The identification and validation of target genes of IGFBP3 protein in sheep skeletal muscle cells

Abstract

This study aimed to identify the target genes of IGFBP3（insulin growth factor binding protein）protein and to investigate its target genes effects on the proliferation and differentiation of Hu sheep skeletal muscle cells. IGFBP3 was an RNA-binding protein that regulates mRNA stability. Previous studies have reported that IGFBP3 promotes the proliferation of Hu sheep skeletal muscle cells and inhibits differentiation, but the downstream genes that bind to it have not been reported yet. We predicted the target genes of IGFBP3 through RNAct and sequencing data, and verified by qPCR and RIP（RNA Immunoprecipitation）experiments, and demonstrated GNAI2（G protein subunit alpha i2）as one of the target gene of IGFBP3. After interference with siRNA, we carried out qPCR, CCK8, EdU, and immunofluorescence experiments, and found that GNAI2 can promote the proliferation and inhibit differentiation of Hu sheep skeletal muscle cells. This study revealed the effects of GNAI2 and provided one of the regulatory mechanisms of IGFBP3 protein underlying sheep muscle development.

Keywords:

• IGFBP3
• GNAI2
• proliferation
• differentiation
• skeletal muscle cell
• Hu sheep

Author Contributions

Sun Wei and CAO Xiukai designed this study; WANG Shan performed the experiments and wrote the manuscript; GU Yifei, Ge Ling, He Mingliang and Zhang Weibo contributed to materials and data collection in this study; Sun Wei, CAO Xiukai, Tesfaye Getachew, Joram M. Mwacharo and Aynalem Haile revised the final manuscript. All authors approved the final manuscript.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This work was supported by the National Natural Science Foundation of China (31872333, 32172689), National Natural Science Foundation of China-CGIAR (32061143036), Major New Varieties of Agricultural Projects in Jiangsu Province (PZCZ201739), Natural Science Foundation of the Jiangsu Higher Education Institutions of China [22KJA230001], Major Project of Natural Science Foundation of Xinjiang Uyghur Autonomous Region [2022D01D47], Jiangsu 333 Distinguished Talents Project Foundation [(2022) 2-323], Distinguished Talents Project Foundation of Yangzhou University, High-end Foreign Expert Introduction Project (G2022014148L), Jiangsu Natural Science Foundation for Colleges and Universities [21KJB230005], and Jiangsu Postdoctoral Science Foundation [2021K608C].