Stable expression and coupling of cardiac L-type Ca2+ channels with β1-adrenoceptors

A. Yatani, M. Wakamori, T. Niidome, S. Yamamoto, I. Tanaka, Y. Mori, K. Katayama, S. Green

Research output: Contribution to journalArticlepeer-review

29 Citations (Scopus)

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 sub-types in native cells. To study the functional coupling of a particular receptor subtype to these channels, rabbit cardiac Ca2+ channel α1 and skeletal β and α2/δ 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 β1-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.

Original languageEnglish
Pages (from-to)335-342
Number of pages8
JournalCirculation research
Volume76
Issue number3
Publication statusPublished - 1995 Mar 1
Externally publishedYes

Keywords

  • Ca channels
  • baby hamster kidney cells
  • patch clamp
  • β-adrenoceptors

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

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