Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
CAAS-ILRI Joint Laboratory on Livestock and Forage Genetic Resources, Institute of Animal Science, Chinese Academy of Agricultural Sciences (CAAS), Beijing 100193, China.
Int J Mol Sci. 2022 Dec 16;23(24):16065. doi: 10.3390/ijms232416065.
As relatively new members of the non-coding RNA family, circRNAs play important roles in a variety of biological processes. However, the temporal expression pattern and the function of circRNAs during sheep skeletal muscle development remains unclear. This study aimed to identify circRNAs related to sheep skeletal muscle development and explore their roles in myoblast proliferation. The circRNA expression profiles of longissimus dorsi of sheep from F90, L30, and A3Y were obtained by the RNA-seq method. The function and mechanisms of the novel circCHRNG in muscle satellite cell proliferation were explored using CCK-8 assay, Western blot, qPCR, and dual-luciferase reporter assay. We identified 12,375 circRNAs, including 476, 133, and 233 DEcircRNAs found among three comparative groups. KEGG results showed that DEcircRNAs were enriched in muscle contraction, the regulation of cell proliferation, and the AMPK, insulin, and PI3K-Akt signaling pathways. Notably, a novel circRNA, termed circRNA CHRNG, acts as a miR-133 sponge to promote skeletal muscle satellite cell proliferation. Our study provides a systematic description of circRNAs of ovine skeletal muscle across fetal, lamb, and adult stages. GO and KEGG analyses showed that DEcircRNAs were enriched in multiple pathways associated with muscle development, such as the PI3K-Akt and AMPK signaling pathways. In addition, we propose that circCHRNG acts as a miR-133 sponge to upregulate the expression levels of SRF and MEF2A, thereby promoting myoblast proliferation.
作为非编码 RNA 家族的相对较新成员，circRNAs 在多种生物过程中发挥着重要作用。然而，circRNAs 在绵羊骨骼肌发育过程中的时空表达模式和功能仍不清楚。本研究旨在鉴定与绵羊骨骼肌发育相关的 circRNAs，并探索其在成肌细胞增殖中的作用。通过 RNA-seq 方法获得了 F90、L30 和 A3Y 绵羊背最长肌的 circRNA 表达谱。使用 CCK-8 测定法、Western blot、qPCR 和双荧光素酶报告基因测定法探索了新型 circCHRNG 在肌肉卫星细胞增殖中的功能和机制。我们鉴定了 12375 个 circRNAs，其中包括在三个比较组中发现的 476、133 和 233 个差异表达 circRNAs。KEGG 结果表明，差异表达 circRNAs 富集在肌肉收缩、细胞增殖的调控以及 AMPK、胰岛素和 PI3K-Akt 信号通路中。值得注意的是，一种新型 circRNA，称为 circRNA CHRNG，作为 miR-133 的海绵体促进骨骼肌卫星细胞增殖。本研究提供了绵羊骨骼肌在胎儿、羔羊和成年阶段的 circRNAs 的系统描述。GO 和 KEGG 分析表明，差异表达 circRNAs 富集在与肌肉发育相关的多个通路中，如 PI3K-Akt 和 AMPK 信号通路。此外，我们提出 circCHRNG 作为 miR-133 的海绵体，上调 SRF 和 MEF2A 的表达水平，从而促进成肌细胞增殖。
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