Proteomic Studies on the Mechanism of Myostatin Regulating Cattle Skeletal Muscle Development

Abstract

Myostatin (MSTN) is an important negative regulator of muscle growth and development. In this study, we performed comparatively the proteomics analyses of gluteus tissues from MSTN+/− Mongolian cattle (MG.MSTN+/−) and wild type Mongolian cattle (MG.WT) using a shotgun-based tandem mass tag (TMT) 6-plex labeling method to investigate the regulation mechanism of MSTN on the growth and development of bovine skeletal muscle. A total of 1,950 proteins were identified in MG.MSTN+/− and MG.WT. Compared with MG.WT cattle, a total of 320 differentially expressed proteins were identified in MG.MSTN cattle, including 245 up-regulated differentially expressed proteins and 75 down-regulated differentially expressed proteins. Bioinformatics analysis showed that knockdown of the MSTN gene increased the expression of extracellular matrix and ribosome-related proteins, induced activation of focal adhesion, PI3K-AKT, and Ribosomal pathways. The results of proteomic analysis were verified by muscle tissue Western blot test and in vitro MSTN gene knockdown test, and it was found that knockdown MSTN gene expression could promote the proliferation and myogenic differentiation of bovine skeletal muscle satellite cells (BSMSCs). At the same time, Co-Immunoprecipitation (CO-IP) assay showed that MSTN gene interacted with extracellular matrix related protein type I collagen α 1 (COL1A1), and knocking down the expression of COL1A1 could inhibit the activity of adhesion, PI3K-AKT and ribosome pathway, thus inhibit BSMSCs proliferation. These results suggest that the MSTN gene regulates focal adhesion, PI3K-AKT, and Ribosomal pathway through the COL1A1 gene. In general, this study provides new insights into the regulatory mechanism of MSTN involved in muscle growth and development.

Multi-Omics Approach Reveals Dysregulation of Protein Phosphorylation Correlated with Lipid Metabolism in Mouse Non-Alcoholic Fatty Liver

“…We then performed a Student’s t -test to unveil proteins that were significantly regulated after DIO. A volcano plot representation of our filtered data shows that the distribution of upregulated and downregulated proteins was remarkably symmetrical, with 312 proteins above the q < 0.05 and 0.38 |log 2 (fold-change)| cutoffs [ 21 , 22 , 23 , 24 , 25 ], set as the minimum to consider proteins as “significantly regulated” ( Figure 3 B). Specifically, 141 proteins were downregulated and 171 were upregulated.…”

Growth Traits and Sperm Proteomics Analyses of Myostatin Gene-Edited Chinese Yellow Cattle

“…Therefore, the sperm motility of MT bulls might be enhanced, and the flagellum fiber of sperm might be increased. Our previous reports have shown that the glycolysis, glycogen metabolism, and fatty acid β-oxidation was significantly changed in MT cattle [ 37 , 46 , 47 , 48 ]. Meanwhile, various effects of MSTN gene mutation on cattle and the related molecule mechanism have been investigated, including bile acid metabolism [ 19 ], enhanced glycolysis in the heart [ 37 ], and microbial community of stomach [ 36 ].…”

Transcriptomic Analysis Reveals mRNA and Alternative Splicing Events in Ovine Skeletal Muscle Satellite Cells during Proliferation and Differentiation

“…For instance, ACTC1 (actin alpha cardiac muscle 1) is involved in the differentiation of myoblasts, and ACTC1 deficiency led to severe structural and functional perturbations in the heart [ 44 , 45 ]. The knockdown of COL1A1 inhibits the proliferation of bovine skeletal muscle satellite cells [ 46 ]. Therefore, these DEGs are indispensable to the proliferation and differentiation of SMSCs and to muscle development.…”