Earlier demethylation of myogenic genes contributes to embryonic precocious terminal differentiation of myoblasts in miniature pigs

Zhang, Xumeng; Nie, Yaping; Cai, Shaohui; Ding, Suying; Fu, Bingqiang; Wang, Hong; Chen, Luxi; Liu, Xiaohong; Ming-gui, Liu; Yuan, Renqiang; Qiu, Boqin; He, Zuyong; Peng, Cong; Chen, Yaosheng; Mo, Delin

doi:10.1096/fj.201900388r

Cited by 13 publications

(14 citation statements)

References 70 publications

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“…There were more eMyHC + cells in LT ( Fig. 1a and b), which is consistent with previous reports, indicating precocious terminal differentiation of myoblasts in LT [20]. To de ne the progression of myogenic differentiation, dual immunostaining of Pax7 and MyoD was conducted.…”

Section: Resultssupporting

confidence: 90%

“…Different pig breeds vary in muscle mass because of differences in muscle development. In recent years, several studies revealed that pigs with less muscle showed earlier formation of primary myo bers, which arise from earlier expression of myogenic genes [20]. In the present report, we found more eMyHC-positive cells in LT LDM at 35 dpc, indicating precocious terminal differentiation of myoblasts in LT. MyoD, which can initiate the process of multiple non-muscle cell lineages into muscle cell lineages, is a crucial master switch in regulating muscle-speci c gene transcription [27,28].…”

Section: Discussionsupporting

confidence: 72%

“…For example, the myo ber density and diameter were signi cantly higher in Meishan pigs (fatty, a Chinese indigenous breed) compared to Large White pigs (a lean breed) at 35dpc [19]. In our previous study, we found that Wuzhishan pigs (fatty) showed earlier myoblast differentiation than that of Landrace pigs (LR, lean) in embryonic stages [20]. Primary muscle bers appear earlier in Lantang (LT, fatty) than that in LR, at 35dpc and 42dpc respectively [21].…”

Section: Introductionmentioning

confidence: 83%

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Notch Signaling Slows Down the Progression of Embryonic Myogenic Differentiation in Landrace

Cai¹,

Zhu²,

Bin³

et al. 2020

Preprint

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BackgroundDelving into porcine embryonic myogenesis is the key to elucidating the complex regulation of breed-specific differences in growth performance and meat production. Increasing evidence proved that pigs with less meat production showed more intense embryonic myogenesis, but little is known about the underlying mechanisms. ResultsIn this study, we confirmed that the differentiation process of myogenic progenitors in Lantang pig (LT, fat) was faster than that in Landrace pig (LR, lean), which resulted in more differentiated myoblasts (Pax7-/MyoD+) but less myogenic progenitors (Pax7+/MyoD-) in LT at 35 days post-conception (35dpc). Additionally, in vitro, embryonic myogenic progenitors isolated from LT showed stronger differentiation capacity with earlier expression of MyoD. Furthermore, the expression levels of genes related to Notch signaling in LR progenitor cells were significantly higher than that of LT, while there was no significant difference between the two breeds in gene expression levels of Wnt and Akt/mTOR pathway. Inhibition of Notch signaling or knockdown of Pax7 promotes myogenic differentiation of primary progenitor cells or myoblasts, while activation of Notch signaling or overexpression of Pax7 has the opposite effects.ConclusionsMyogenic differentiation is more rapid in LT than that in LR at 35dpc. Mechanically, Notch signaling facilitates maintenance of myogenic progenitor cells and antagonizes myogenic differentiation by promoting Pax7 expression but preventing MyoD expression in LR.

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Section: Resultssupporting

confidence: 90%

Section: Discussionsupporting

confidence: 72%

Section: Introductionmentioning

confidence: 83%

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Notch Signaling Slows Down the Progression of Embryonic Myogenic Differentiation in Landrace

Cai¹,

Zhu²,

Bin³

et al. 2020

Preprint

Self Cite

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“…We found that on day 28, overall DNA methylation, as evaluated from the median methylation of 100-CpG tiles, was 15.38% in male germ cells and 15.85% in female germ cells. Given that the methylation level in somatic cells of the pig fetus on day 28 has been reported at around 75% [25], it seems the main wave of DNA demethylation occurred in the germ cells before day 28 (Fig. 2a).…”

Section: Dynamic Methylome Analysis Of Porcine Germ Cellsmentioning

confidence: 96%

Genome-wide DNA methylation dynamics during epigenetic reprogramming in the porcine germline

