Increased store-operated Ca2+ entry in skeletal muscle with reduced calsequestrin-1 expression.
Store-operated Ca(2+) entry (SOCE) contributes to Ca(2+) handling in normal skeletal muscle function, as well as the progression of muscular dystrophy and sarcopenia, yet the mechanisms underlying the change in SOCE in these states remain unclear. Previously we showed that calsequestrin-1 (CSQ1) participated in retrograde regulation of SOCE in cultured skeletal myotubes. In this study, we used small-hairpin RNA to determine whether knockdown of CSQ1 in adult mouse skeletal muscle can influence SOCE activity and muscle function. Small-hairpin RNA against CSQ1 was introduced into flexor digitorum brevis muscles using electroporation. Transfected fibers were isolated for SOCE measurements using the Mn(2+) fluorescence-quenching method. At room temperature, the SOCE induced by submaximal depletion of the SR Ca(2+) store was significantly enhanced in CSQ1-knockdown muscle fibers. When temperature of the bathing solution was increased to 39 degrees C, CSQ1-knockdown muscle fibers displayed a significant increase in Ca(2+) permeability across the surface membrane likely via the SOCE pathway, and a corresponding elevation in cytosolic Ca(2+) as compared to control fibers. Preincubation with azumolene, an analog of dantrolene used for the treatment of malignant hyperthermia (MH), suppressed the elevated SOCE in CSQ1-knockdown fibers. Because the CSQ1-knockout mice develop similar MH phenotypes, this inhibitory effect of azumolene on SOCE suggests that elevated extracellular Ca(2+) entry in skeletal muscle may be a key factor for the pathophysiological changes in intracellular Ca(2+) signaling in MH.
Zhao
Xiaoli
X
Department of Physiology and Biophysics, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway, New Jersey, USA.
Min
Choon Kee
CK
Ko
Jae-Kyun
JK
Parness
Jerome
J
Kim
Do Han
DH
Weisleder
Noah
N
Ma
Jianjie
J
D000818:Animals; D001692:Biological Transport / Q000235:genetics; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000172:deficiency* / Q000235:genetics* / Q000378:metabolism; D002155:Calsequestrin; D005110:Extracellular Space / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000235:genetics*; D055785:Gene Knockdown Techniques; D057809:HEK293 Cells; D018869:Heat-Shock Response / Q000187:drug effects; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D055029:Inverted Repeat Sequences; D008305:Malignant Hyperthermia / Q000235:genetics / Q000378:metabolism; D051379:Mice; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010080:Oxazoles / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000235:genetics; D012519:Sarcoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism
D002135:Calcium-Binding Proteins; D002155:Calsequestrin; C503130:Casq1 protein, mouse; D007093:Imidazoles; D010080:Oxazoles; D034741:RNA, Small Interfering; C061387:azumolene; D002118:Calcium
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