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Am J Physiol Cell Physiol (April 30, 2003). doi:10.1152/ajpcell.00072.2003
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Submitted on February 21, 2003
Accepted on April 22, 2003

Ca2+ Spark Occurrence In Embryonic Mouse Skeletal Muscle Is Initiated By Ca2+ Entry And Declines During Postnatal Development

Lois G Chun1, Christopher W Ward2, and Martin F Schneider1*

1 Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, MD, USA
2 Department of Adult Health Nursing, University of Maryland School of Nursing, Baltimore, MD, USA; Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, MD, USA

* To whom correspondence should be addressed. E-mail: mschneid{at}umaryland.edu.

"Spontaneous" Ca2+ sparks and ryanodine receptor type 3 (RyR3) expression are readily detected in embryonic mammalian skeletal muscle but not in adult mammalian muscle, which rarely exhibits Ca2+ sparks and expresses predominantly RyR1. Here we use confocal fluorescence imaging and systematic sampling of enzymatically dissociated single striated muscle fibers containing the Ca2+ indicator dye Fluo-4 to show that the frequency of spontaneous Ca2+ sparks decreases dramatically from embryonic day 18 (E18) to postnatal day 14 (P14) in mouse diaphragm and from P1 to P14 in mouse EDL fibers. In contrast, the relative levels of RyR3 to RyR1 protein remained constant in diaphragm muscles from E18 to P14, indicating that changes in relative levels of RyR isoform expression did not cause the decline in Ca2+ spark frequency. E18 diaphragm fibers were used to investigate possible mechanisms underlying spark initiation in embryonic fibers. Spark frequency increased or decreased, respectively, when E18 diaphragm fibers were exposed to 8 or 0 mM Ca2+ in the extracellular Ringer's solution, with no change in either the average resting fiber Fluo-4 fluorescence or the average properties of the sparks. Either CoCl2 (5 mM) or nifedipine (30 µM) markedly decreased spark frequency in E18 diaphragm fibers. These results indicate that Ca2+ sparks may be triggered by locally elevated [Ca2+] due to Ca2+ influx via dihydropyridine receptor (DHPR) L-type Ca2+ channels in embryonic mammalian skeletal muscle.




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