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骨骼肌细胞对不同温度的形态和分子响应。

Morphological and Molecular Responses of Skeletal Muscle Cells to Different Temperatures.

机构信息

Key Laboratory of Mariculture, Ministry of Education (KLMME), Ocean University of China, Qingdao 266003, China.

出版信息

Int J Mol Sci. 2022 Aug 29;23(17):9812. doi: 10.3390/ijms23179812.

Abstract

Temperature strongly modulates muscle development and growth in ectothermic teleosts; however, the underlying mechanisms remain largely unknown. In this study, primary cultures of skeletal muscle cells of were conducted and reared at different temperatures (21, 25, and 28 °C) in both the proliferation and differentiation stages. CCK-8, EdU, wound scratch and nuclear fusion index assays revealed that the proliferation, myogenic differentiation, and migration processes of skeletal muscle cells were significantly accelerated as the temperature raises. Based on the GO, GSEA, and WGCNA, higher temperature (28 °C) induced genes involved in HSF1 activation, DNA replication, and ECM organization processes at the proliferation stage, as well as HSF1 activation, calcium activity regulation, myogenic differentiation, and myoblast fusion, and sarcomere assembly processes at the differentiation stage. In contrast, lower temperature (21 °C) increased the expression levels of genes associated with DNA damage, DNA repair and apoptosis processes at the proliferation stage, and cytokine signaling and neutrophil degranulation processes at the differentiation stage. Additionally, we screened several hub genes regulating myogenesis processes. Our results could facilitate the understanding of the regulatory mechanism of temperature on fish skeletal muscle growth and further contribute to utilizing rational management strategies and promoting organism growth and development.

摘要

温度强烈调节着变温硬骨鱼类的肌肉发育和生长;然而,其潜在机制在很大程度上仍然未知。在这项研究中,我们进行了 的骨骼肌细胞原代培养,并在增殖和分化阶段分别在不同温度(21、25 和 28°C)下培养。CCK-8、EdU、划痕实验和核融合指数测定表明,随着温度升高,骨骼肌细胞的增殖、成肌分化和迁移过程显著加快。基于 GO、GSEA 和 WGCNA 分析,高温(28°C)在增殖阶段诱导与 HSF1 激活、DNA 复制和细胞外基质组织相关的基因,以及 HSF1 激活、钙活性调节、成肌分化和肌细胞融合以及肌节组装过程;而在分化阶段,低温(21°C)则增加了与增殖阶段 DNA 损伤、DNA 修复和细胞凋亡过程,以及分化阶段细胞因子信号和嗜中性粒细胞脱颗粒过程相关的基因表达水平。此外,我们还筛选了几个调节肌发生过程的关键基因。我们的研究结果可以促进对温度调控鱼类骨骼肌生长的机制的理解,并有助于利用合理的管理策略促进机体的生长和发育。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f056/9456278/a206c1e561c5/ijms-23-09812-g001.jpg

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