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Expression and function of ryanodine receptors in human melanocytes
Author(s) -
Kang HeeYoung,
Kim NamSoo,
Lee ChinOk,
Lee JiYeoun,
Kang Won Hyoung
Publication year - 2000
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/1097-4652(200011)185:2<200::aid-jcp4>3.0.co;2-6
Subject(s) - ryanodine receptor , ryr1 , gene isoform , immunocytochemistry , biology , cytoplasm , intracellular , microbiology and biotechnology , receptor , melanocyte , biochemistry , endocrinology , gene , genetics , melanoma
We investigated the expression of ryanodine receptors (RyRs) in cultured human melanocytes with immunocytochemistry and reverse transcriptase–polymerase chain reaction. With the use of a monoclonal antibody, RyR immunoreactivity was detected in the cytoplasm of melanocytes, and was further confirmed by RT‐PCR assay. The PCR products were cut with restriction enzymes specific for each RyR isoform. Using the RyR1‐specific restriction enzyme Sac I yielded fragments of 300, 100, and 130 base pairs, consistent with the expression of RyR1 isoforms. The function of RyR in Ca 2+ signaling was investigated using single‐cell fura‐2 imaging. Ryanodine (1 to ∼100 μM) induced significant elevation of cytoplasmic Ca 2+ in single human melanocytes in a dose‐dependent manner. The ryanodine‐induced [Ca 2+ ] i increase was inhibited by neomycin. Furthermore, ryanodine inhibited proliferation and stimulated pigmentation of human melanocytes. This study demonstrates that the RyR1 isoform is expressed in cultured human melanocytes, and suggests that the RyR may be involved in regulating the intracellular Ca 2+ responses involved in proliferation and pigmentation of cultured human melanocytes. J. Cell. Physiol. 185:200–206, 2000. © 2000 Wiley‐Liss, Inc.