Summary: | Muscle development is a complex biological process involving an intricate network of multiple factor interactions. Through the analysis of transcriptome data and molecular biology confirmation, this study aims to reveal the molecular mechanism underlying sheep embryonic skeletal muscle development. The RNA sequencing of embryos was conducted, and microRNA (miRNA)-mediated competitive endogenous RNA (ceRNA) networks were constructed. qRT-PCR, siRNA knockdown, CCK-8 assay, scratch assay, and dual luciferase assay were used to carry out gene function identification. Through the analysis of the ceRNA networks, three miRNAs (miR-493-3p, miR-3959-3p, and miR-410-5p) and three genes (<i>TEAD1</i>, <i>ZBTB34,</i> and <i>POGLUT1</i>) were identified. The qRT-PCR of the DE-miRNAs and genes in the muscle tissues of sheep showed that the expression levels of the <i>TEAD1</i> gene and miR-410-5p were correlated with the growth rate. The knockdown of the <i>TEAD1</i> gene by siRNA could significantly inhibit the proliferation of sheep primary embryonic myoblasts, and the expression levels of <i>SLC1A5, FoxO3, MyoD</i>, and <i>Pax7</i> were significantly downregulated. The targeting relationship between miR-410-5p and the <i>TEAD1</i> gene was validated by a dual luciferase assay, and miR-410-5p can significantly downregulate the expression of <i>TEAD1</i> in sheep primary embryonic myoblasts. We proved the regulatory relationship between miR-410-5p and the <i>TEAD1</i> gene, which was related to the proliferation of sheep embryonic myoblasts. The results provide a reference and molecular basis for understanding the molecular mechanism of embryonic muscle development.
|