Li Jianbo, Zhang Xiuan, Wang Xiqiong, Sun Congjiao, Zheng Jiangxia, Li Junying, Yi Guoqiang, Yang Ning
National Engineering Laboratory for Animal Breeding and Key Laboratory of Animal Genetics, Breeding and Reproduction, Ministry of Agriculture and Rural Affairs, China Agricultural University, Beijing, China.
Shenzhen Branch, Guangdong Laboratory of Lingnan Modern Agriculture, Genome Analysis Laboratory of the Ministry of Agriculture and Rural Affairs, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen, China.
J Anim Sci Biotechnol. 2022 May 18;13(1):52. doi: 10.1186/s40104-022-00710-6.
As a ubiquitous reversible epigenetic RNA modification, N6-methyladenosine (m6A) plays crucial regulatory roles in multiple biological pathways. However, its functional mechanisms in sex determination and differentiation during gonadal development of chicken embryos are not clear. Therefore, we established a transcriptome-wide m6A map in the female and male chicken left gonads of embryonic day 7 (E7) by methylated RNA immunoprecipitation sequencing (MeRIP-seq) to offer insight into the landscape of m6A methylation and investigate the post-transcriptional modification underlying gonadal differentiation.
The chicken embryonic gonadal transcriptome was extensively methylated. We found 15,191 and 16,111 m6A peaks in the female and male left gonads, respectively, which were mainly enriched in the coding sequence (CDS) and stop codon. Among these m6A peaks, we identified that 1013 and 751 were hypermethylated in females and males, respectively. These differential peaks covered 281 and 327 genes, such as BMP2, SMAD2, SOX9 and CYP19A1, which were primarily associated with development, morphogenesis and sex differentiation by functional enrichment. Further analysis revealed that the m6A methylation level was positively correlated with gene expression abundance. Furthermore, we found that YTHDC2 could regulate the expression of sex-related genes, especially HEMGN and SOX9, in male mesonephros/gonad mingle cells, which was verified by in vitro experiments, suggesting a regulatory role of m6A methylation in chicken gonad differentiation.
This work provided a comprehensive m6A methylation profile of chicken embryonic gonads and revealed YTHDC2 as a key regulator responsible for sex differentiation. Our results contribute to a better understanding of epigenetic factors involved in chicken sex determination and differentiation and to promoting the future development of sex manipulation in poultry industry.
作为一种普遍存在的可逆表观遗传RNA修饰，N6-甲基腺苷（m6A）在多种生物学途径中发挥着关键的调控作用。然而，其在鸡胚性腺发育过程中性别决定和分化的功能机制尚不清楚。因此，我们通过甲基化RNA免疫沉淀测序（MeRIP-seq）建立了胚胎第7天（E7）雌性和雄性鸡左侧性腺的全转录组m6A图谱，以深入了解m6A甲基化格局，并研究性腺分化的转录后修饰。
鸡胚性腺转录组广泛甲基化。我们分别在雌性和雄性左侧性腺中发现了15,191个和16,111个m6A峰，这些峰主要富集在编码序列（CDS）和终止密码子中。在这些m6A峰中，我们鉴定出分别有1013个和751个在雌性和雄性中发生了高甲基化。这些差异峰覆盖了281个和327个基因，如BMP2、SMAD2、SOX9和CYP19A1，通过功能富集分析，它们主要与发育、形态发生和性别分化相关。进一步分析表明，m6A甲基化水平与基因表达丰度呈正相关。此外，我们发现YTHDC2可以调节雄性中肾/性腺混合细胞中性别相关基因的表达，特别是HEMGN和SOX9，这一点通过体外实验得到了验证，表明m6A甲基化在鸡性腺分化中具有调控作用。
这项工作提供了鸡胚性腺全面的m6A甲基化图谱，并揭示YTHDC2是负责性别分化的关键调节因子。我们的结果有助于更好地理解参与鸡性别决定和分化的表观遗传因素，并促进家禽业性别控制技术的未来发展。
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