Horng Jiun-Lin, Chao Pei-Lin, Chen Po-Yen, Shih Tin-Han, Lin Li-Yih
Department of Anatomy and Cell Biology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan.
Department of Life Science, National Taiwan Normal University, Taipei, Taiwan.
PLoS One. 2015 Aug 19;10(8):e0136440. doi: 10.1371/journal.pone.0136440. eCollection 2015.
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 ion-selective 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.
哺乳动物水通道蛋白1(AQP1)作为一种负责水和二氧化碳运输的膜通道已广为人知。斑马鱼AQP1a.1(哺乳动物AQP1的同源物)最近在胚胎的离子细胞中被发现;然而,其在离子细胞中的作用仍不清楚。在本研究中,我们假设斑马鱼AQP1a.1通过促进水和二氧化碳的扩散参与离子细胞的酸分泌。实时定量PCR结果显示,暴露于1%二氧化碳高碳酸血症3天可诱导胚胎中AQP1a.1的mRNA水平升高。原位杂交和免疫组化结果表明,AQP1a.1转录本在分泌酸的离子细胞,即富含H⁺-ATP酶(HR)的细胞中高表达。应用扫描离子选择性电极技术(SIET)分析胚胎中单个离子细胞的二氧化碳诱导的H⁺分泌。短暂加载二氧化碳(1%,10分钟)后,HR细胞的H⁺分泌显著增加。用吗啉代寡核苷酸敲低AQP1a.1可使HR细胞的H⁺分泌减少约一半,并限制了二氧化碳刺激的增加。此外,暴露于AQP抑制剂(对氯汞苯甲酸)10分钟也抑制了二氧化碳诱导的H⁺分泌。本研究结果支持了我们的假设,并为AQP1在二氧化碳运输中的生理作用提供了体内证据。