Proteomics Study On Erythrocytes Of MSTN Gene-edited Cattle

Myostatin(MSTN),a negative regulator of skeletal muscle growth and development,regulates proliferation and differentiation of myoblasts.The decrease in both of the expression and activity of MSTN protein leads to the increase in skeletal muscle mass.Exercise can reduce the expression of MSTN m RNA;MSTN knockout mice are muscle hypertrophy but easy to fatigue,yet exercise can weaken muscle hypertrophy and restore skeletal muscle function.The deficiency of MSTN would change the metabolic pathway of muscle energy,and exercise can induce the change of energy metabolic pathway of erythrocyte.In this study,the proteome of erythrocyte samples of 4 MSTN edited cattle(MT)and 4 wild type cattle(WT)before(Q)and after(H)exercise were studied to explore the relationship of MSTN,exercise and erythrocyth.The samples contained four groups MSTN edited cattle before exercise(MTQ),wild type cattle before exercise(WTQ),MSTN edited cattle after exercise(MTH)and wild type cattle after exercise(WTH).The number of identified proteins with quantitative in the label-free quantification proteomic value were 843,819,846,835 respectively.By comparing the difference of erythrocyte proteome among 8 cattle after and before exercise,the effect of exercise on bovine erythrocyte was analyzed.37 proteins were up-regulated while 1 protein was down.The up-regulated proteins are mainly related to proteasome and glycolysis and pentose phosphate pathway.26 S proteasome complex subunits contained ?1 subunit(PSMA6),?3 subunit(PSMA4),?1 subunit(PSMB6),?5 subunit(PSMB5)of 20 S core granules and Rpn1(PSMD2),Rpn2(PSMD1),Rpn3(PSMD3),Rpt6(PSMC5)of 19 S regulatory particles.The key enzymes of glycolysis and pentose phosphate pathway were PGM2,PFKM,PKLR,GALK1 and PRPS1.Studies have shown that exercise causes oxidative damage to erythrocytes,which produce more reductive NADPH to remove ROS by enhancing the pentose phosphate pathway,and mobilize proteasome to degrade damaged proteins for maintaining intracellular homeostasis.The difference of erythrocyte proteome between4 MTQ and 4 WTQ was compared to analyze the effect of MSTN edited on bovine erythrocytes.25 proteins were up-regulated and 88 proteins were down-regulated.The functions of these proteins are mainly related to the regulation of cell metabolism,biological stress,coagulation and hemostasis,which shows that the antibacterial ability and clotting ability of erythrocytes might be decreased,when MSTN of bovine was edited.By comparing the differences of erythrocyte protein changes between 4 MT and4 WT after and before exercise,the effect of MSTN edited on erythrocyte proteome of cattle before and after exercise were analyzed.9 proteins were up-regulated and 29 proteins were down-regulated.The main functions of these proteins are involved in the metabolism and biosynthesis of flavin nucleotides and the c GMP passway.The results suggested that the erythrocytes antioxidant capacity of MSTN edited bovine against oxidative damage caused by exercise would been enhanced.