College of Animal Science and Technology, Henan Agricultural University, Zhengzhou, 450046, China.
Henan Key Laboratory for Innovation and Utilization of Chicken Germplasm Resources, Zhengzhou, 450046, China.
Commun Biol. 2024 May 2;7(1):518. doi: 10.1038/s42003-024-06226-1.
Myoblast proliferation and differentiation are essential for skeletal muscle development. In this study, we generated the expression profiles of mRNAs, long noncoding RNAs (lncRNAs), and microRNAs (miRNAs) in different developmental stages of chicken primary myoblasts (CPMs) using RNA sequencing (RNA-seq) technology. The dual luciferase reporter system was performed using chicken embryonic fibroblast cells (DF-1), and functional studies quantitative real-time polymerase chain reaction (qPCR), cell counting kit-8 (CCK-8), 5-Ethynyl-2'-deoxyuridine (EdU), flow cytometry cycle, RNA fluorescence in situ hybridization (RNA-FISH), immunofluorescence, and western blotting assay. Our research demonstrated that miR-301a-5p had a targeted binding ability to lncMDP1 and ChaC glutathione-specific gamma-glutamylcyclotransferase 1 (CHAC1). The results revealed that lncMDP1 regulated the proliferation and differentiation of myoblasts via regulating the miR-301a-5p/CHAC1 axis, and CHAC1 promotes muscle regeneration. This study fulfilled the molecular regulatory network of skeletal muscle development and providing an important theoretical reference for the future improvement of chicken meat performance and meat quality.
成肌细胞的增殖和分化对于骨骼肌的发育至关重要。在这项研究中,我们使用 RNA 测序 (RNA-seq) 技术生成了鸡原代成肌细胞 (CPM) 不同发育阶段的 mRNA、长非编码 RNA (lncRNA) 和 microRNA (miRNA) 的表达谱。使用鸡胚成纤维细胞 (DF-1) 进行双荧光素酶报告系统检测,通过定量实时聚合酶链反应 (qPCR)、细胞计数试剂盒-8 (CCK-8)、5-乙炔基-2'-脱氧尿苷 (EdU)、流式细胞术周期、RNA 荧光原位杂交 (RNA-FISH)、免疫荧光和 Western blot 检测进行功能研究。我们的研究表明,miR-301a-5p 与 lncMDP1 和 ChaC 谷胱甘肽特异性γ-谷氨酰环转移酶 1 (CHAC1) 具有靶向结合能力。结果表明,lncMDP1 通过调节 miR-301a-5p/CHAC1 轴来调节成肌细胞的增殖和分化,而 CHAC1 促进肌肉再生。本研究揭示了骨骼肌发育的分子调控网络,为未来提高鸡肉性能和改善肉质提供了重要的理论参考。