Gómez-Redondo¹,

Planells²,

Cánovas

et al. 2021

Clin Epigenet

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Background Prior work in mice has shown that some retrotransposed elements remain substantially methylated during DNA methylation reprogramming of germ cells. In the pig, however, information about this process is scarce. The present study was designed to examine the methylation profiles of porcine germ cells during the time course of epigenetic reprogramming. Results Sows were artificially inseminated, and their fetuses were collected 28, 32, 36, 39, and 42 days later. At each time point, genital ridges were dissected from the mesonephros and germ cells were isolated through magnetic-activated cell sorting using an anti-SSEA-1 antibody, and recovered germ cells were subjected to whole-genome bisulphite sequencing. Methylation levels were quantified using SeqMonk software by performing an unbiased analysis, and persistently methylated regions (PMRs) in each sex were determined to extract those regions showing 50% or more methylation. Most genomic elements underwent a dramatic loss of methylation from day 28 to day 36, when the lowest levels were shown. By day 42, there was evidence for the initiation of genomic re-methylation. We identified a total of 1456 and 1122 PMRs in male and female germ cells, respectively, and large numbers of transposable elements (SINEs, LINEs, and LTRs) were found to be located within these PMRs. Twenty-one percent of the introns located in these PMRs were found to be the first introns of a gene, suggesting their regulatory role in the expression of these genes. Interestingly, most of the identified PMRs were demethylated at the blastocyst stage. Conclusions Our findings indicate that methylation reprogramming in pig germ cells follows the general dynamics shown in mice and human, unveiling genomic elements that behave differently between male and female germ cells.

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“…Additionally, previous studies have demonstrated that TETs play important roles in the regulation of muscle development [27][28][29] via the demethylation of myogenesis-specific genes. TET1 and TET2 were found to meditate the conversion of methylated cytosine into 5hmC, which is the first step in the DNA demethylation active pathway [30][31][32].…”

Section: Discussionmentioning

confidence: 99%

MSTN Mutant Promotes Myogenic Differentiation by Increasing Demethylase TET1 Expression via the SMAD2/SMAD3 Pathway

et al. 2020

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Myostatin (MSTN) is mostly expressed in skeletal muscle and plays crucial roles in the negative regulation of muscle mass development. The methylation and demethylation of myogenesis-specific genes are major regulatory factors in muscle satellite cell differentiation. The present study was designed to investigate the mechanism of myogenic differentiation regulated by MSTN mutation (MT) and the methylation/demethylation state of downstream genes. The results showed that, in the MSTN-/+ satellite cells, a higher myotube fusion index and a larger myotube length were observed compared to the wild type controls; the genes associated with myogenesis were all up-regulated compared to the WT controls. The methylation of the promoters and gene bodies of PAX3, PAX7, MyoD, and MyoG were all down-regulated, while the expression of the key demethylase TET1 was significantly promoted. ChIP-qPCR was used to demonstrate that the SMAD2/SMAD3 complex combined with the promoter of TET1 to inhibit the activity of TET1 promoter, indicating that MSTN may regulate TET1 via SMAD2/SMAD3. The overexpression of TET1 in wild type cells promoted myogenic differentiation, increased the myotube index, and reduced the methylation of the associated genes. On the contrary, the knockdown of TET1 in the MSTN mutant cells resulted in the opposite phenomena as in the overexpressed cells. In conclusion, the myostatin mutant showed an increased transcriptional activity of TET1, inducing higher levels of demethylation and improving the transcriptional activity levels of myogenic differentiation-associated genes. The binding of SMAD2/SMAD3 directly to the TET1 promoter region indicated that the MSTN mutant demethylated the myogenesis-specific genes by up-regulating TET1, which is directly controlled by SMAD2/SMAD3.

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Earlier demethylation of myogenic genes contributes to embryonic precocious terminal differentiation of myoblasts in miniature pigs

Cited by 13 publications

References 70 publications

Notch Signaling Slows Down the Progression of Embryonic Myogenic Differentiation in Landrace

Notch Signaling Slows Down the Progression of Embryonic Myogenic Differentiation in Landrace

Genome-wide DNA methylation dynamics during epigenetic reprogramming in the porcine germline

MSTN Mutant Promotes Myogenic Differentiation by Increasing Demethylase TET1 Expression via the SMAD2/SMAD3 Pathway

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