Abstract
The newly identified metazoan Na+/H+ antiporter (NHA) family is represented by two paralogues, AgNHA1 and AgNHA2, in the genome of the African malaria mosquito, Anopheles gambiae. Both antiporters are postulated to be electrophoretic i.e. voltage-driven. AgNHA1 was first cloned from An. gambiae larvae and immunolocalized with respect to the H+ V-ATPase by the Harvey laboratory. Little is known about the properties of NHA1s; attempts to characterize AgNHA1 in Na+/H+ exchanger (NHE)-lacking Chinese hamster ovary cells and in yeast cells or frog oocytes were unsuccessful. Even less is known about AgNHA2. It is predicted to have a relative molecular mass of ∼60kDa and shares 30.5% amino acid identity with AgNHA1. Immunolocalization images show AgNHA2 on the apical plasma membrane of stellate cells in Malpighian tubules of An. gambiae larvae and adults. When heterologously expressed in a mutant strain of the yeast, Saccharomyces cerevisiae, which lacks endogenous cation/proton antiporters and pumps, AgNHA2 enhanced repression of growth by the alkali metal cations, Li+, Na+, or K+ and enhanced Li+ accumulation. The yeast growth studies invite the speculation that AgNHA2 is an electrophoretic antiporter with a stoichiometry of nNa+ to 1H+ with n > 1. Immunolocalization images provide direct evidence that H+ V-ATPase is co-localized with AgNHA1 on the apical membrane of principal cells but it is not present in the stellate cells where AgNHA2 is localized apically. These results are consistent with the notion that the outside positive voltage that the H+ V-ATPase generates across the apical membrane of principal cells appears with but little attenuation across the apical membrane of stellate cells. This immunolocalization pattern is consistent with the hypothesis that the voltage acts via AgNHA1 to drive nH+ into the principal cells and Na+ out to the lumen and acts via AgNHA2 to drive nNa+ into the stellate cells and H+ out to the lumen. Precious Na+ is then retained by ejection into the blood via a basal Na+/K+-ATPase. Localizations of anion transporters and their functions in stellate and principal cells are described by Linser, Romero and associates in this volume. The role that the electrogenic H+ V-ATPase and the electrophoretic cationic and anionic transporters play in ion homeostasis is incorporated into a model for Malpighian tubule cells of larval mosquitoes.
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