Am J Physiol Cell Physiol AJP: Endocrinology and Metabolism
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Am J Physiol Cell Physiol 294: C1250-C1260, 2008. First published March 5, 2008; doi:10.1152/ajpcell.00021.2008
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MEMBRANE TRANSPORTERS, ION CHANNELS, AND PUMPS

Carbonic anhydrase 2-like a and 15a are involved in acid-base regulation and Na+ uptake in zebrafish H+-ATPase-rich cells

Tzung-Yi Lin,1,2,* Bo-Kai Liao,1,* Jiun-Lin Horng,1,3 Jia-Jiun Yan,1 Chung-Der Hsiao,1 and Pung-Pung Hwang1

1Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan; 2Institute of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung, Taiwan; and 3Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan, Republic of China

Submitted 15 January 2008 ; accepted in final form 4 March 2008

H+-ATPase-rich (HR) cells in zebrafish gills/skin were found to carry out Na+ uptake and acid-base regulation through a mechanism similar to that which occurs in mammalian proximal tubular cells. However, the roles of carbonic anhydrases (CAs) in this mechanism in zebrafish HR cells are still unclear. The present study used a functional genomic approach to identify 20 CA isoforms in zebrafish. By screening with whole mount in situ hybridization, only zca2-like a and zca15a were found to be expressed in specific groups of cells in zebrafish gills/skin, and further analyses by triple in situ hybridization and immunocytochemistry demonstrated specific colocalizations of the two zca isoforms in HR cells. Knockdown of zca2-like a caused no change in and knockdown of zca15a caused an increase in H+ activity at the apical surface of HR cells at 24 h postfertilization (hpf). Later, at 96 hpf, both the zca2-like a and zca15a morphants showed decreased H+ activity and increased Na+ uptake, with concomitant upregulation of znhe3b and downregulation of zatp6v1a (H+-ATPase A-subunit) expressions. Acclimation to both acidic and low-Na+ fresh water caused upregulation of zca15a expression but did not change the zca2-like a mRNA level in zebrafish gills. These results provide molecular physiological evidence to support the roles of these two zCA isoforms in Na+ uptake and acid-base regulation mechanisms in zebrafish HR cells.

ionocytes; Na+/H+ exchanger; skin; gill; embryo



Address for reprint requests and other correspondence: P.-P. Hwang, Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan (e-mail: pphwang{at}gate.sinica.edu.tw)




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