Zhe Yuqi, Wu Zhijuan, Yasenjian Sibinuer, Zhong Jincheng, Jiang Hui, Zhang Ming, Chai Zhixin, Xin Jinwei
Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization, Sichuan Province and Ministry of Education, Southwest Minzu University, Chengdu, China.
Sichuan Qinghai Tibet Plateau Herbivore Livestock Engineering Technology Center, Chengdu, China.
Front Vet Sci. 2024 Nov 4;11:1428117. doi: 10.3389/fvets.2024.1428117. eCollection 2024.
The severe conditions at high altitudes, where yaks inhabit, contribute to delayed muscular growth and compromised tenderness of their muscle tissue. Myosatellite cells are responsible for the growth and regeneration of skeletal muscle after birth and have the potential to proliferate and differentiate, its development is closely related to meat quality, and the nuclear receptor gene is involved in muscle formation and skeletal muscle regulation. Therefore, in order to understand the effect of on muscle satellite cells, we identified the mRNA expression levels of marker genes specifically expressed in muscle satellite cells at different stages to determine the type of cells isolated. Eventually, we successfully constructed a primary cell line of yak muscle satellite cells. Then we constructed overexpression vector and interference RNA, and introduced them into isolated yak skeletal muscle satellite cells. We performed qPCR, CCK8, and fluorescence-specific to detect the expression of genes or abundance of proteins as markers of cell proliferation and differentiation. Compared with those in the control group, the expression levels of proliferation marker genes , , and were significantly inhibited after overexpression, which was also supported by the CCK-8 test, whereas differentiation marker genes , , and were significantly inhibited. Fluorescence-specific staining showed that KI-67 protein abundance and the number of microfilaments both decreased, while the opposite trend was observed after interference. In conclusion, we confirmed that inhibited the proliferation and differentiation of yak skeletal muscle satellite cells, which provides a theoretical basis for further research on the effect of on improving meat quality traits and meat production performance of yaks.
牦牛栖息的高海拔恶劣环境导致其肌肉生长延迟且肌肉组织嫩度受损。肌卫星细胞负责出生后骨骼肌的生长和再生,具有增殖和分化的潜力,其发育与肉质密切相关,核受体基因参与肌肉形成和骨骼肌调节。因此,为了了解[具体基因名称]对肌肉卫星细胞的影响,我们鉴定了在不同阶段肌肉卫星细胞中特异性表达的标记基因的mRNA表达水平,以确定分离细胞的类型。最终,我们成功构建了牦牛肌肉卫星细胞原代细胞系。然后我们构建了[具体基因名称]过表达载体和干扰RNA,并将它们导入分离的牦牛骨骼肌卫星细胞中。我们进行了qPCR、CCK8和荧光特异性检测,以检测作为细胞增殖和分化标志物的基因表达或蛋白质丰度。与对照组相比,[具体基因名称]过表达后增殖标记基因[具体基因名称1]、[具体基因名称2]和[具体基因名称3]的表达水平显著受到抑制,CCK - 8试验也支持这一结果,而分化标记基因[具体基因名称4]、[具体基因名称5]和[具体基因名称6]也显著受到抑制。荧光特异性染色显示KI - 67蛋白丰度和微丝数量均减少,而[具体基因名称]干扰后观察到相反的趋势。总之,我们证实[具体基因名称]抑制了牦牛骨骼肌卫星细胞的增殖和分化,这为进一步研究[具体基因名称]对改善牦牛肉质性状和产肉性能的影响提供了理论依据。