Department of Animal Sciences, The Ohio State University, Wooster, OH 44691, United States of America.
Department of Animal Sciences, The Ohio State University, Wooster, OH 44691, United States of America.
Comp Biochem Physiol A Mol Integr Physiol. 2021 May;255:110917. doi: 10.1016/j.cbpa.2021.110917. Epub 2021 Feb 3.
The wooden breast (WB) myopathy is characterized by the palpation of a hard pectoralis major muscle that results in the necrosis and fibrosis of muscle fibers in fast-growing heavy weight meat-type broiler chickens. Necrosis of existing muscle fibers requires the repair and replacement of these myofibers. Satellite cells are responsible for the repair and regeneration of myofibers. To address how WB affects satellite cell function, top differentially expressed genes in unaffected and WB-affected pectoralis major muscle determined by RNA-Sequencing were studied by knocking down their expression by small interfering RNA in proliferating and differentiating commercial Ross 708 and Randombred (RBch) satellite cells. RBch satellite cells are from commercial 1995 broilers before WB appeared in broilers. Genes studied were: Nephroblastoma Overexpressed (NOV); Myosin Binding Protein-C (MYBP-C1); Cysteine-Rich Protein 3 (CSRP3); and Cartilage Oligomeric Matrix Protein (COMP). Ross 708 satellite cells had greatly reduced proliferation and differentiation compared to RBch satellite cells. MYBP-C1, CSRP3, and COMP reduced late proliferation and NOV did not affect proliferation in both lines. The timing of the knockdown differentially affected differentiation. If the expression was reduced at the beginning of proliferation, the effect on differentiation was greater than if the knockdown was at the beginning of differentiation. These data suggest, appropriate gene expression levels during proliferation greatly impact multinucleated myotube formation during differentiation. The effect of slow myofiber genes MYBP-C1 and CSRP3 on proliferation and differentiation suggests the presence of aerobic Type I satellite cells in the pectoralis major muscle which contains anaerobic Type IIb cells.
木鸡肌病的特征是可触及坚硬的胸大肌,导致快速生长的重肉质型肉鸡的肌肉纤维发生坏死和纤维化。现有肌肉纤维的坏死需要修复和替代这些肌纤维。卫星细胞负责肌纤维的修复和再生。为了研究 WB 如何影响卫星细胞功能,通过 RNA 测序确定了不受影响和受 WB 影响的胸大肌之间差异表达的前 top 基因,然后通过小干扰 RNA 敲低增殖和分化中的商业 Ross 708 和随机杂交(RBch)卫星细胞中这些基因的表达。RBch 卫星细胞来自于商业 1995 年肉鸡,此时 WB 尚未在肉鸡中出现。研究的基因有:肾母细胞瘤过表达(NOV);肌球蛋白结合蛋白 C(MYBP-C1);富含半胱氨酸的蛋白 3(CSRP3);和软骨寡聚基质蛋白(COMP)。与 RBch 卫星细胞相比,Ross 708 卫星细胞的增殖和分化能力大大降低。MYBP-C1、CSRP3 和 COMP 降低了晚期增殖,而 NOV 对两条系的增殖均无影响。敲低的时间点对分化有不同的影响。如果在增殖开始时降低表达,则对分化的影响大于在分化开始时降低表达。这些数据表明,增殖过程中适当的基因表达水平对分化过程中多核肌管的形成有很大影响。慢肌纤维基因 MYBP-C1 和 CSRP3 对增殖和分化的影响表明,胸大肌中存在有氧 Type I 卫星细胞,而该肌肉中含有无氧 Type IIb 细胞。