MEF2A Promoter Methylation Negatively Regulates mRNA Transcription in Guanling Cattle

doi:10.7668/hbnxb.20195667

Abstract

Abstract:

Muscle formation depends on the interplay of various cellular and extracellular signals and factors.This study investigates the regulatory relationship between Myocyte enhancer factor 2A(MEF2A)expression levels and promoter methylation at both tissue and cellular levels,to provide theoretical references for the genetic development of Guanling cattle.Guanling cattle were used as experimental subjects.Initially,the expression levels of the MEF2A in Guanling cattle tissues were analyzed in conjunction with their promoter methylation status.Subsequently,overexpression and interference vectors of the MEF2A gene were successfully transfected into primary myoblasts of Guanling cattle,and the effects of MEF2A expression level changes on promoter methylation levels were analyzed.The results showed that the MEF2A expression levels in various tissues of young Guanling cattle were higher than those in adult cattle,while the methylation rate of MEF2A in young cattle tissues was lower than that in adult cattle,with DNMT1 expression trends consistent with this.Additionally,increased MEF2A expression led to a decrease in its methylation rate,whereas decreased MEF2A expression resulted in an increase in its methylation rate.This study confirms from both tissue and cellular perspectives that the expression level of the MEF2A gene in Guanling cattle is negatively correlated with its methylation rate,providing a theoretical basis for genetic marker-assisted genetic improvement of cattle.

Key words: Cuanling cattle, MEF2A, Promoter, DNA methylation, Myoblasts

CLC Number:

S823

SUN Jinkui, XU Houqiang. MEF2A Promoter Methylation Negatively Regulates mRNA Transcription in Guanling Cattle[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(4): 203-209. doi: 10.7668/hbnxb.20195667.

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Figures/Tables 6

Tab.1 Primers sequence information

名称 Name	引物序列(5'-3') Primer sequence(5'-3')	长度/bp Length
MEF2A	F: AATGAACCTCACGAAAGCAGAAC	106
	R:TTAGCACATAGGAAGTATCAGGGTC
DNMT1	F: GCACAGTCAAGGCAGAGAA	132
	R: CCAGCCTTCTCCATGGTAGT
GAPDH	F: TTGTGATGGGCGTGAACC	169
	R: GTCTTCTGGGTGGCAGTGAT
CPG-1	F: ATTATTATTAGTAGTAAAGATTAGTAGG	520
	R: ACCTCCCAAAACAACC
CPG-2	F: GGTTGTTTTGGGAGGT	110
	R: CTCCTAAAAAAAATATTAATCA
CPG-3	F: GTGATTAATATTTTTTTTAGGAGAATA	246
	R: TCTTCTTAATAAAACCCAAAACACTTAA

Fig.1 Expression of MEF2A CpG island(A)and its expression in Guanling cattle tissues(B) Different lowercase letters represent significant differences(P<0.05);different capital letters represent extremely significant(P<0.01).The same as Fig.2,4.

Tab.2 RNA quality information

组织 Tissue	犊牛Calves		成年牛Adult
组织 Tissue	浓度/ (ng/μL) Concentration	OD_260/280	浓度/ (ng/μL) Concentration	OD_260/280
心脏Heart	1 082.3	1.86	1 124.7	1.85
肝脏Liver	985.6	1.91	1 356.9	1.92
肺脏Lung	1 428.5	1.93	1 003.8	1.86
肾脏Kidney	1 126.8	1.83	895.4	1.91
背最长肌	879.2	1.87	1 156.7	1.82
Dorsal muscle
脂肪Fat	685.1	1.81	752.6	1.81

Fig.2 Effects of MEF2A on heart and adipose methylation rate(A)and DNMT1 expression(B)in cattle

Fig.3 Immunofluorescence identification(200×)

Fig.4 Effects of recombinant vectors on MEF2A promoter methylation rate and DNMT1 expression A. MEF2A overexpression vector efficiency;B.The effect of overexpression of MEF2A on methylation rate;C.The effect of overexpression of MEF2A on DNMT1;D.MEF2A interference carrier efficiency;E.The effect of inhibition of MEF2A on methylation rate;F.The effect of inhibit in of MEF2A on DNMT1.oe-NC.Overexpression negative control vector;oe-MEF2A.Overexpression vector;sh-NC.Interference negative control vector;sh-MEF2A.Interference vector.

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