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TRPV3 Channel Negatively Regulates Cell Cycle Progression and Safeguards the Pluripotency of Embryonic Stem Cells
Author(s) -
Lo Iek Chi,
Chan Hing Chung,
Qi Zenghua,
Ng Kwun Lam,
So Chun,
Tsang Suk Ying
Publication year - 2016
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/jcp.25086
Subject(s) - microbiology and biotechnology , embryonic stem cell , cell cycle , chemistry , western blot , stem cell , biology , cell , biochemistry , gene
Embryonic stem cells (ESCs) have tremendous potential for research and future therapeutic purposes. However, the calcium handling mechanism in ESCs is not fully elucidated. Aims of this study are (1) to investigate if transient receptor potential vanilloid‐3 (TRPV3) channels are present in mouse ESCs (mESCs) and their subcellular localization; (2) to investigate the role of TRPV3 in maintaining the characteristics of mESCs. Western blot and immunocytochemistry showed that TRPV3 was present at the endoplasmic reticulum (ER) of mESCs. Calcium imaging showed that, in the absence of extracellular calcium, TRPV3 activators camphor and 6‐tert‐butyl‐ m ‐cresol increased the cytosolic calcium. However, depleting the ER store in advance of activator addition abolished the calcium increase, suggesting that TRPV3 released calcium from the ER. To dissect the functional role of TRPV3, TRPV3 was activated and mESC proliferation was measured by trypan blue exclusion and MTT assays. The results showed that TRPV3 activation led to a decrease in mESC proliferation. Cell cycle analysis revealed that TRPV3 activation increased the percentage of cells in G 2 /M phase; consistently, Western blot also revealed a concomitant increase in the expression of inactive form of cyclin‐dependent kinase 1, suggesting that TRPV3 activation arrested mESCs at G 2 /M phase. TRPV3 activation did not alter the expression of pluripotency markers Oct‐4, Klf4 and c‐Myc, suggesting that the pluripotency was preserved. Our study is the first study to show the presence of TRPV3 at ER. Our study also reveals the novel role of TRPV3 in controlling the cell cycle and preserving the pluripotency of ESCs. J. Cell. Physiol. 231: 403–413, 2016. © 2015 Wiley Periodicals, Inc.