title: Temperature and RyR1 regulate the activation rate of store-operated Ca²+ entry current in myotubes.
abstract: Store-operated calcium entry (SOCE) is an important Ca(2+) entry pathway in skeletal muscle. However, direct electrophysiological recording and full characterization of the underlying SOCE current in skeletal muscle cells (I(SkCRAC)) has not been reported. Here, we characterized the biophysical properties, pharmacological profile, and molecular identity of I(SkCRAC) in skeletal myotubes, as well as the regulation of its rate of activation by temperature and the type I ryanodine receptor (RyR1). I(SkCRAC) exhibited many hallmarks of Ca(2+) release activated Ca(2+) currents (I(CRAC)): store dependence, strong inward rectification, positive reversal potential, limited cesium permeability, and sensitivity to SOCE channel blockers. I(SkCRAC) was reduced by siRNA knockdown of stromal interaction molecule 1 and expression of dominant negative Orai1. Average I(SkCRAC) current density at -80mV was 1.00 ± 0.05 pA/pF. In the presence of 20 mM intracellular EGTA, I(SkCRAC) activation occurred over tens of seconds during repetitive depolarization at 0.5Hz and was inhibited by treatment with 100 μM ryanodine. The rate of SOCE activation was reduced threefold in myotubes from RyR1-null mice and increased 4.6-fold at physiological temperatures (35-37°C). These results show that I(SkCRAC) exhibits similar biophysical, pharmacological, and molecular properties as I(CRAC) in nonexcitable cells and its rate of activation during repetitive depolarization is strongly regulated by temperature and RyR1 activity.
authors.affiliation: Department of Pharmacology and Physiology, University of Rochester Medical Center, Rochester, New York, USA.
authors.forename: Viktor
authors.initials: V
authors.lastname: Yarotskyy
authors.forename: Robert T
authors.initials: RT
authors.lastname: Dirksen
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism; D015640:Ion Channel Gating; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D018485:Muscle Fibers, Skeletal / Q000378:metabolism*; D000071740:ORAI1 Protein; D019837:Ryanodine Receptor Calcium Release Channel / Q000378:metabolism*; D000071737:Stromal Interaction Molecule 1; D013696:Temperature; D013997:Time Factors
chemical_list: D015220:Calcium Channels; D008562:Membrane Glycoproteins; D000071740:ORAI1 Protein; C520430:Orai1 protein, mouse; D019837:Ryanodine Receptor Calcium Release Channel; C100757:Stim1 protein, mouse; D000071737:Stromal Interaction Molecule 1; D002118:Calcium