Effects of myogenin on muscle fiber types and key metabolic enzymes in gene transfer mice and C2C12 myoblasts.
Skeletal muscle fiber type composition is one of the important factors influencing muscle growth and meat quality. As a member of the myogenic transcription factors, myogenin (MyoG) is required for embryonic myoblast differentiation, but the expression of MyoG continues in mature muscle tissue of adult animals, especially in oxidative metabolic muscle, which suggests that MyoG may play a more extended role. Therefore, using MyoG gene transfer mice and C2C12 myoblasts as in vivo and in vitro models, respectively, we elected to study the role of MyoG in muscle fiber types and oxidative metabolism by using overexpression and siRNA suppression strategies. The overexpression of MyoG by DNA electroporation in mouse gastrocnemius muscle had no significant effect on fiber type composition but upregulated the mRNA expression (P<0.01) and enzyme activity (P<0.05) of oxidative succinic dehydrogenase (SDH). In addition, downregulation of the activity of the glycolytic enzymes lactate dehydrogenase (LDH, P<0.05) and pyruvate kinase (PK, P<0.05) was observed in MyoG gene transfer mice. In vitro experiments verified the results obtained in mice. Stable MyoG-transfected differentiating C2C12 cells showed higher mRNA expression levels of myosin heavy chain (MyHC) isoform IIX (P<0.01) and SDH (P<0.05), while the LDH mRNA was attenuated. The enzyme activities of SDH (P<0.01) and LDH (P<0.05) were similarly altered at the mRNA level. When MyoG was knocked down in C2C12 cells, MyHC IIX expression (P<0.05) was decreased, but the mRNA level (P<0.05) and the enzyme activity (P<0.05) of SDH were increased. Downregulating MyoG also increased the activity of the glycolytic enzymes PK (P<0.05) and hexokinase (HK, P<0.05). Based on those results, we concluded that MyoG barely changes the MyHC isoforms, except MyHC IIX, in differentiating myoblasts but probably influences the shift from glycolytic metabolism towards oxidative metabolism both in vivo and in vitro. These results contribute to further understand the role of MyoG in skeletal muscle energy metabolism and also help to explore the key genes that regulate meat quality.
Institute of Feed Science, Zhejiang University, The Key Laboratory of Molecular Animal Nutrition, Ministry of Education, Zhejiang Provincial Laboratory of Feed and Animal Nutrition, Hangzhou, Zhejiang, PR China.
Lin-Na
LN
Zhu
Yang
Y
Ren
Jing-Qing
JQ
Chen
Yi-Zhen
YZ
Wang
D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D063427:Food Quality; D015870:Gene Expression; D015971:Gene Expression Regulation, Enzymologic; D055785:Gene Knockdown Techniques; D018014:Gene Transfer Techniques; D006019:Glycolysis; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008291:Malate Dehydrogenase / Q000378:metabolism; D008297:Male; D008460:Meat / Q000592:standards; D051379:Mice; D008813:Mice, Inbred ICR; D008931:Mitochondria, Muscle / Q000378:metabolism; D018485:Muscle Fibers, Skeletal / Q000201:enzymology* / Q000378:metabolism; D018482:Muscle, Skeletal / Q000166:cytology / Q000201:enzymology; D032446:Myoblasts / Q000201:enzymology* / Q000378:metabolism / Q000502:physiology; D018008:Myogenin / Q000096:biosynthesis / Q000235:genetics*; D018995:Myosin Heavy Chains / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D013385:Succinate Dehydrogenase / Q000378:metabolism; D034421:Sus scrofa
C503801:Myog protein, mouse; D018008:Myogenin; D034741:RNA, Small Interfering; D007770:L-Lactate Dehydrogenase; D008291:Malate Dehydrogenase; D013385:Succinate Dehydrogenase; D018995:Myosin Heavy Chains
HK; LDH; Muscle fiber types; MyHC; MyoG; Myogenin; Oxidative metabolism; PK; SDH; hexokinase; lactate dehydrogenase; myogenin; myosin heavy chain; pyruvate kinase; succinic dehydrogenase