Biological Function Studies Of Candidate Genes S100A10 And Bta-miR-210 Related To Intramuscular Fat Deposition In Cattle

Intramuscular fat(IMF)content is a crucial factor affecting beef quality.In the fattening of beef cattle,the farmers expect to produce beef with abundant fat deposition in muscle and less feed consumption for higher profit.Therefore,it is of great significance to breed beef cattle with both high growth rate and IMF content.IMF is the adipose tissue that deposited between skeletal muscle fibers and muscle bundles.Adipose tissue content is determined by the number and size of adipocytes.Adipose tissue deposition and distribution are controled by genetic and nutritional factors.In order to explore the development mechanism of beef cattle IMF deposition,the mechanism of beef quality traits related gene S100A10 and bta-miR-210 in IMF development were studied by revealing their roles in the adipogenic differentiation in vitro.In mouse preadipocytes,S100A10 expression was knockdown by siRNA to study its effect on white/brown adipogenic differentiation.This study provides scientific basis for the study of S100A10 gene on IMF in beef cattle.Exogenous overexpression bta-miR-210 and target gene knockdown were used to investigate the effect on adipogenic differentiation of bovine PDGFR?~+progenitors.The main results are as follows:1 Preadipocytes were isolated from 12 week old mice by collagenase digestion.PDGFR?~+progenitors were isolated from skeletal muscle of bovine fetuses at 5 month old by collagenase digestion and cell-surface markers.2 White/brown adipogenic differentiation were induced when the mouse preadipocytes reached100%.Meanwhile,siRNA was used to interfere the expression of S100A10 gene.The lipid droplets were significantly reduced at sixth day of differentiation.The specific genes of adipogenic differentiation were significantly decreased by qRT-PCR,Western blot and immunofluorescence assay.3 During the differentiation of PDGFR?~+progenitors,the expression of bta-miR-210 was up-regulated.4 The lipid droplets were significantly increased at ninth day after bta-miR-210 overexpression.The specific genes of adipogenic differentiation were significantly increased.5.WISP2 was identified as a target gene of bta-miR-210 using dual luciferase report system.