期刊
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
卷 532, 期 3, 页码 482-488出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bbrc.2020.08.080
关键词
C2C12 myoblasts; Chloride channel; Intracellular chloride; Myogenesis; Volume-regulated anion channel
资金
- German Federal Ministry of Education and Research (BMBF) [031A314]
- Oversea Study Program of Guangzhou Elite Project [JY201624]
Leucine-rich repeat containing family 8 (LRRC8) proteins form the volume-regulated anion channel (VRAC). Recently, they were shown to be required for normal differentiation and fusion of C2C12 myoblasts, by promoting membrane hyperpolarization and intracellular Ca2+ signals. However, the mechanism by which they are involved remained obscure. Here, using a FRET-based sensor for VRAC activity, we show temporary activation of VRAC within the first 2 h of myogenic differentiation. During this period, we also observed a significant decrease in the intracellular Cl- concentration that was abolished by the VRAC inhibitor carbenoxolone. However, lowering the intracellular Cl(- )concentration by extracellular Cl(- )depletion did not promote differentiation as judged by the percentage of myogenin- positive nuclei or total myogenin levels in C2C12 cells. Instead, it inhibited myosin expression and myotube formation. Together, these data suggest that VRAC is activated and mediates Cl(- )efflux early on during myogenic differentiation, and a moderate intracellular Cl(- )concentration is necessary for myoblast fusion. (C) 2020 The Authors. Published by Elsevier Inc.
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