Caffeine-induced release of intracellular Ca2+ from chinese hamster ovary cells expressing skeletal muscle ryanodine receptor: Effects on full- length and carboxyl-terminal portion of Ca2+ release channels

Manjunatha B. Bhat, Jiying Zhao, Weijin Zang, C. William Balke, Hiroshi Takeshima, W. Gil Wier, Jianjie Ma

Research output: Contribution to journalArticle

79 Scopus citations

Abstract

The ryanodine receptor (RyR)/Ca2+ release channel is an essential component of excitation-contraction coupling in striated muscle cells. To study the function and regulation of the Ca2+ release channel, we tested the effect of caffeine on the full-length and carboxyl-terminal portion of skeletal muscle RyE expressed in a Chinese hamster ovary (CHO) cell line. Caffeine induced openings of the full length RyR channels in a concentration- dependent manner, but it had no effect on the carboxyl-terminal RyE channels. CHO cells expressing the carboxyl-terminal RyE proteins displayed spontaneous changes of intracellular [Ca2+]. Unlike the native RyE channels in muscle cells, which display localized Ca2+ release events (i.e., 'Ca2+ sparks' in cardiac muscle and 'local release events' in skeletal muscle), CHO cells expressing the full length RyE proteins did not exhibit detectable spontaneous or caffeine-induced local Ca2+ release events. Our data suggest that the binding site for caffeine is likely to reside within the amino- terminal portion of RyR, and the localized Ca2+ release events observed in muscle cells may involve gating of a group of Ca2+ release channels and/or interaction of RyE with muscle-specific proteins.

Original languageEnglish
Pages (from-to)749-762
Number of pages14
JournalJournal of General Physiology
Volume110
Issue number6
DOIs
StatePublished - Dec 1 1997
Externally publishedYes

Keywords

  • Calcium sparks
  • Chinese hamster ovary cells
  • Excitation-contraction coupling
  • Fura-2
  • Green fluorescent protein

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