Aquaporin 1 is involved in acid secretion by ionocytes of zebrafish embryos through facilitating CO₂ transport

Mammalian aquaporin 1 (AQP1) is well known to function as a membrane channel for H₂O and CO₂ transport. Zebrafish AQP1a.1 (the homologue of mammalian AQP1) was recently identified in ionocytes of embryos; however its role in ionocytes is still unclear. In this study, we hypothesized that zebrafish AQP1a.1 is involved in the acid secretion by ionocytes through facilitating H₂O and CO₂ diffusion. A real-time PCR showed that mRNA levels of AQP1a.1 in embryos were induced by exposure to 1% CO₂ hypercapnia for 3 days. In situ hybridization and immunohistochemistry showed that the AQP1a.1 transcript was highly expressed by acid-secreting ionocytes, i.e., H⁺-ATPase-rich (HR) cells. A scanning ionselective electrode technique (SIET) was applied to analyze CO₂-induced H⁺ secretion by individual ionocytes in embryos. H⁺ secretion by HR cells remarkably increased after a transient loading of CO₂ (1% for 10 min). AQP1a.1 knockdown with morpholino oligonucleotides decreased the H⁺ secretion of HR cells by about half and limited the CO₂ stimulated increase. In addition, exposure to an AQP inhibitor (PCMB) for 10 min also suppressed CO₂-induced H⁺ secretion. Results from this study support our hypothesis and provide in vivo evidence of the physiological role of AQP1 in CO₂ transport.