The Expression Characteristics Of FGF1,FGF10 And FGF21 And Their Effects On The Differentiation Of Goat Intramuscular Preadipocyte

The fibroblast growth factor(FGF)family is a kind of signaling protein with diverse functions in embryonic development,metabolism,neural function,angiogenesis and tumorigenesis.And,FGF1,FGF10 and FGF21 have been shown to play roles in white adipose tissue development,remodeling and metabolism.However,previous studies mainly focused on mice and human,and there is no report on goat.Besides,only predicted sequences of FGF1,FGF10 and FGF21 could be found in NCBI.Thus,related researches are needed to figure out whether they play roles in goat intramuscular preadipocyte differentiation and the possible mechanism.In this study,RT-PCR,real-time quantitative PCR(qPCR),cell culture,vector construction,overexpression and interference methods were used to elucidate the expression characteristics of FGF1,FGF10 and FGF21 and their effects on the differentiation of intramuscular preadipocyte in goat.The main results of this study are as follows:(1)The full-length sequences of goat FGF1,FGF10 and FGF21 were 1 165 bp,1252 bp and 666 bp,encoding 155,213 and 209 amino acids,respectively.(2)FGF1,FGF10,FGF21 and Fibroblast Growth Factor Receptors(FGFRs)gene were expressed in all examined tissues.FGF1 was highly expressed in the heart and adipose tissue.The expression level of FGF10 in lung was the highest.And FGF21 was highly expressed in liver and fat.FGFR1 and FGFR2 were mainly expressed in lung,while FGFR3 and FGFR4 were mainly expressed in the liver.(3)The recombinant adenovirus vector pAdEasy-FGF1,pAdEasy-FGF10 and pAdEasy-FGF21 were successfully constructed using pAdEasy system and successfully packaged in 293 A cells.The siRNA interference efficiency of FGF1,FGF10 and FGF21 genes were 83.9%,83.5% and 67.1%,respectively.(4)Goat intramuscular preadipocytes were obtained using collagenase II and differential adherence method.And,the expression patterns of FGF1,FGF10,FGF21 and FGFRs during goat intramuscular preadipocyte differentiation were clarified.During the differentiation of goat intramuscular preadipocyte,the expression levels of FGF1,FGF10 and FGF21 gene were extremely increased in 2 d(P<0.01)while FGFR1,FGFR2 and FGFR3 were peaked in 8 d.(5)After overexpression of FGF1,the expression level of adipocyte differentiation marker gene C/EBP? was up-regulated significantly,while PPAR?,AP2 and SREBP1 were down-regulated significantly.And FGFR1,FGFR2 and FGFR4 were up-regulated significantly.After overexpression of FGF10,the expression levels of adipocyte differentiation marker genes C/EBP?,LPL and four receptor genes were up-regulated significantly,while AP2 was down-regulated significantly.After overexpression of FGF21,the expression levels of FGFR2 and FGFR4 were up-regulated significantly,while adipocyte differentiation marker genes PPAR?,AP2 and SREBP1 were down-regulated significantly.(6)After interfering with FGF1,the expression level of FGFR2 was up-regulated and FGFR4 was down-regulated significantly,while adipocyte differentiation marker gene LPL was down-regulated significantly.After interfering with FGF10,the expression levels of adipocyte differentiation marker genes LPL,C/EBP?,AP2,SREBP1 and receptor genes FGFR1,FGFR2,FGFR3,FGFR4 were down-regulated significantly.After interfering with FGF21,the expression levels of adipocyte differentiation marker genes PPAR?,C/EBP? and SREBP1 were up-regulated while LPL was down-regulated significantly.Besides,FGFR2 and FGFR4 were down-regulated significantly.In conclusion,the above results indicate that FGF10 and FGF21 may promote and inhibit the goat intramuscular adipocytes differentiation,respectively,while the role of FGF1 remains to be studied.The results of this study provide data supports for further elucidating the molecular mechanism of FGF1,FGF10 and FGF21 gene in regulating goat intramuscular adipocytes differentiation.