Alkalinization in the isolated and perfused anterior midgut of the larval mosquito, Aedes aegypti
Moffett, Stacia B.
Moffett, David F.
Journal of Insect Science
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In the present study, isolated midguts of larval Aedes aegypti L. (Diptera: Culicidae) were mounted on perfusion pipettes and bathed in high buffer mosquito saline. With low buffer perfusion saline, containing m-cresol purple, transepithelial voltage was monitored and luminal alkalinization became visible through color changes of m-cresol purple after perfusion stop. Lumen negative voltage and alkalinization depended on metabolic energy and were stimulated in the presence of serotonin (0.2 μmol l−1). In some experiments a pH microelectrode in the lumen recorded pH values up to 10 within minutes after perfusion stop. The V-ATPase inhibitor concanamycin (50 μmol l−1) on the hemolymph side almost abolished Vte and inhibited luminal alkalinization. The carbonic anhydrase inhibitor, methazolamide (50 μmol l−1), on either the luminal or hemolymph-side, or the inhibitor of anion transport, DIDS (1 mmol l−1) on the luminal side, had no effect on Vte or alkalinization. Cl− substitution in the lumen or on both sides of the tissue affected Vte, but the color change of m-cresol purple was unchanged from control conditions. Hemolymph-side Na+ substitution or addition of the Na+/H+ exchange inhibitor, amiloride (200 μmol l−1), reduced Vte and luminal alkalinization. Luminal amiloride (200 μmol l−1) was without effects on Vte or alkalinization. High K+ (60 mmol l−1) in the lumen reduced Vte without affecting alkalinization. These results indicate that strong luminal alkalinization in isolated and perfused anterior midgut of larval A. aegypti depends on basolateral V-ATPase, but is apparently independent of carbonic anhydrase, apical Cl−/HCO3− exchange or apical K+/2H+ antiport.