The Regulatory Effect Of FGF10 Gene On The Differentiation Of Bovine Preadipocytes

Beef is rich in nutrients,low in cholesterol,high in protein,and rich in vitamins,calcium and other nutrients.Due to the improvement of people’s living standards,the demand for beef has increased.Tenderness,flavor and marbling of beef are one of the main quality indicators of its beef.Beef tenderness and marbling are closely related to beef intramuscular fat content.In mammals,adipose tissue is mainly stored in subcutaneous,intrafat,intramuscular and intermuscular.In the process of meat production,the amount of intramuscular fat is the main factor determining the quality of meat.At the cellular level,the amount of fat in an animal is closely related to the number and size of fat cells.In addition,the regulation of differentiation-related genes is required in the process of adipocyte differentiation.At present,it is a very important research direction to study the key genes that affect the differentiation of adipocytes.Fibroblast growth factor 10(FGF10)belongs to the FGF7 subfamily among the 6FGFs subfamily.It is the only FGF specifically expressed in white adipose tissue and plays an important role in the differentiation of adipose tissue..Studies have shown that in the FGF family,the FGF10 gene is specifically expressed in white adipose tissue and promotes fat formation.Studies have reported that FGF10 gene is abundantly expressed in the subcutaneous fat,abdominal wall and visceral fat adipose tissue of extremely obese patients.However,studies on the regulation of FGF10 on preadipocyte differentiation are rarely reported.Therefore,this study took the new line of Qinchuan beef cattle selected by Northwest Agriculture and Forestry University as the research object,and studied the regulatory mechanism of FGF10 in the differentiation of preadipocytes,aiming to provide the genetic improvement and molecular breeding for Qinchuan beef cattle.Theoretical basis.The main findings are as follows:1.In this study,the expression level of FGF10 gene in different bovine tissues was detected at the m RNA expression level.The results showed that the expression of FGF10 gene in bovine ostium tissue was significantly higher than that in other tissues,and the expression level in subcutaneous fat and internal fat tissue was also higher.high,significantly higher than other tissues,this result indicates that FGF10 gene may have an important role in adipogenesis,adipose differentiation and metabolism.In this study,the relative expression levels of adipocyte differentiation marker genes C/EBPa,PPARy,FABP4,FAS and LPL were detected by q PCR after infecting cells with Ad-FGF10 and transfecting siFGF10 respectively.It was found that after overexpression of FGF10 gene,the expression levels of C/EBPa,PPARy,FABP4 and LPL were significantly down-regulated compared with the control group on 4day of induced differentiation(P?0.01,P?0.05).PPARy,FABP4 and LPL were significantly increased at 6d of induced differentiation(P?0.05).From the expression changes of C/EBPa,PPARy,FABP4 and LPL,it can be seen that FGF10 can inhibit the differentiation of bovine adipocytes.The expression levels of PPARγ and FABP4 were detected at the protein level,and it was found that the protein expression levels of FGF10 overexpressed were significantly decreased compared with the control group,while the expression levels of PPARγ and FABP4 were significantly increased after interference compared with the control group,indicating that FGF10 Targeted inhibition of PPARγ and FABP4 genes.Oil red O staining and triglyceride content determination showed that overexpression of FGF10 gene inhibited the accumulation of triglycerides in bovine adipocytes,while interference with FGF10 gene promoted the accumulation of triglycerides in bovine adipocytes.2.The related candidate genes of bovine preadipocytes overexpressing adenovirus Ad-FGF10(experimental group)and Ad-NC induced differentiation were analyzed by transcriptome sequencing on the 4th day.A total of 1774 differential genes were detected,of which 157 genes were significantly up-regulated and 1617 genes were significantly down-regulated.In this study,through the significant enrichment analysis of GO function and the significant enrichment analysis of KEGG signaling pathway,differential genes are involved in regulating a wide range of biological processes(such as metabolism,biological process regulation,cell proliferation),and are enriched in lipid differentiation-related genes.PPAR signaling pathway,regulation of lipolysis in adipocytes,fatty acid degradation and other pathways.3.Three SNP sites were identified in the promoter region of FGF10 gene,namely g.78G>A,g.116C>T and g.201A>T.In 200 Qinchuan cattle,3 SNPs were found to have 3different genotypes.Among them,SNP1 had GG,GA and AA genotypes;SNP2 had CC,CT and TT genotypes,and SNP3 had AA,AT and TT genes.type,and 3 SNP sites were moderately polymorphic(0.25 < PIC < 0.50).Allele and genotype frequency analysis showed that all three SNP loci deviate from Hardy-Weinberg equilibrium(HWE),suggesting that these loci are in dynamic equilibrium under artificial breeding,migration and genetic drift.After correlating the growth traits and meat quality traits of Qinchuan cattle,it was found that the three SNPs in the promoter region of the FGF10 gene were significantly correlated with the growth traits and meat quality traits of Qinchuan beef cattle(P<0.05).Candidate marker genes for genetic improvement of size and carcass quality traits.The above results indicate that FGF10 has an inhibitory effect on the differentiation of bovine preadipocytes,and the specific role of FGF10 still needs further research to prove.The results of this study provide important data support for further elucidating the molecular mechanism of FGF10 gene regulation of bovine preadipocyte differentiation.