Stable expression and coupling of cardiac L-type Ca2+ channels with beta 1-adrenoceptors

Circ Res. 1995 Mar;76(3):335-42. doi: 10.1161/01.res.76.3.335.

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 alpha 1 and skeletal beta and alpha 2/delta 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 beta 1-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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bucladesine / pharmacology
  • Calcium Channels / physiology*
  • Cells, Cultured
  • Cricetinae
  • Isoproterenol / pharmacology
  • Isradipine / metabolism
  • Myocardium / metabolism*
  • Rabbits
  • Receptors, Adrenergic, beta-1 / physiology*

Substances

  • Calcium Channels
  • Receptors, Adrenergic, beta-1
  • Bucladesine
  • Isoproterenol
  • Isradipine