Preliminary Study Of Laying Hens Broiler Muscle Cells Differentially Expressed Proteins Identification And Oxct1 Gene Function

Investigation of myogenesis is very important for understanding the whole process ofembryonic development. To give an insight into the molecular mechanism of myogenesis,broiler and layer chicken have been utilized as animal models because of theirsignificant difference in skeletal muscle growth rate. And 2-D method was employed tofind the difference between the broiler and layer myoblast. As the results, 8 differentialspots including OXCT1 protein; capping protein (actin filament) muscle Z-line, alpha 2;PREDICTED: similar to Rbm6 protein; PREDICTED: similar to prohibitin [Gallusgallus]; tumor protein, translationally-controlled 1; acidic ribosomal phosphoprotein;PREDICTED: similar to Dihydroxyacetone and phosphate acyltransferase (DHAP) werefound expressed higher in broiler myoblast than layer's.The real-time PCR confirm results showed that 4 proteins: OXCT1 protein;PREDICTED: similar to Rbm6 protein; PREDICTED: similar to calmodulin-like 4 andtumor protein, translationally-controlled 1 also expressed higher in broiler myoblast thanlayer's on mRNA level.OXCT1 (3-oxoacid CoA transferase 1) which mediates the rate-determining step ofketolysis in extrahepatic tissues is a key enzyme for ketone body catabolism. As the cellsgrowth relying on the cell metabolism, we supposed that the OXCT1 difference may berelated to broiler and layer's skeletal muscle big difference. To further study the functionof OXCT1 gene, polyantibody of preparation was studied. And the polyantibody couldrecognize OXCT1 protein from chicken, mouse and human tissues specifically.We investigated the OXCT1 of the broiler and layer's different stage' skeletalmuscle on the mRNA level, protein level and enzyme activity level and found that theOXCT1 expressed much higher in broiler than in layer at all the chicken growth stages. To find the relationship between the OXCT1 and the ketone-body, the serum 3-HBlevel of the different chicken stages were detected and the results showed that the 3-HBlevel was higher in layer than in broiler which was not consist to the OXCT1 expressionlevel in chicken skeletal muscle. These results implied that broiler might use moreketone-body in chicken embryo stage.The test of OXCT1 overexpression and the OXCT1 RNAi in broiler were done tofind whether the OXCT1 would have effect on the cell phenotype change. Although thecell phenotype had no change but the FACS results showed that overexpression ofOXCT1 in broiler myoblast could increase the DNA synthsis.Collectively, these results suggested that skeletal muscle myoblast from broiler andlayer could be used as an excellent in vivo system to find the differential proteinsbetween broiler and layer using the proteomics 2-D method. And studying thedifferential expressed proteins in chicken myoblast would help us to explain the bigdifference between broiler and layer myogenesis. Also it might give us some clues aboutthe organ size control.