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L 型氨基酸转运蛋白 1()在体外肌发生过程中的作用。

The role of L-type amino acid transporter 1 () during in vitro myogenesis.

机构信息

School of Human Kinetics, University of Ottawa, Ottawa, Ontario, Canada.

Department of Kinesiology and Community Health, University of Illinois at Urbana-Champaign, Urbana, Illinois.

出版信息

Am J Physiol Cell Physiol. 2022 Aug 1;323(2):C595-C605. doi: 10.1152/ajpcell.00162.2021. Epub 2022 Jul 18.

DOI:10.1152/ajpcell.00162.2021
PMID:35848618
Abstract

Satellite cells are required for muscle regeneration, remodeling, and repair through their activation, proliferation, and differentiation; however, how dietary factors regulate this process remains poorly understood. The L-type amino acid transporter 1 (LAT1) transports amino acids, such as leucine, into mature myofibers, which then stimulate protein synthesis and anabolic signaling. However, whether LAT1 is expressed on myoblasts and is involved in regulating myogenesis is unknown. The aim of this study was to characterize the expressional and functional relevance of LAT1 during different stages of myogenesis and in response to growth and atrophic conditions in vitro. We determined that LAT1 is expressed by C2C12 and human primary myoblasts, and its gene expression is lower during differentiation ( < 0.05). Pharmacological inhibition and genetic knockdown of LAT1 impaired myoblast viability, differentiation, and fusion (all < 0.05). LAT1 protein content in C2C12 myoblasts was not significantly altered in response to different leucine concentrations in cell culture media or in two in vitro atrophy models. However, LAT1 content was decreased in myotubes under atrophic conditions in vitro ( < 0.05). These findings indicate that LAT1 is stable throughout myogenesis and in response to several in vitro conditions that induce muscle remodeling. Further, amino acid transport through LAT1 is required for normal myogenesis in vitro.

摘要

卫星细胞通过激活、增殖和分化来维持肌肉再生、重塑和修复;然而,饮食因素如何调节这一过程仍知之甚少。L 型氨基酸转运蛋白 1(LAT1)将氨基酸(如亮氨酸)转运到成熟的肌纤维中,然后刺激蛋白质合成和合成代谢信号。然而,LAT1 是否在成肌细胞上表达并参与调节肌发生尚不清楚。本研究旨在探讨 LAT1 在肌发生的不同阶段以及体外生长和萎缩条件下的表达和功能相关性。我们发现 LAT1 由 C2C12 和人原代成肌细胞表达,其基因表达在分化过程中降低(<0.05)。LAT1 的药理学抑制和基因敲低会损害成肌细胞的活力、分化和融合(均<0.05)。C2C12 成肌细胞中的 LAT1 蛋白含量在细胞培养介质中的不同亮氨酸浓度或两种体外萎缩模型中没有显著改变。然而,LAT1 含量在体外萎缩条件下的肌管中减少(<0.05)。这些发现表明,LAT1 在整个肌发生过程中以及在诱导肌肉重塑的几种体外条件下都保持稳定。此外,LAT1 介导的氨基酸转运对于体外正常的肌发生是必需的。

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