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001-es BibID:	BIBFORM062410
Első szerző:	Tóth Adrienn (molekuláris biológus, élettanász)
Cím:	Investigation of the role of SERCA1b in the calcium homeostasis of C2C12 skeletal muscle cells / Tóth Adrienn, Fodor János, Vincze János, Oláh Tamás, Juhász Tamás, Zádor Ernő, Csernoch László
Dátum:	2015
Megjegyzések:	The neonatal isoform of the sarcoplasmic/endoplasmic reticulum Ca2+ ATPase 1 (SERCA1b) is a dominant Ca2+ pump in fibers of regenerating muscle. Our aim was to study the role of SERCA1b during skeletal muscle differentiation and to provide a detailed understanding of role in Ca2+ homeostasis. SERCA1b protein synthesis has been suppressed using a specific shRNA sequence. Clones with decreased SERCA 1b expression were selected for additional experiments. Scrambled shRNA transfected and parental cells were used as controls. Of the main regulatory proteins, expressions of phosphoprotein kinase B (P-Akt), myogenic differentiation factor (MyoD), stromal interaction molecule 1 (STIM1), calsequestrin (CSQ), and calcineurin were confirmed at the protein level, while myostatin and myocyte-enriched calcineurin-interacting protein (MCIP1.4) were identified at mRNA level. Quantitative analysis of the Western blots normalized to actin also confirmed the significant alterations in CSQ, STIM1, P-Akt, and calcineurin expression detected in the clones as compared to control. Furthermore, to examine the functional consequences of the decreased expression of SERCA1b, repeated Ca2+-transients were evoked by the applications of 120 mM KCl. Significantly higher [Ca2+]i was measured in the 20th and 40th seconds after the beginning of KCl application (112.3 ? 3.2 nM, and 110.4 ? 3.1 vs. 150.3 ? 6.5 nM and 134.9 ? 4.7, in control and in the clone C1 respectively) indicating decreased Ca2+ uptake capability of the SERCA pumps. This maximal pump rate was 453.6 ? 41.01 vs. 143.9 ? 24.01 lM/s, in control and in the clone C1. These results suggest that SERCA1b plays an essential role in the regulation of [Ca2+]i and ab ovo gene silencing results in decreased skeletal muscle differentiation. This work was supported by Grants: OTKA NN-107765, TÁMOP-4.2.1/B-09/1/KONV-2010-0007 and TÁMOP-4.2.2/B-10/1-2010-0024, Bólyai Research Scholarship of the Hungarian Academy of Sciences to J.F.
Tárgyszavak:	Orvostudományok Elméleti orvostudományok idézhető absztrakt C2C12 cell SERCA1b Calcium homeostasis Molekuláris Medicina Doktori iskola
Megjelenés:	Journal of muscle research and cell motility. - 36 (2015), p. 132. -
További szerzők:	Fodor János (1973-) (élettanász, biotechnológus) Vincze János (1947-) (biofizikus) Oláh Tamás (1983-) (élettanász) Juhász Tamás (1976-) (biológus, orvosbiológus) Zádor Ernő (1954-) Csernoch László (1961-) (élettanász)
Pályázati támogatás:	NN-107765 OTKA TÁMOP-4.2.1/B-09/1/KONV-2010-0007 TÁMOP A harántcsíkolt izom kontrakció molekuláris szabályozása TÁMOP-4.2.2/B-10/1-2010-0024 TÁMOP Molekuláris Orvostudomány Doktori Iskola Bólyai Research Scholarship of the Hungarian Academy of Sciences to J.F Egyéb
Internet cím:	DOI Intézményi repozitóriumban (DEA) tárolt változat
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