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J Biol Chem, Vol. 275, Issue 20, 15239-15245, May 19, 2000

Ca2+/Calmodulin-dependent Kinase II and Calcineurin Play Critical Roles in Endothelin-1-induced Cardiomyocyte Hypertrophy*

Weidong ZhuDagger , Yunzeng ZouDagger , Ichiro ShiojimaDagger , Sumiyo KudohDagger , Ruichi AikawaDagger , Dobun HayashiDagger , Miho MizukamiDagger , Haruhiro TokoDagger , Futoshi Shibasaki§, Yoshio YazakiDagger , Ryozo NagaiDagger , and Issei KomuroDagger

From the Dagger  Department of Cardiovascular Medicine, University of Tokyo Graduate School of Medicine, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan and § Tokyo Metropolitan Institute of Medical Science, Tokyo 113-8655, Japan

Endothelin-1 (ET-1) induces cardiac hypertrophy. Because Ca2+ is a major second messenger of ET-1, the role of Ca2+ in ET-1-induced hypertrophic responses in cultured cardiac myocytes of neonatal rats was examined. ET-1 activated the promoter of the beta -type myosin heavy chain gene (beta -MHC) (-354 to +34 base pairs) by about 4-fold. This activation was inhibited by chelation of Ca2+ and the blocking of protein kinase C activity. Similarly, the beta -MHC promoter was activated by Ca2+ ionophores and a protein kinase C activator. beta -MHC promoter activation induced by ET-1 was suppressed by pretreatment with the calmodulin inhibitor, W7, the Ca2+/calmodulin-dependent kinase II (CaMKII) inhibitor, KN62, and the calcineurin inhibitor, cyclosporin A. beta -MHC promoter activation by ET-1 was also attenuated by overexpression of dominant-negative mutants of CaMKII and calcineurin. ET-1 increased the activity of CaMKII and calcineurin in cardiac myocytes. Pretreatment with KN62 and cyclosporin A strongly suppressed ET-1-induced increases in [3H]phenylalanine uptake and in cell size. These results suggest that Ca2+ plays a critical role in ET-1-induced cardiomyocyte hypertrophy by activating CaMKII- and calcineurin-dependent pathways.

* This work was supported by a Grant-in-Aid for Scientific Research, Developmental Scientific Research and Scientific Research on Priority Areas from the Ministry of Education, Science, Sports, and Culture of Japan and by Program for Promotion of Fundamental Studies in Health Sciences of the Organization for Drug ADR Relief, R&D Promotion, and Product Review of Japan (to I. K.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed. Tel.: 81-3-3818-6672; Fax: 81-3-3818-6673; E-mail: komuro-tky@umin.ac.jp.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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