Geng Wanzhuo, Guo Yulong, Chen Botong, Cheng Xi, Li Shuohan, Challioui Mohammed Kamal, Tian Weihua, Li Hong, Zhang Yanhua, Li Zhuanjian, Jiang Ruirui, Tian Yadong, Kang Xiangtao, Liu Xiaojun
College of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China.
College of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China; Animal Production and Biotechnology Department, Institut Agronomique et Vétérinaire Hassan II, Rabat P.O. Box 6202, Rabat, Morocco.
Poult Sci. 2024 Dec;103(12):104258. doi: 10.1016/j.psj.2024.104258. Epub 2024 Sep 2.
Though it is well known that insulin-like growth factor (IGF) binding protein 7 (IGFBP7) plays an important role in myogenesis and adipogenesis in mammals, its impact on the proliferation, differentiation, and lipid deposition in chicken primary myoblasts (CPM) and intramuscular preadipocytes remains unexplored. In the present study, we firstly examined the correlation between SNPs within the genomic sequence of the IGFBP7 gene and carcass and blood chemical traits in a F2 resource population by genetic association analysis, and found that a significant correlation between the SNP (4_49499525) located in the intron region of IGFBP7 and serum high-density lipoproteins (HDL). We then examined the expression patterns of IGFBP7 across different stages of proliferation and differentiation in CPMs and intramuscular preadipocytes via qPCR, and explored the biological functions of IGFBP7 through gain- and loss-of-function experiments and a range of techniques including qPCR, CCK-8, EdU, flow cytometry, Western blot, immunofluorescence, and Oil Red O staining to detect the proliferation, differentiation, and lipid deposition in CPMs and intramuscular preadipocytes. We ascertained that the expression levels of the IGFBP7 gene increased as cell differentiation progresses in CPMs and intramuscular preadipocytes, and that IGFBP7 promotes the proliferation and differentiation of these cells, as well as facilitates intracellular lipid deposition. Furthermore, we investigated the regulatory mechanism of IGFBP7 expression by using co-transfection strategy and dual-luciferase reporter assay, and discovered that the myogenic transcription factors (MRF), myoblast determination factor (MyoD) and myogenin (MyoG), along with the adipocyte-specific transcription factor (TF) CCAAT/enhancer-binding protein α (C/EBPα), can bind to the core transcription activation region of the IGFBP7 promoter located 500 bp upstream from the transcription start site, thereby promoting IGFBP7 transcription and expression. Taken together, our study underscores the role of IGFBP7 as a positive regulator for myogenesis and adipogenesis, while also elucidating the functional and transcriptional regulatory mechanisms of IGFBP7 in chicken skeletal muscle development and intramuscular adipogenesis.
虽然众所周知胰岛素样生长因子(IGF)结合蛋白7(IGFBP7)在哺乳动物的肌生成和脂肪生成中起重要作用,但其对鸡原代成肌细胞(CPM)和肌内前脂肪细胞的增殖、分化及脂质沉积的影响仍未得到探索。在本研究中,我们首先通过遗传关联分析研究了IGFBP7基因基因组序列内的单核苷酸多态性(SNP)与F2资源群体的胴体和血液化学性状之间的相关性,发现位于IGFBP7内含子区域的SNP(4_49499525)与血清高密度脂蛋白(HDL)之间存在显著相关性。然后,我们通过qPCR检测了IGFBP7在CPM和肌内前脂肪细胞增殖及分化不同阶段的表达模式,并通过功能获得和功能丧失实验以及包括qPCR、CCK - 8、EdU、流式细胞术、蛋白质免疫印迹、免疫荧光和油红O染色等一系列技术,探索了IGFBP7在CPM和肌内前脂肪细胞中的增殖、分化及脂质沉积方面的生物学功能。我们确定,在CPM和肌内前脂肪细胞中,随着细胞分化的进行,IGFBP7基因的表达水平升高,并且IGFBP7促进这些细胞的增殖和分化,同时促进细胞内脂质沉积。此外,我们通过共转染策略和双荧光素酶报告基因检测研究了IGFBP7表达的调控机制,发现成肌转录因子(MRF)、成肌细胞决定因子(MyoD)和肌细胞生成素(MyoG),以及脂肪细胞特异性转录因子(TF)CCAAT/增强子结合蛋白α(C/EBPα),可以结合到转录起始位点上游500 bp处的IGFBP7启动子的核心转录激活区域,从而促进IGFBP7的转录和表达。综上所述,我们的研究强调了IGFBP7作为肌生成和脂肪生成的正调节因子的作用,同时也阐明了IGFBP7在鸡骨骼肌发育和肌内脂肪生成中的功能及转录调控机制。