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1 Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan; Institute of Fisheries Science, National Taiwan University, Taipei, Taiwan
2 Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan
3 Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan; Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan
* To whom correspondence should be addressed. E-mail: pphwang{at}gate.sinica.edu.tw.
The purpose of the present work was to study the possible role of the ECaC (epithelial calcium channel) in the Ca2+ uptake mechanism in developing zebrafish (Danio rerio). Using rapid amplification of cDNA ends (RACE), full-length cDNA encoding the ECaC of zebrafish was cloned and sequenced. The cloned zECaC was 2578 base pairs in length, and encoded a protein of 709 amino acids that showed up to 73% identity with previously described vertebrate ECaCs. The zECaC was found to be expressed in all tissues examined, and began to be expressed in the skin covering the yolk sac of embryos at 24 h post-fertilization (hpf). zECaC-expressing cells expanded to cover the skin of the entire yolk sac following embryonic development, and began to occur in the gill filaments at 96 hpf, and thereafter zECaC-expressing cells rapidly increased in both gills and yolk sac skin. In correspondence to the profile of ECaC expression, the Ca2+ influx and content began to increase at 36~72 hpf. Incubating zebrafish embryos in low-Ca2+ (0.02 mM) fresh water caused upregulation of the whole-body Ca2+ influx and zECaC expression in both gills and skin. Co-localization of zECaC mRNA and the Na+/K+-ATPase 
subunit (a marker for mitochondria-rich, MR, cells) indicated that only a portion of the MR cells expressed zECaC mRNA. These results suggest that the zECaC plays a key role in Ca2+ absorption in developing zebrafish.
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