The MyoD1 Promoted Muscle Differentiation and Generation by Activating CCND2 in Guanling Cattle

Di Zhou; Yan Wang; Rong Yang; Fu Wang; Zhonghai Zhao; Xin Wang; Lingling Xie; Xingzhou Tian; Guoze Wang; Bo Li; Yu Gong

doi:10.3390/ani12192571

The MyoD1 Promoted Muscle Differentiation and Generation by Activating CCND2 in Guanling Cattle

Animals (Basel). 2022 Sep 26;12(19):2571. doi: 10.3390/ani12192571.

Authors

Di Zhou¹, Yan Wang¹, Rong Yang¹, Fu Wang¹, Zhonghai Zhao¹, Xin Wang¹, Lingling Xie¹, Xingzhou Tian², Guoze Wang², Bo Li³, Yu Gong³

Affiliations

¹ Guizhou Testing Center for Livestock and Poultry Germplasm, Guiyang 550018, China.
² Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountainous Region, Ministry of Education, Guizhou University, Guiyang 550025, China.
³ Guizhou Livestock and Poultry Genetic Resources Management Station, Guiyang 550001, China.

Abstract

The purpose of this study was to analyze the transcriptome of MyoD1 gene knockout MDBK cells (bovine kidney cells) using high-throughput sequencing. For the first time, CRISPR/CAS9 technology was used to construct a MyoD1 knockout in MDBK cells and transcriptome sequence analysis was used to examine MyoD1-related target gene expression. Transcriptome sequencing indicated the presence of 723 differentially expressed genes (DEGs) by comparing wild type and MyoD1 knockout MDBK cells and included 178 upregulated and 72 downregulated genes. The DEGs are mainly enriched in Pl-3-kinase and AKT, p53 signaling pathways. Quantitative RT-PCR confirmed that PDE1B, ADAMTS1, DPT, and CCND2 were highly expressed in the leg muscle, longissimus dorsi, and shoulder of Guanling cattle, and CCND2 was inhibited after MyoD1 knockout, suggesting it may be a key downstream gene of MyoD1 and associated with muscle formation and differentiation in Guanling cattle. This provides experimental data for subsequent studies on the regulatory mechanisms of muscle differentiation in Guanling cattle.

Keywords: CRISPR/CAS9; Guanling cattle; MyoD1; transcriptome.

Grants and funding

The project was supported by the Central Guidance to Local Funds Grant (QKZY-2018-4015); the study on comprehensive utilization and anti-oxidation technology of main crop straw and forage resources in Guizhou [2019]2367; the Guizhou Provincial Academy of Agricultural Sciences-level Innovation Fund Project (Qiankehe Platform Talents [2018]5768-08); and the Guizhou Agricultural Animal and Plant Breeding Project (Guizhou Agricultural Breeding Zhuanzi [2018] 016).