Identification and apical membrane localization of an electrogenic Na+/Ca2+ exchanger NCX2a likely to be involved in renal Ca2+ excretion by seawater fish

摘要

Seawater (SW) contains ∼10 mM Ca²⁺, yet marine fish must drink seawater as their major water source. Thus marine teleosts fish need to excrete Ca²⁺ to maintain whole body Ca²⁺ homeostasis. In the intestine, seawater Ca²⁺ interreacts with epithelial-secreted HCO3⁻ by the intestinal epithelium, and the resulting CaCO3 precipitates, which is rectally excreted. Recently the transporters involved in intestinal HCO3⁻ secretion were identified. Ca²⁺ is also excreted by the kidney, but the protein(s) involved in renal Ca²⁺ excretion have not been identified. Here we identified a candidate transporter by using SW pufferfish torafugu (Takifugu rubripes) and its closely related euryhaline species mefugu (Takifugu obscurus), which are becoming useful animal models for studying molecular mechanisms of seawater adaptation. RT-PCR analyses of Na⁺/Ca²⁺ exchanger (NCX) family members in various torafugu tissues demonstrated that only NCX2a is highly expressed in the kidney. Renal expression of NCX2a was markedly elevated when mefugu were transferred from freshwater to seawater. In situ hybridization and immunohistochemical analyses indicated that NCX2a is expressed in the proximal tubule at the apical membrane. NCX2a, expressed in Xenopus oocytes, conferred [Ca²⁺]out- and Na⁺-dependent currents. These results suggest that NCX2a mediates renal Ca²⁺ secretion at the apical membrane of renal proximal tubules and has an important role in whole body Ca²⁺ homeostasis of marine teleosts.