Effect Of Akirin2 On Slow Myosin Heavy Chain Expression Through Arginine In Porcine Skeletal Muscle Satellite Cells

Akirin2 is a recently discovered gene that affects the meat quality,it is deemed to closely relate to muscle growth and marbling,suggesting Akirin2 may be a candidate gene for meat quality.Moreover,Arginine(Arg)had also closely related to muscle growth and intramuscular fat deposition.Therefore,this study aimed at revealing the effect of Akirin2 on slow myosin heavy chain expression in porcine skeletal muscle satellite cells and whether this effect is regulated by Arg.This study explored the effect of Akirin2 on slow myosin heavy chain expression in porcine skeletal muscle satellite cells by Akirinl overexpression and interference,and also explored the possible mechanisms of Akirin2 on slow myosin heavy chain expression by adding CsA and small-interfering RNA of NFATc1.In addition,this study was to explore whether the effect of Akirin2 on slow myosin heavy chain expression is regulated by Arg in porcine skeletal muscle satellite cells by adding Arg and small-interfering RNA of Akirin2.The main research concents and results are as follows:1.Effect of Akirin2 on slow myosin heavy chain expression in porcine skeletal muscle satellite cellsThe porcine skeletal muscle satellite cells were isolated by sterile dissection following collagenase digestion.The cell morphology was examined.Then the marker genes of the cells were detectd by immunofluorescence technology.In addition,we also explore the effect of overexpression and siRNA interference of the Akirin2 on slow myosin heavy chain expression.The results showed that skeletal muscle satellite cells of separation exhibited a typical long fusiform,and expressed the marker gene of Pax7.After inducing to the myotubes,MyHC showed positive expression.Overexpression of Akirin2 significantly increased MyHC I expression,and silencing of Akirin2 got opposite results.These findings revealed that Akirin2 could significantly increase slow myosin heavy chain expression.2.Possible mechanisms of Akirin2 on slow myosin heavy chain expression in porcine skeletal muscle satellite cellsThis study explored the effect of Akirin2 on NFATc1 and MCIP1.4 expression and CaN activity in porcine skeletal muscle satellite cells.through Akirin2 overexpression and interference,and further explored the possible mechanisms of Akirin2 on slow myosin heavy chain expression by adding CsA and small-interfering RNA of NFATcl.The results showed that overexpression of Akirin2 significantly increased NFATc1 and MCIP1.4 expression and CaN activity,and silencing of Akirin2 got opposite results.CsA significantly attenuated Akirin2 overexpression-induced upregulation of NFATc1 and MyHC I expressions and increase in the CaN activity.In addition,knockdown of NFATc1 abolished Akirin2 overexpression-induced upregulation of MyHC I.These findings revealed that Akirin2 regulates slow myosin heavy chain expression by CaN-NFATc1 signaling in porcine skeletal muscle satellite cells.3.Effect of Akirin1 on slow myosin heavy chain expression through Arg in porcine skeletal muscle satellite cellsThis study explored the effect of Arg on slow myosin heavy chain expression in porcine skeletal muscle satellite cells,and further explored whether the effect of Akirin1 on slow myosin heavy chain expression is regulated by Arg in porcine skeletal muscle satellite cells by adding Arg and small-interfering RNA of Akirin2.The results showed that Arg significantly increased MyHC I and Akirin2 expression.However,knockdown of Akirin1 abolished Arg-induced upregulation of MyHC I.These findings revealed that Akirin2-induced slow myosin heavy chain expression is regulated by Arg in porcine skeletal muscle satellite cells.In summary,this study proved that Akirin2 regulates slow myosin heavy chain expression by CaN-NFATcl signaling in porcine skeletal muscle satellite cells,and Akirin2-induced slow myosin heavy chain expression is regulated by Arg in porcine skeletal muscle satellite cells.This study will lay an important molecular basis for discussing the regulation mechanism of pork quality.Meanwhile,this study will also provide a new idea to improve animal meat quality by taking nutritional strategies.