Altered expression of triadin 95 causes parallel changes in localized Ca2+ release events and global Ca2+ signals in skeletal muscle cells in culture

János Fodor, M. Gönczi, Monika Sztretye, Beatrix Dienes, Tamás Oláh, László Szabó, Eszter Csoma, P. Szentesi, G. Szigeti, Isabelle Marty, L. Csernoch

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The 95 kDa triadin (Trisk 95), an integral protein of the sarcoplasmic reticular membrane in skeletal muscle, interacts with both the ryanodine receptor (RyR) and calsequestrin. While its role in the regulation of calcium homeostasis has been extensively studied, data are not available on whether the overexpression or the interference with the expression of Trisk 95 would affect calcium sparks the localized events of calcium release (LCRE). In the present study LCRE and calcium transients were studied using laser scanning confocal microscopy on C2C12 cells and on primary cultures of skeletal muscle. Liposome- or adenovirus-mediated Trisk 95 overexpression and shRNA interference with triadin translation were used to modify the level of the protein. Stable overexpression in C2C12 cells significantly decreased the amplitude and frequency of calcium sparks, and the frequency of embers. In line with these observations, depolarization-evoked calcium transients were also suppressed. Similarly, adenoviral transfection of Trisk 95 into cultured mouse skeletal muscle cells significantly decreased both the frequency and amplitude of spontaneous global calcium transients. Inhibition of endogenous triadin expression by RNA interference caused opposite effects. Primary cultures of rat skeletal muscle cells expressing endogenous Trisk 95 readily generated spontaneous calcium transients but rarely produced calcium sparks. Their transfection with specific shRNA sequence significantly reduced the triadin-specific immunoreactivity. Functional experiments on these cells revealed that while caffeine-evoked calcium transients were reduced, LCRE appeared with higher frequency. These results suggest that Trisk 95 negatively regulates RyR function by suppressing localized calcium release events and global calcium signals in cultured muscle cells.

Original languageEnglish
Pages (from-to)5803-5818
Number of pages16
JournalJournal of Physiology
Volume586
Issue number23
DOIs
Publication statusPublished - Dec 2008

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Muscle Cells
Skeletal Muscle
Cell Culture Techniques
Calcium
Calcium Signaling
Ryanodine Receptor Calcium Release Channel
Small Interfering RNA
Transfection
triadin
Calsequestrin
Primary Cell Culture
RNA Interference
Caffeine
Adenoviridae
Confocal Microscopy
Liposomes
Cultured Cells
Proteins
Homeostasis
Membranes

ASJC Scopus subject areas

  • Physiology

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Altered expression of triadin 95 causes parallel changes in localized Ca2+ release events and global Ca2+ signals in skeletal muscle cells in culture. / Fodor, János; Gönczi, M.; Sztretye, Monika; Dienes, Beatrix; Oláh, Tamás; Szabó, László; Csoma, Eszter; Szentesi, P.; Szigeti, G.; Marty, Isabelle; Csernoch, L.

In: Journal of Physiology, Vol. 586, No. 23, 12.2008, p. 5803-5818.

Research output: Contribution to journalArticle

Fodor, János ; Gönczi, M. ; Sztretye, Monika ; Dienes, Beatrix ; Oláh, Tamás ; Szabó, László ; Csoma, Eszter ; Szentesi, P. ; Szigeti, G. ; Marty, Isabelle ; Csernoch, L. / Altered expression of triadin 95 causes parallel changes in localized Ca2+ release events and global Ca2+ signals in skeletal muscle cells in culture. In: Journal of Physiology. 2008 ; Vol. 586, No. 23. pp. 5803-5818.
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