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001-es BibID:BIBFORM017084
Első szerző:Oláh Tamás (élettanász)
Cím:The Alterations of Store-Operated Calcium Entry in TRPC1-Overexpressing C2C12 Myotubes / Tamás Oláh, János Fodor, Olga Ruzsnavszky, Celine Berbey, Bruno Allard, László Csernoch
Dátum:2010
ISSN:0006-3495
Megjegyzések:When the endoplasmic reticulum (ER) calcium store is depleted, a Ca2+ influx is activated from the extracellular milieu to refill the intracellular stores. This well-regulated Ca2+ uptake mechanism, called store-operated Ca2+ entry (SOCE), depends on the cooperation of several proteins as STIM1, Orai1 and TRPC1. The role of STIM1 as the calcium sensor of the ER and Orai1 as the Ca2+ influx channel is well-known from the recent publications, but the function of TRPC1 as a store-operated channel remains elusive.Here TRPC1 was overexpressed by liposome-mediated transfection in C2C12 mouse skeletal muscle cell line. Overexpression was confirmed at mRNA level by RT-PCR and at protein level by immunostaining and Western-blot. The SOCE mechanism was studied by measuring the changes in [Ca2+]i evoked by the re-addition of 1.8 mM [Ca2+]e following the SERCA-inhibition by thapsigargin. As a result of TRPC1 overexpression, the amplitude and the maximum of the derivative of SOCE was significantly increased. When YM-58483, the antagonist of TRPC1 was used, these differences were eliminated, moreover in TRPC1-overexpressing myotubes the SOCE was slightly but not significantly lower, suggesting the downregulation of the STIM1-Orai1 system. This decrease in the expression level of STIM1 was confirmed by Western-blot together with the downregulation of SERCA. As a consequence a reduction in maximal Ca2+ uptake, and a higher resting [Ca2+]i following the transients evoked by 120 mM KCl were detected. Morphological changes caused by the overexpression of TRPC1 were also observed. The differentiation of the myoblasts started later, and the myotubes were thinner in TRPC1-overexpressing cultures.Our results suggest that enhancing the expression level of TRPC1 increases SOCE and has a negative feedback effect on the STIM1-Orai1 system, suggesting a cooperation between these proteins.
Tárgyszavak:Természettudományok Biológiai tudományok idézhető absztrakt
Megjelenés:Biophysical Journal. - 98 : 3 Suppl. 1 (2010), p. 152a-153a. -
További szerzők:Fodor János (1973-) (élettanász, biotechnológus) Ruzsnavszky Olga (1983-) (élettanász) Berbey, Celine Allard, Bruno Csernoch László (1961-) (élettanász)
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