Comprehensive Analysis of the Transcriptome-Wide m<sup>6</sup>A Methylome in Shaziling Pig Testicular Development
2023
Chujie Chen | Xiangwei Tang | Saina Yan | Anqi Yang | Jiaojiao Xiang | Yanhong Deng | Yulong Yin | Bin Chen | Jingjing Gu
RNA <i>N</i><sup>6</sup>-methyladenosine (m<sup>6</sup>A) modification is one of the principal post-transcriptional modifications and plays a dynamic role in testicular development and spermatogenesis. However, the role of m<sup>6</sup>A in porcine testis is understudied. Here, we performed a comprehensive analysis of the m<sup>6</sup>A transcriptome-wide profile in Shaziling pig testes at birth, puberty, and maturity. We analyzed the total transcriptome m<sup>6</sup>A profile and found that the m<sup>6</sup>A patterns were highly distinct in terms of the modification of the transcriptomes during porcine testis development. We found that key m<sup>6</sup>A methylated genes (<i>AURKC</i>, <i>OVOL</i>, <i>SOX8</i>, <i>ACVR2A</i>, and <i>SPATA46</i>) were highly enriched during spermatogenesis and identified in spermatogenesis-related KEGG pathways, including Wnt, cAMP, mTOR, AMPK, PI3K-Akt, and spliceosome. Our findings indicated that m<sup>6</sup>A methylations are involved in the complex yet well-organized post-transcriptional regulation of porcine testicular development and spermatogenesis. We found that the m<sup>6</sup>A eraser <i>ALKBH5</i> negatively regulated the proliferation of immature porcine Sertoli cells. Furthermore, we proposed a novel mechanism of m<sup>6</sup>A modification during testicular development: <i>ALKBH5</i> regulated the RNA methylation level and gene expression of <i>SOX9</i> mRNA. In addition to serving as a potential target for improving boar reproduction, our findings contributed to the further understanding of the regulation of m<sup>6</sup>A modifications in male reproduction.
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