Myostatin通过激活TRPC4/Ca2+/Calcineurin/NFATc3通路促进牛肌卫星细胞增殖

doi:10.1016/j.jia.2024.04.020

Journal of Integrative Agriculture ›› 2026, Vol. 25 ›› Issue (3): 1125-1136.DOI: 10.1016/j.jia.2024.04.020

Myostatin通过激活TRPC4/Ca²⁺/Calcineurin/NFATc3通路促进牛肌卫星细胞增殖

收稿日期:2023-10-31 修回日期:2024-05-16 接受日期:2024-03-13 出版日期:2026-03-20 发布日期:2026-02-06

Myostatin promotes proliferation of bovine muscle satellite cells through activating TRPC4/Ca²⁺/calcineurin/NFATc3 pathway

Yajie Gao^{1, 2*}, Song Wang^{1, 3*}, Anqi Di¹, Chao Hai¹, Di Wu¹, Zhenting Hao¹, Lige Bu¹, Xuefei Liu¹, Chunling Bai¹, Guanghua Su¹, Lishuang Song¹, Zhuying Wei¹, Zhonghua Liu³, Lei Yang^1#, Guangpeng Li^1#

¹State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, College of Life Science, Inner Mongolia University, Hohhot 010070, China
²The Central Lab, The First Affiliated Hospital of Baotou Medical College, Baotou 014010, China
³College of Life Science, Northeast Agricultural University, Harbin 150030, China

Received:2023-10-31 Revised:2024-05-16 Accepted:2024-03-13 Online:2026-03-20 Published:2026-02-06
About author:Yajie Gao, E-mail: gaoyajie1997@163.com; Song Wang, E-mail: wangsong199852@163.com; #Correspondence Lei Yang, Tel: +86-471-5298583, E-mail: leiyang@imu.edu.cn; Guangpeng Li, Tel: +86-471-5298583, E-mail: gpengli@imu.edu.cn * These authors contributed equally to this study.
Supported by:
This study was supported by the National Natural Science Foundation of China (32360837 and 32341052), the STI2030 Major Projects, China (2023ZW0404803), the Inner Mongolia Open Competition Projects, China (2022JBGS0025), the Inner Mongolia Science and Technology Leading Team, China (2022LJRC0006), the Inner Mongolia Science and Technology Major Projects, China (2021ZD0009, 2021ZD0008, 2022ZD0008, 2023KJHZ0028), the Inner Mongolia Young Talents Projects, China (NJYT23138), the Inner Mongolia Natural Science Foundation, China (2023MS03004), the Central Government Guides Development, China (2022ZY0212), the National Agricultural Science and Technology Project of China (NK2022130203), the Collaborative Innovation among Universities in Hohhot, China (XTCX2023-06), and the Ministry of Education Engineering Centre Project, China (JYBGCSYS2022).

摘要/Abstract

摘要：

肌生成抑制素（myostatin，MSTN）主要在骨骼肌中表达，MSTN基因的缺失会促进肌细胞增殖。Ca²⁺是生物体中广泛存在的第二信使，对于肌细胞的增殖和肌肉生长至关重要。然而，MSTN基因与Ca²⁺之间的联系目前仍不清楚。因此，本研究以MSTN基因编辑牛（MSTN^-/-）作为研究对象，从Ca²⁺通道及其调控途径角度探讨MSTN突变促进肌卫星细胞增殖的分子机制。与野生型（WT）相比，MSTN突变促进牛肌卫星细胞增殖。通过电感耦合等离子体—质谱法(Inductively coupled plasma-Mass Spectrometry，ICP-MS）发现，MSTN^-/-牛的肌肉组织和肌卫星细胞中Ca²⁺含量显著升高。同时，转录组测序发现MSTN^-/-牛的TRPC4表达水平显著增加。这些结果表明，MSTN突变激活Ca²⁺通道TRPC4促进细胞增殖。通过使用TRPC4的特异性抑制剂ML204处理MSTN^-/-肌卫星细胞，发现Ca²⁺含量显著降低至WT水平。Calreticulin的Western blot和特异性染色发现，ML204处理后MSTN^-/-肌卫星细胞中Calcineutrin的表达量和NFATc3入核量显著降低，细胞增殖能力受到抑制。综上所述，MSTN突变激活钙离子通道TRPC4，引起胞质内Ca²⁺含量升高，激活Calcineurin活性，致使NFATc3去磷酸化入核，激活增殖相关基因从而促进肌卫星细胞增殖，使MSTN^-/-牛出现“双肌”表型。本研究揭示了MSTN突变激活TRPC4/Ca²⁺/Calcineurin/NFATc3通路促进肌肉发育的分子机制，为进一步探究MSTN对牛肌肉发育的调控机制提供参考。

Abstract:

Myostatin (MSTN) is principally expressed in skeletal muscle and negatively regulates muscle growth and development. MSTN mutation can induce muscle overgrowth in cattle by activating cell proliferation, presenting a “double-muscle” phenotype. However, the specific regulatory mechanism is still unclear. Here, we found that Ca²⁺ content in muscle tissue and muscle satellite cells of MSTN mutated (MSTN^–/–) cattle were significantly increased compared to wild-type (WT). Furthermore, transcriptome analysis of muscle satellite cells revealed that TRPC4 was significantly increased in MSTN^–/–cattle. And the expression of TRPC4 in muscle tissue of MSTN^–/– cattle was detected by RT-qPCR and Western blot, which was significantly higher than that of WT. These results suggested that MSTN mutation promoted muscle satellite cells proliferation through activation of TRPC4 channel. To further verify, ML204, a specific inhibitor of TRPC4, was used to treat MSTN^–/– muscle satellite cells. We found that cell proliferation was inhibited, calcineurin expression was downregulated, and the entry of NFATc3 into nuclei was reduced, which was similar to WT group. Thus, MSTN mutation leads to the activation of TRPC4 channel, which increases intracellular Ca²⁺content, further activates calcineurin/NFATc3 pathway, and ultimately promotes the proliferation of muscle satellite cells.

Key words: myostatin , cell proliferation , Ca²⁺ , TRPC4 , NFATc3

Yajie Gao, Song Wang, Anqi Di, Chao Hai, Di Wu, Zhenting Hao, Lige Bu, Xuefei Liu, Chunling Bai, Guanghua Su, Lishuang Song, Zhuying Wei, Zhonghua Liu, Lei Yang, Guangpeng Li. Myostatin通过激活TRPC4/Ca²⁺/Calcineurin/NFATc3通路促进牛肌卫星细胞增殖[J]. Journal of Integrative Agriculture, 2026, 25(3): 1125-1136.

Yajie Gao, Song Wang, Anqi Di, Chao Hai, Di Wu, Zhenting Hao, Lige Bu, Xuefei Liu, Chunling Bai, Guanghua Su, Lishuang Song, Zhuying Wei, Zhonghua Liu, Lei Yang, Guangpeng Li. Myostatin promotes proliferation of bovine muscle satellite cells through activating TRPC4/Ca²⁺/calcineurin/NFATc3 pathway[J]. Journal of Integrative Agriculture, 2026, 25(3): 1125-1136.

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Myostatin通过激活TRPC4/Ca²⁺/Calcineurin/NFATc3通路促进牛肌卫星细胞增殖

Myostatin promotes proliferation of bovine muscle satellite cells through activating TRPC4/Ca²⁺/calcineurin/NFATc3 pathway

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