UBC Theses and Dissertations
Properties of ion and fluid transport and control in hindgut of the desert locust (Schistocerca gregaria) Lechleitner, Richard August
Previous studies of selective reabsorption in insect excretory system have concentrated almost exclusively on the rectum, while the role of the ileum has been assumed to be minor. The properties and control of solute and fluid transport in two segments of the hindgut, the ileum and rectum, from the desert locust (Schistocerca gregaria) have been studied and compared in vitro using everted sac and flat sheet preparations. Everted sacs of locust ileum transported fluid from the lumen side to hemocoel side over a 5 h period at near constant rates of 3.0 to 3.5 μL-h⁻¹-ileum⁻¹ and tissue volume did not change. Inhibition by azide indicated metabolic dependence of fluid transport. Fluid absorption occurred against osmotic concentration differences of up to 600 mosmol. Fluid transport was stimulated by cAMP, both nervous and glandular lobes of corpus cardiacum (CC), and fifth ventral ganglia (VG) in a dose-dependent manner. All stimulants caused ilea to absorb against larger osmotic concentration differences than unstimulated sacs. The ileal absorbate remained hyperosmotic to the luminal saline under all conditions and stimulants increased absorbate osmolality. Unstimulated fluid transport was supported at 50% of control levels by any one of Na+, K+, or CI⁻. Stimulation of fluid transport by CC or VG was dependent on CI⁻ and maximal stimulation occurred when the Na+:K+ ratio was 1:1. Cyclic AMP, CC and VG all stimulated Na+,K+ and CI⁻ absorption across everted ileal sacs. This is the first direct demonstration that Na+ reabsorption is controlled in insect excretory systems. Stimulation resulted in a decrease in absorbate HCO₃⁻ levels and pH concurrently with an increase in absorbate CI⁻ levels. Stimulation of fluid transport was associated with a 3-fold increase in transepithelial potential (hemocoel negative) suggesting stimulation of electrogenic anion (CI⁻) movement to the hemocoel. Net Na+ absorption occurs largely by electroneutral active transport. NH₄+/Na+ exchange may account for one-third of stimulated net Na+ flux. Extracts from both CC and VG stimulated fluid, K+, and CI⁻ transport across everted rectal sacs, but only a small stimulation of Na+ flux was observed which was an order of magnitude less than that observed for stimulated ilea (0.4 versus 5.1 μequiv-h⁻¹ -cm⁻²). Unlike the rectum, the ileum did not transport proline transepithelially and ileal fluid transport was not stimulated by increasing concentrations of proline in the bathing saline. Rectal fluid transport was stimulated 50% by increasing external proline concentration from 1 to 80 mM. Stimulation of rectal fluid transport by proline also occurred in the absence of Na+, K+, and CI⁻ and occurred against larger osmotic concentration differences. These results are consistent with previous reports of a high capacity transport system for proline in locust rectum. The presence of anion-stimulated ATPase and Na+,K+-ATPase in locust hindgut was also investigated. Anion-stimulated ATPase activities were observed in microsomal fractions of both rectum and ileum. Microsomal fractions from both tissues had enriched specific activities of several plasma membrane marker enzymes and decreased activities of two mitochondrial markers as compared to homogenate enzyme activities. Na+,K+-ATPase activity was 20-fold higher in the rectum than in the ileum, associated with the greater development of the basolateral membrane in the rectum. Overall the results suggest that ion and fluid reabsorption in the locust ileum is much more important in the excretory process than previously supposed. Moreover, this reabsorption was shown to be under neuroendocrine control.
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