Mechanisms of transport of H+, Na+ and K+, across the distal gastric caecum of larval Aedes aegypti

Measured changes in ion fluxes, transepithelial potential (TEP) and basolateral membrane potential (V_b) in response to ion transporter inhibitors were used to assess the mechanisms of transport of H⁺, Na⁺ and K⁺, across the distal gastric caecum of larval Aedes aegypti, a vector of yellow fever. Preparations were stimulated with 5-hydroxytryptamine (5-HT, 10^-6 M) in order to maintain stable rates of H⁺, Na⁺, and K⁺ transport across the distal caecum. Transepithelial potential (TEP), basolateral membrane potential (V_b), and H⁺, Na⁺ and K⁺ fluxes all declined after the addition of a vacuolar-type H⁺-ATPase (VA) inhibitor, n-ethlymaleimide (NEM), consistent with a primary role for VA in energizing ion transport across the distal gastric caecum. Amiloride also inhibited H⁺, Na⁺, and K⁺ fluxes, consistent with an apically expressed VA that is coupled to a cation:H⁺ antiporter (AeNHE8), analogous to the coupling of apical VA and cation:nH⁺ antiporter in Malpighian tubules. A working model of transport of H⁺, Na⁺ and K⁺ across the distal gastric caecum proposes that coupling of VA and AeNHE8 in the apical membrane leads to the removal of intracellular Na⁺ or K⁺, thus creating favourable ion gradients to promote the activity of two transporters in the basal membrane, cation:H⁺ antiporter (AeNHE3) and a bumetanide-sensitive cation chloride cotransporter (CCC).