Effect of Long Non-coding RNA MSTRG.14200 on Porcine Skeletal Muscle Satellite Cell Differentiation and Muscle Fiber Transformation
[Objective]This experiment was conducted to investigate the effects of long non-coding RNA(lncRNA)MSTRG.14200 on myogenic differentiation of porcine skeletal muscle satellite cells and the transformation of different types of muscle fibers.[Method]Three 6-month-old Duroc pigs were slaughtered,and the heart,liver,spleen,lung,kidney,stomach,longissimus dorsi muscle,leg muscle,soleus and extensor digitorum longus were collected.Real-time quantitative PCR was used to detect the expression of MSTRG.14200 in different tissues.The MSTRG.14200 overexpression vector pcDNA3.1-14200 was constructed and transfected into PSCs cells,and the cells were collected after 3 days of induced differentiation.The mRNA and protein expressions of myosin heavy chain(MyHC),myogenin(MyoG),mydifferentiation factor(MyoD),MyHC Ⅰ,MyHC Ⅱ a,MyHC Ⅱ b and MyHC Ⅱ x were detected by Real-time quantitative PCR and Western blotting,respectively.The percentage of MyHC,MyHC Ⅰ and MyHC Ⅱ b positive cells was detected by immunofluorescence assay.[Result]MSTRG.14200 was expressed in different tissues of Duroc pigs,and the expression was the highest in longissimus dorsi muscle,which was extremely significantly higher than other tissues(P＜0.01),and there were significant differences in the expression of extensor digitorum longus and soleus(P＜0.05).After overexpression of MSTRG.14200,mRNA levels of MyoD and MyHC genes related to cell differentiation were extremely significantly increased(P＜0.01),and protein levels of MyoD,MyoG and MyHC were significantly or extremely significantly increased(P＜0.05 or P＜0.01),the percentage of MyHC positive cells was extremely significantly increased(P＜0.01).The mRNA and protein expressions of MyHC Ⅰ gene related to muscle fiber transformation and the proportion of positive cells were significantly or extremely significantly decreased(P＜0.05 or P＜0.01),the mRNA and protein expression of MyHC Ⅱ b gene and the percentage of positive cells were significantly increased(P＜0.05).[Conclusion]MSTRG.14200 could promote myogenic differentiation of porcine skeletal muscle and promote the transformation of slow muscle fibers into fast muscle fibers.The results provided a theoretical basis for exploring the molecular mechanism of transformation of porcine skeletal muscle fiber type.
万方数据
维普
