Transcriptome sequencing to explore the effect of miR-214 on chicken primary myoblasts

Abstract

MicroRNAs are involved in a series of biological processes, such as proliferation, differentiation and apoptosis of primary myoblasts. The research group found that miR-214 is highly expressed in chicken primary myoblasts (CPMs), so we used miR-214 as a starting point to explore the biological function of miR-214 in skeletal muscle growth and development. In this experiment, CPMs were cultured in vitro; miR-214 was overexpressed in CPMs; and cell samples were collected for subsequent transcriptome sequencing (RNA-seq). After miR-214 overexpression, we identified 97 differentially expressed genes (DEGs), of which 21 DEGs were up-regulated and 76 DEGs were down-regulated. After bioinformatics analysis, these DEGs were found to be significantly enriched in myofibrils, muscle system processes, myofibril assembly and other biological processes related to muscle development. The significantly enriched KEGGs include focal adhesion and type II diabetes mellitus. The protein network of DEGs was drawn by STRING and Cytoscape software, and 5 DEGs were randomly selected to verify the sequencing results by real-time fluorescence quantification. CAV3 is not only an important node protein in the protein network but also a member of the focal adhesion signaling pathway. It is speculated that miR-214 may regulate muscle development through the focal adhesion signaling pathway.

Keywords:

• Chicken
• myoblast
• miR-214
• RNA-seq
• up-regulated
• real-time PCR

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

The data presented in this study are openly available in the SRA database; the accession number is PRJNA773374.

Additional information

Funding

This work was jointly supported by the Natural Science Foundation of Jiangsu Province (BK20181453), the New Agricultural Breeds Creation Project in Jiangsu Province (PZCZ201730), the Special Funds Project for Transforming Scientific and Technological Achievements in Jiangsu Province (BA2018099), the China Agriculture Research System (CARS-41) and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).