m6A甲基化在鸡肌肉生长发育中的表达研究

doi:10.16431/j.cnki.1671-7236.2021.05.003

Abstract

Abstract: The purpose of this experiment was to study the expression of RNA m6A modification related genes ALK B homologue 5 (ALKBH5),fat mass and obesity associated protein (FTO),methyltransferase like 3 (METTLl3),methyltransferase like 14 (METTL14) and Wilms' tumor 1-associated protein (WTAP) in chicken skeletal muscle development,and analyzed its correlation with the level of skeletal muscle m6A methylation.Firstly,Real-time quantitative PCR was used to detect the mRNA expression levels of m6A methylation related genes in 12 (E12),14 (E14),16 (E16),18 (E18) embryonic and 1-day-old leg and chest muscles of Jinmao Hua chicken.And the mRNA expression levels in chicken myoblasts at 50%,100% proliferation and 1,2,3,4,5 d differentiation phases.Subsequently,the level of m6A methylation modification in the E12 and 1-day-old leg muscles and chest muscle tissues of Jinmao Hua chicken were detected by m6A methylation reagent test kit,and the correlation between the methylation level and the expression level of genes related to m6A methylation was analyzed.The results showed that the mRNA expression levels of m6A demethylation genes ALKBH5 and FTO were significantly up-regulated during skeletal muscle development (P<0.05),that wass,low expression at E12 and E14,and gradually up-regulated at E16 and E18,reaching the highest level at 1-day-old.The mRNA expression levels of the m6A methylation write genes METL14,METTL3 and WTAP gradually increased at E12,E14 and E16,and decreased at E18,and then the expression level rose back to 1-day-old.In the process of cell proliferation,the expression of ALKBH5, FTO,METL14,METL3 and WTAP genes were all up-regulated,during the process of cell differentiation,the expression levels of ALKBH5 and FTO genes were significantly up-regulated (P<0.05),reaching the highest on the fifth day of differentiation.The mRNA expression levels of METTL14,METTL3 and WTAP genes showed a downward trend on the 1st,2nd,3rd,and 4th day after cell differentiation,but increased on the 5th day after differentiation.The methylation level detection results showed that the change trend of the m6A methylation level of leg muscles and chest muscles was the same,and both decreased significantly during embryonic development (P<0.05),and reached the lowest at 1-day-old.The results of correlation analysis showed that the methylation level of chicken skeletal muscle RNA m6A was significantly negatively correlated with the mRNA expression levels of m6A demethylation modifier genes ALKBH5 and FTO (P<0.05).Based on the above experimental results,that m6A methylation modification was related to chicken skeletal muscle development,and the demethylation genes ALKBH5 and FTO might affect chicken skeletal muscle development by regulating the level of RNA m6A methylation.This study provided theoretical basis for further research on the function and molecular mechanism of m6A methylation in regulating chicken skeletal muscle growth and development.

Key words: m6A methylation; chicken; skeletal muscle development; gene expression

CLC Number:

S831.2

DING Hao, LIN Yueyue, ZHANG Tao, ZHANG Shanshan, WU Yulin, DUAN Yanjun, GONG Yongshuang, XIE Kaizhou, WANG Jinyu, DAI Guojun, ZHANG Genxi. Study on the Expression of m6A Methylation in Chicken Muscle Growth and Development[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(5): 1525-1534.

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Study on the Expression of m6A Methylation in Chicken Muscle Growth and Development

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