions between Sertoli cells in vitro. A primary Sertoli-germ cell co-culture system was optimized and used to explore TBC structure in vitro. In the seminiferous epithelium, if basal TBCs are responsible for internalizing junctions, then interfering with TBC structure or formation should lead to delayed translocation of spermatocytes and increased mass of basal intercellular junctions. An in vivo ribonucleic acid interference (RNAi) procedure was optimized to knockdown cortactin, a component of TBCs, and the morphological differences on basal TBCs were observed. This work is a necessary prelude to future work to evaluate the role of basal TBCs during spermatocyte translocation. Finally, this thesis shows that disruption of actin at basal TBCs results in the same altered TBC structure as has been observed at apical sites.

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