Characterization of the Methylation Status of <i>Pax7</i> and Myogenic Regulator Factors in Cell Myogenic Differentiation

Abstract

Epigenetic processes in the development of skeletal muscle have been appreciated for over a decade. DNA methylation is a major epigenetic modification important for regulating gene expression and suppressing spurious transcription. Up to now, the importance of epigenetic marks in the regulation of Pax7 and myogenic regulatory factors (MRFs) expression is far less explored. In the present study, semi-quantitative the real-time polymerase chain reaction (RT-PCR) analyses showed MyoD and Myf5 were expressed in activated and quiescent C2C12 cells. MyoG was expressed in a later stage of myogenesis. Pax7 was weakly expressed in differentiated C2C12 cells. To further understand the regulation of expression of these genes, the DNA methylation status of Pax7, MyoD, and Myf5 was determined by bisulfite sequencing PCR. During the C2C12 myoblasts fusion process, the changes of promoter and exon 1 methylation of Pax7, MyoD, and Myf5 genes were observed. In addition, an inverse relationship of low methylation and high expression was found. These results suggest that DNA methylation may be an important mechanism regulating Pax7 and MRFs transcription in cell myogenic differentiation.

Epigenetic regulation of satellite cell fate during skeletal muscle regeneration

“…The diminution of methylation at Myog promoter may lead to ZBT38 removal and abrogation of its repression [ 75 ]. In addition to Myog promoter demethylation, a general diminution of DNA methylation at the CpG sites of Myod1 promoter occurs after 3 days of differentiation in C2C12 cells [ 111 ]. DNA methylation changes during differentiation are not only attributed to demethylation of the promoter of genes required for myogenic differentiation.…”

“…In C2C12 cells, after 3 days of differentiation, an increase in DNA methylation of CpG sites of Pax7 and Myf5 promoters was shown. Interestingly, after 5 days of differentiation, the DNA methylation is still higher as compared with the proliferative state but slightly reduced as compared to day 3 of differentiation [ 111 ]. Genetic deletion of DNMT3A in mice resulted in fibrosis and reduction of cross-section area of the muscle after regeneration from an acute injury.…”

Efeitos de uma sessão de treinamento concorrente nos níveis de metilação e expressão de genes fatores regulatórios miogênicos

“…A metilação do DNA é um processo orquestrado por um conjunto de enzimas catalizadoras O entendimento desse mecanismo se desenvolveu nos últimos anos devido à grande quantidade de trabalhos científicos que o investigaram (HUPKES et al, 2011;BARRES et al, 2012;NITERT et al, 2012;CHAO et al, 2016;KING-HIMMELREICH et al, 2016;BEGUE et al, 2017;LAKER et al, 2017;SEABORNE et al, 2018). Com os avanços tecnológicos, uma série de métodos de análise da metilação do DNA foi desenvolvida e permitiu a explicação cada vez mais detalhada e fidedigna desse mecanismo epigenético (KARIMI, JOHANSSON, STACH, et al, 2006;SHEN & WATERLAND, 2007;HARRIS et al, 2010;LISANTI et al, 2013;KURDYUKOV & BULLOCK, 2016).…”

“…Especificamente quanto ao músculo esquelético, sabe-se que a metilação do DNA é um importante regulador da transcrição dos MRFs e do PAX7 (CHAO et al, 2016) (LAKER & RYALL, 2016. Alguns trabalhos com modelos celulares (e.g.…”