Regulation Of CTRP3 On Myo Genic Differentiation Of Geese Skeletal Muscle Cells

Goose is an ideal high-protein,low-fat,low-cholesterol health food,which is favored by consumers.It has great effects to improve the meat production of goose.Exploring the functional genes that play an important role in the skeletal muscle tissue,and understanding the molecular mechanism of the regulation of goose skeletal muscle growth and development,are both vital to the improvement of the meat production performance of goose.CTRP3(C1q/tumor necrosis factor-related protein-3)is an important factor involved in muscle growth and development,and is closely related to the regulation of skeletal muscle cell differentiation,but its regulation in the development of goose has not been clarified.This study used goose as target and studied three parts in skeletal muscle cell differentiation in goose.(1)We isolated the skeletal muscle satellite cells in vitro by enzymatic digestion,and identified them using immunofluorescence of the specific marker genes of skeletal muscle satellite cells during proliferation.We inducted skeletal muscle satellite cells and established a cell differentiation model that is stably passaged in vitro.(2)The expression of CTRP3 gene in goose embryo development was detected by qRT-PCR,and the expression of CTRP3 gene in skeletal muscle satellite cells was detected.Overexpression vector of CTRP3 gene was constructed using eukaryotic expression vector pCDNA3.1.After overexpression and silencing CTRP3 gene,we checked the gene and protein expression level of the CTRP3 and its associated marker gene by qRT-PCR and Western blot.(3)The miRNAs that may target CTRP3 gene were predicted using bioinformatics prediction software.The predicted miRNA mimics were synthesized and the mi RNA mimics were transfected into skeletal muscle satellite cells,and the expression of candidate miRNAs and CTRP3 genes were detected by qRT-PCR and Western blot.Further speculation on the regulation of CTRP3 gene by candidate miRNAs.In this study,(1)we isolated the goose skeletal muscle satellite cells cultured them in vitro,and constructed an in vitro differentiation model for subsequent related experimental studies;(2)the expression profile of CTRP3 in goose skeletal muscle development was successfully obtained,and we proved CTRP3 can promote the differentiation of goose skeletal muscle satellite cells by overexpression and interference tests;(3)bioinformatics prediction combined with preliminary analysis of the experiment revealed that miR-129-5p may act on the goose CTRP3 gene.