Characterization of voltage-dependent calcium channels expressed in Xenopus oocytes injected with mRNA from rat heart

J Physiol. 1990 Oct:429:95-112. doi: 10.1113/jphysiol.1990.sp018246.

Abstract

1. The properties of voltage dependent cardiac Ca channels expressed in Xenopus laevis oocytes after injection of mRNA from rat heart were investigated using the double-microelectrode voltage-clamp technique. 2. Endogenous Ba current (IBa,E) and expressed cardiac Ba current (IBa,C) were studied at various external concentrations of barium (Ba2+). These two entities could be distinguished by their amplitude and their pharmacology. IBa,C was more sensitive to the inorganic Ca channel blocker manganese (Mn2+). The contaminant IBa,E presented properties of voltage dependence identical to IBa,C, but was negligible in the presence of a low external Ba2+ concentration (2 mM). 3. In 2 mM-Ba2+, IBa,C activated at -35 mV, peaked at -14 mV, and reversed at +26 mV. Steady-state inactivation properties, in consideration of the half-inactivation potential of -35 mV, were also typical of L-type Ba currents. However, the decay of IBa,C was very slow (time constant of inactivation near 600 ms). No evidence for the expression of cardiac transient Ca channels (T-type) was found. 4. IBa,C was enhanced after exposure to the 1,4-dihydropyridine (DHP) agonist Bay K 8644. The enhancement of IBa,C was voltage dependent (maximum at -30 +/- 5 mV) and associated with a slowing in current decay. Current-voltage and concentration-response curves obtained for various Ba2+ concentrations revealed an antagonism between external Ba2+ and the 1,4-DHP agonist Bay K 8644. Similar results were found using the (-)Bay K 8644 pure agonist isomer. 5. We conclude that oocytes injected with mRNA from rat heart expressed only the high threshold, long-lasting or L-type Ca channels. The availability of expressed L-type Ca channels for quantitative pharmacological studies using low Ba2+ concentration has been demonstrated.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / pharmacology
  • Animals
  • Barium / pharmacology
  • Cadmium / pharmacology
  • Calcium Channel Blockers / pharmacology
  • Calcium Channels / drug effects
  • Calcium Channels / physiology*
  • Female
  • In Vitro Techniques
  • Manganese / pharmacology
  • Membrane Potentials / drug effects
  • Oocytes / physiology*
  • RNA, Messenger / metabolism*
  • Xenopus laevis

Substances

  • Calcium Channel Blockers
  • Calcium Channels
  • RNA, Messenger
  • Cadmium
  • Barium
  • Manganese
  • 3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester