Cardiac-type EC-coupling in dysgenic myotubes restored with Ca2+ channel subunit isoforms alpha1C and alpha1D does not correlate with current density

Biophys J. 2003 Jun;84(6):3816-28. doi: 10.1016/S0006-3495(03)75109-1.

Abstract

Ca(2+)-induced Ca(2+)-release (CICR)-the mechanism of cardiac excitation-contraction (EC) coupling-also contributes to skeletal muscle contraction; however, its properties are still poorly understood. CICR in skeletal muscle can be induced independently of direct, calcium-independent activation of sarcoplasmic reticulum Ca(2+) release, by reconstituting dysgenic myotubes with the cardiac Ca(2+) channel alpha(1C) (Ca(V)1.2) subunit. Ca(2+) influx through alpha(1C) provides the trigger for opening the sarcoplasmic reticulum Ca(2+) release channels. Here we show that also the Ca(2+) channel alpha(1D) isoform (Ca(V)1.3) can restore cardiac-type EC-coupling. GFP-alpha(1D) expressed in dysgenic myotubes is correctly targeted into the triad junctions and generates action potential-induced Ca(2+) transients with the same efficiency as GFP-alpha(1C) despite threefold smaller Ca(2+) currents. In contrast, GFP-alpha(1A), which generates large currents but is not targeted into triads, rarely restores action potential-induced Ca(2+) transients. Thus, cardiac-type EC-coupling in skeletal myotubes depends primarily on the correct targeting of the voltage-gated Ca(2+) channels and less on their current size. Combined patch-clamp/fluo-4 Ca(2+) recordings revealed that the induction of Ca(2+) transients and their maximal amplitudes are independent of the different current densities of GFP-alpha(1C) and GFP-alpha(1D). These properties of cardiac-type EC-coupling in dysgenic myotubes are consistent with a CICR mechanism under the control of local Ca(2+) gradients in the triad junctions.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Action Potentials / physiology*
  • Adaptation, Physiological / physiology
  • Calcium / physiology
  • Calcium Channels
  • Calcium Channels, L-Type / physiology*
  • Calcium Signaling / physiology*
  • Cell Line
  • Heart / physiology*
  • Membrane Potentials / physiology*
  • Muscle Contraction / physiology*
  • Muscle Fibers, Skeletal
  • Muscle, Skeletal / physiology*
  • Mutagenesis, Site-Directed
  • Mutation
  • Myocardial Contraction / physiology
  • Protein Isoforms
  • Protein Subunits
  • Recombinant Proteins / metabolism
  • Sarcoplasmic Reticulum / physiology
  • Statistics as Topic
  • Structure-Activity Relationship

Substances

  • Calcium Channels
  • Calcium Channels, L-Type
  • L-type calcium channel alpha(1C)
  • Protein Isoforms
  • Protein Subunits
  • Recombinant Proteins
  • Cacna1d protein, rat
  • Calcium