Wang Yushi, Yan Xiping, Liu Hehe, Hu Shenqiang, Hu Jiwei, Li Liang, Wang Jiwen
Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu 611130, PR China.
Agriculture Bureau of Ya'an, Ya'an 625000, PR China.
J Therm Biol. 2019 Feb;80:75-81. doi: 10.1016/j.jtherbio.2018.12.023. Epub 2018 Dec 26.
Avian embryos are an ideal system to investigate the effect of incubation temperature on embryonic development, but the characteristics and mechanisms of temperature effects on poultry embryonic myogenesis are unclear. In this study, we investigated the effect of increasing the incubation temperature by 1 °C on the expression of nine myogenesis-related genes in ducks and then explored the correlation between the alteration of promoter methylation and the expression of two of the nine genes under thermal manipulation (TM). The qRT-PCR results showed that TM during embryonic days (ED) 1-10 promoted (P < 0.05) the expression of genes in breast muscle (PAX3, PAX7, MYOG, MCK, SIX1, TNNC1) and leg muscle (MYOD, MYOG, MYF5, MCK, AKIRIN2, TNNC1). TM during ED10-20 promoted the expression of PAX3, MYF5 and MCK and inhibited AKIRIN2 expression in breast muscle (P < 0.05); however, it inhibited the expression of PAX3, PAX7, MYOD, MYOG, MYF5, SIX1, AKIRIN2 and TNNC1 and promoted MCK expression in leg muscle (P < 0.05). TM during ED20-27 inhibited the expression of genes in breast muscle (PAX7) and leg muscle (MYOD, MYOG, MYF5, TNNC1) and promoted MCK expression in breast and leg muscle (P < 0.05). Furthermore, with the Sequenom MassARRAY platform, it was observed that the average methylation level of AKIRIN2 (ED10) and TNNC1 (ED20) in leg muscle decreased (P < 0.05) after TM. Notably, we found significant (P < 0.05) inverse correlations between the methylation and mRNA levels of AKIRIN2 under TM during ED1-10 (r = - 0.969) and ED10-20 (r = - 0.805). Taken together, TM during ED1-10 was more favorable for improving duck myogenesis-related gene expression than TM during ED10-20 and ED20-27. TM during duck embryogenesis seemed to have a greater effect on the development of leg muscle than breast muscle and might alter AKIRIN2 expression by changing its promoter methylation status. These findings may be helpful to understand temperature effects on the muscle development of avian embryos and to explore the role of epigenetic regulation during this process.
禽类胚胎是研究孵化温度对胚胎发育影响的理想系统，但温度对家禽胚胎肌生成的影响特征和机制尚不清楚。在本研究中，我们研究了将孵化温度提高1°C对鸭9个肌生成相关基因表达的影响，然后探讨了启动子甲基化改变与热调控（TM）下9个基因中2个基因表达之间的相关性。qRT-PCR结果显示，胚胎期（ED）1-10天的TM促进（P<0.05）胸肌（PAX3、PAX7、MYOG、MCK、SIX1、TNNC1）和腿肌（MYOD、MYOG、MYF5、MCK、AKIRIN2、TNNC1）中基因的表达。ED10-20天的TM促进胸肌中PAX3、MYF5和MCK的表达并抑制AKIRIN2的表达（P<0.05）；然而，它抑制腿肌中PAX3、PAX7、MYOD、MYOG、MYF5、SIX1、AKIRIN2和TNNC1的表达并促进MCK的表达（P<0.05）。ED20-27天的TM抑制胸肌（PAX7）和腿肌（MYOD、MYOG、MYF5、TNNC1）中基因的表达并促进胸肌和腿肌中MCK的表达（P<0.05）。此外，使用Sequenom MassARRAY平台观察到，TM后腿肌中AKIRIN2（ED10）和TNNC1（ED20）的平均甲基化水平降低（P<0.05）。值得注意的是，我们发现ED1-10天（r=-0.969）和ED10-20天（r=-0.805）TM期间AKIRIN2的甲基化水平与mRNA水平之间存在显著的（P<0.05）负相关。综上所述，ED1-10天的TM比ED10-20天和ED20-27天的TM更有利于提高鸭肌生成相关基因的表达。鸭胚胎发育期间的TM对腿肌发育的影响似乎比对胸肌的影响更大，并且可能通过改变其启动子甲基化状态来改变AKIRIN2的表达。这些发现可能有助于理解温度对禽类胚胎肌肉发育的影响，并探索这一过程中表观遗传调控的作用。
