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1 Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan - Republic of China; Institute of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung, Taiwan - Republic of China
2 Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan - Republic of China; Institute of Fishery Science, National Taiwan University, Taipei, Taiwan - Republic of China
3 Department of Aquatic Bioscience, University of Tokyo, Tokyo, Bunkyo, Japan
4 Institute of Zoology, Academia Sinica, Taipei, United States; Institute of Zoology, Academia Sinica, Nankang, Taipei, 11529
* To whom correspondence should be addressed. E-mail: pphwang{at}gate.sinica.edu.tw.
In mammalian nephrons, most of the Na+ and HCO3- is reabsorbed by proximal tubular cells in which the Na+/H+ exchanger 3 (NHE3) is the major player. The roles of NHEs in Na+ uptake/acid-base regulation in freshwater (FW) fish gills are still being debated. In the present study, functional genomic approaches were used to clone and sequence the full-length cDNAs of the nhe family from zebrafish (Danio rerio). A phylogenetic tree analysis of the deduced amino acid sequences showed that zNHE1~8 are homologous to their mammalian counterparts. By RT-PCR analysis and double/triple in situ hybridization/immunocytochemistry, only zNHE3b was expressed in zebrafish gills, and was colocalized with V-H+-ATPase but not with Na+-K+-ATPase, indicating that HR (H+-ATPase-rich) cells specifically express NHE3b. A subsequent quantitative RT-PCR analysis demonstrated that acclimation to low-Na+ FW caused up-regulation and down-regulation of the expressions of znhe3b and zatp6v0c (H+-ATPase C-subunit), respectively, in gill HR cells, while acclimation to acidic FW showed reversed effects on the expressions of these two genes. In conclusion, both NHE3b and H+-ATPase are probably involved in Na+ uptake/acid-base regulation in zebrafish gills, like mammalian kidneys, but the partitioning of these two transporters may be differentially regulated depending on the environmental situation in which fish are acclimatized.
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