Alterations of excitation–contraction coupling and excitation coupled Ca2+ entry in human myotubes carrying CAV3 mutations linked to rippling muscle

✍ Scribed by Nina D. Ullrich; Dirk Fischer; Cornelia Kornblum; Maggie C. Walter; Ernst Niggli; Francesco Zorzato; Susan Treves

• Publisher: John Wiley and Sons
• Year: 2011
• Tongue: English
• Weight: 370 KB
• Volume: 32
• Category: Article
• ISSN: 1059-7794
• DOI: 10.1002/humu.21431

✦ Synopsis

Rippling muscle disease is caused by mutations in the gene encoding caveolin-3 (CAV3), the muscle-specific isoform of the scaffolding protein caveolin, a protein involved in the formation of caveolae. In healthy muscle, caveolin-3 is responsible for the formation of caveolae, which are highly organized sarcolemmal clusters influencing early muscle differentiation, signalling and Ca^2+^ homeostasis. In the present study we examined Ca^2+^ homeostasis and excitation–contraction (E-C) coupling in cultured myotubes derived from two patients with Rippling muscle disease with severe reduction in caveolin-3 expression; one patient harboured the heterozygous c.84C>A mutation while the other patient harbored a homozygous splice-site mutation (c.102+ 2T>C) affecting the splice donor site of intron 1 of the CAV3 gene. Our results show that cells from control and rippling muscle disease patients had similar resting [Ca^2+^]~i~ and 4-chloro-m-cresol-induced Ca^2+^ release but reduced KCl-induced Ca^2+^ influx. Detailed analysis of the voltage-dependence of Ca^2+^ transients revealed a significant shift of Ca^2+^ release activation to higher depolarization levels in CAV3 mutated cells. High resolution immunofluorescence analysis by Total Internal Fluorescence microscopy supports the hypothesis that loss of caveolin-3 leads to microscopic disarrays in the colocalization of the voltage-sensing dihydropyridine receptor and the ryanodine receptor, thereby reducing the efficiency of excitation–contraction coupling. Hum Mutat 32:309–317, 2011. © 2011 Wiley-Liss, Inc.