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(Circulation Research. 1995;76:335-342.)
© 1995 American Heart Association, Inc.

Articles

Stable Expression and Coupling of Cardiac L-Type Ca²⁺ Channels With ß₁-Adrenoceptors

Atsuko Yatani, Minoru Wakamori, Tetsuhiro Niidome, Satoshi Yamamoto, Isao Tanaka, Yasuo Mori, Kouich Katayama, Stuart Green

From the Departments of Pharmacology and Cell Biophysics (A.Y., M.W., S.Y., Y.M.) and Medicine (Pulmonary) (A.Y., S.G.), University of Cincinnati (Ohio) College of Medicine, and Tsukuba Research Laboratories (T.N., I.T., K.K.), Eisai Co Ltd, Ibaraki, Japan.

Correspondence to Dr A. Yatani, Department of Pharmacology and Cell Biophysics, University of Cincinnati College of Medicine, Cincinnati, OH 45267-0575.

Abstract A number of neurotransmitters modulate cardiac dihydropyridine-sensitive L-type Ca²⁺ channels through several homologous G protein–coupled receptors. Previous studies that have examined receptor–Ca²⁺ channel interactions have suffered because of the coexpression of various receptor subtypes in native cells. To study the functional coupling of a particular receptor subtype to these channels, rabbit cardiac Ca²⁺ channel ₁ and skeletal ß and ₂/ subunits were stably expressed in baby hamster kidney cells. In this stable cell line, Ca²⁺ channels remained at high levels (>1000 fmol/mg protein, or 2700 channels per cell) over extended times. The expressed recombinant Ca²⁺ channels displayed the voltage dependence of activation and inactivation, unitary conductance, and pharmacology characteristic of native cardiac L-type Ca²⁺ channels. Subsequent coexpression of the ß₁-adrenoceptors (150 to 300 fmol/mg protein) with the Ca²⁺ channels resulted in cell responsiveness to the extracellular application of isoproterenol. These results indicate that heterogeneous expression in mammalian cells provides a useful system for studying both biophysical analysis of Ca²⁺ channel properties and receptor-coupled regulatory processes.

Key Words: Ca²⁺ channels • ß₁-adrenoceptors • patch clamp • baby hamster kidney cells

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