Multiple Spliced Transcripts And Differential Expression Pattern Of The Myocyte Enhancer Factor 2c(MEF2C) Gene In Goat(Capra Hircus)

Myocyte Enhancer Factor 2C(MEF2C)is a member of the Myocyte Enhancer Factor 2(MEF2)family.MEF2C is a important transcription factors to participate in a variety of biologicals.mRNA precursors of MEF2C gene produce multiple mature mRNA alternative splicing through different splicing,to translate into different proteins.Therefore,MEF2C gene may encode a variety of proteins.In people and mice,several alternative spliced transcripts of MEF2C have been identified,respectively.In the goats,MEF2C gene may exist 10 alternative splicing by transcriptome sequencing,but identified and validated through the experiment of goats MEF2C alternative splicing has not been reported.Here,we selected Nanjiang Brown goats as experimental material.RT-PCR,molecular cloning,qPCR,TNT system,western blotting and goats SMSCs in vitro culture techniques were used to clone goat MEF2C gene,identify goat MEF2C gene transcription,and further to determine the level of mRNA and protein expression pattern of different MEF2C transcripts in different tissues.The results are as follows.1.We characterized and identified four MEF2C spliced variants in different goat tissues and designated as MEF2C1,MEF2C2,MEF2C3 and MEF2C4.The cDNA sequences of the four goat MEF2C transcripts were deposited in GenBank as KY457285-KY457288.Compared with the whole caprine genome sequences,MEF2C1 lacked of the Exon8,Exonll and Exon12.MEF2C2 lacked of the Exon7 to Exon.12 MEF2C3 lacked of the Exon8 and Exon 12.MEF2C3 lacked of the Exon7,Exon8 and Exon 12.2.The function domain of the four MEF2C transcripts showed that four transcripts are included by MADS function domain,but MEF2C2 and MEF2C4 missing HJURP C function domain.MEF2C1 has a low matching degree of functional region,but MEF2C2,MEF2C3 and MEF2C4 has two atypical low matching degree of functional region.3.qPCR analyses revealed that MEF2C1 and MEF2C2 transcripts have higher levels of expression in longissimus dorsi(P<0.01),and MEF2C3 has higher levels of expression in the semitendinosus and psoas major(P<0.01),but all the MEF2C transcripts were in the liver and kidney have low levels of expression.4.qPCR detection MEF2C four transcripts in five different times of muscle tissues(3 d,30 d,60 d,90 d and 120 d),four transcriptions displayed along with the increase of age after the first increase to reduce,30 d reached the highest(P<0.01),the expression of 120 d relative to the minimum.5.MEF2C1,MEF2C3 and MEF2C4 have higher levels of expression in differentiation of 1 d(D1)(P<0.01),and MEF2C2 transcript high levels of expression in differentiation 3 d(D3)(P<0.01),but the expression of four transcriptions in the proliferation period are low,relatively.6.Western blotting revealed that abundant MEF2C proteins were observed in the higher molecular weight band(50 kDa)obtained from the heart,liver and psoas major,in the brain and longissimus dorsi tissue expression in the weak.In the lower molecular weight band(25 kDa),MEF2C can be detected in psoas major,semimembranosus and longissimus dorsi tissues,but the profile in kidney is low.Moreover MEF2C protein can’t be detected in liver.7.Overexpression of MEF2C2 significantly reduce the expression of MyoG(P<0.05),and it indicated that MEF2C2 has inhibitory effect on cell differentiation.