RRC ID 57979
著者 Hashimoto M, Saito N, Ohta H, Yamamoto K, Tashiro A, Nakazawa K, Inanami O, Kitamura H.
タイトル Inhibition of ubiquitin-specific protease 2 causes accumulation of reactive oxygen species, mitochondria dysfunction, and intracellular ATP decrement in C2C12 myoblasts.
ジャーナル Physiol Rep
Abstract Ubiquitin-specific protease 2 (USP2) is considered to participate in the differentiation of myoblasts to myotubes, however, its functions in myoblasts under growth conditions remain elusive. In this study, we analyzed the physiological roles of USP2 in myoblasts using Usp2 knockout (KO) C2C12 cells as well as a USP2 specific inhibitor. In addition to the disruption of differentiation, clustered regularly interspaced short palindromic repeats/Cas9-generated Usp2KO cells exhibited inhibition of proliferation compared to parental C2C12 cells. Usp2KO cells reduced the accumulation of intracellular adenosine triphosphate (ATP) content and oxygen consumption. Moreover, Usp2KO cells had fragmented mitochondria, suggesting that mitochondrial respiration was inactive. The deficiency of Usp2 did not affect the enzymatic activities of respiratory chain complexes I, III, IV, and V. However, mitochondrial membrane permeability-evaluated using calcein AM-cobalt staining-was increased in Usp2KO cells. The membrane potential of Usp2KO cells was clearly decreased. Usp2KO cells accumulated reactive oxygen species (ROS) in the mitochondria. The USP2-selective inhibitor ML364 also increased the levels of mitochondrial ROS, and modulated the membrane potential and morphology of the mitochondria. These effects were followed by a decrement in the intracellular content of ATP. Based on these findings, we speculate that USP2 may be involved in maintaining the integrity of the mitochondrial membrane. This process ensures the supply of ATP in myoblasts, presumably leading to proliferation and differentiation.
巻・号 7(14)
ページ e14193
公開日 2019-7-1
DOI 10.14814/phy2.14193
PMID 31353872
PMC PMC6661303
MeSH Adenosine Triphosphate / metabolism* Animals Cell Line Enzyme Inhibitors / pharmacology Membrane Potential, Mitochondrial Mice Mitochondria, Muscle / drug effects Mitochondria, Muscle / metabolism* Myoblasts / drug effects Myoblasts / metabolism Oxidative Phosphorylation Reactive Oxygen Species / metabolism* Ubiquitin Thiolesterase / antagonists & inhibitors Ubiquitin Thiolesterase / genetics Ubiquitin Thiolesterase / metabolism*
引用数 2
リソース情報
ヒト・動物細胞 C2C12(RCB0987)