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全转录组范围内 N6-甲基腺苷(m6A)图谱分析揭示了 m6A 在牦牛卵巢中的调控作用。

The transcriptome-wide N6-methyladenosine (mA) map profiling reveals the regulatory role of mA in the yak ovary.

机构信息

Key Laboratory of Yak Breeding Engineering of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou, 730050, China.

出版信息

BMC Genomics. 2022 May 11;23(1):358. doi: 10.1186/s12864-022-08585-7.

Abstract

BACKGROUND AND AIM

Yak estrus is a seasonal phenomenon, probably involving epigenetic regulation of synthesis and secretion of sex hormones as well as growth and development of follicles. N6-methyladenosine (mA) is the most common internal modification of the eukaryotic mRNA. However, there are no detailed reports on the mA transcriptome map of yak ovary. Therefore, this study aimed to collected the yak ovarian tissues at three different states of anestrus (YO-A), estrus (YO-F), and pregnancy (YO-P), and obtained the full transcriptome mA map in yak by MeRIP-seq.

RESULTS

The HE staining revealed that the number of growing follicles and mature follicles in the ovary during the estrus period was relatively higher than those in the anestrus period and the pregnancy period. The RT-qPCR showed that the expression of METTL3, METTL14, FTO, YTHDC1 were significantly different across different periods in the ovaries, which suggests that mA may play a regulatory role in ovarian activity. Next, we identified 20,174, 19,747 and 13,523 mA peaks in the three ovarian samples of YO-A, YO-F and YO-P using the methylated RNA immunoprecipitation sequencing (MeRIP-seq). The mA peaks are highly enriched in the coding sequence (CDS) region and 3'untranslated region (3'UTR) as well as the conserved sequence of "RRACH." The GO, KEGG and GSEA analysis revealed the involvement of mA in many physiological activities of the yak's ovary during reproductive cycle. The association analysis found that some genes such as BNC1, HOMER1, BMP15, BMP6, GPX3, and WNT11 were related to ovarian functions.

CONCLUSIONS

The comparison of the distribution patterns of methylation peaks in the ovarian tissues across different periods further explored the mA markers related to the regulation of ovarian ovulation and follicular development in the yak ovary. This comprehensive map provides a solid foundation for revealing the potential function of the mRNA mA modification in the yak ovary.

摘要

背景与目的

牦牛发情是季节性现象,可能涉及性激素合成和分泌以及卵泡生长和发育的表观遗传调控。N6-甲基腺苷(m6A)是真核 mRNA 最常见的内部修饰。然而,关于牦牛卵巢的 m6A 转录组图谱尚无详细报道。因此,本研究旨在采集处于乏情期(YO-A)、发情期(YO-F)和妊娠期(YO-P)的牦牛卵巢组织,通过 MeRIP-seq 获得牦牛卵巢全转录组 m6A 图谱。

结果

HE 染色显示发情期卵巢中生长卵泡和成熟卵泡的数量相对高于乏情期和妊娠期。RT-qPCR 显示卵巢中不同时期 METTL3、METTL14、FTO、YTHDC1 的表达存在显著差异,表明 m6A 可能在卵巢活动中发挥调节作用。接下来,我们使用甲基化 RNA 免疫沉淀测序(MeRIP-seq)在三个卵巢样本 YO-A、YO-F 和 YO-P 中鉴定出 20174、19747 和 13523 个 m6A 峰。m6A 峰高度富集在编码序列(CDS)区和 3'非翻译区(3'UTR)以及“RRACH”保守序列中。GO、KEGG 和 GSEA 分析表明 m6A 参与了牦牛卵巢生殖周期中许多生理活动。关联分析发现 BNC1、HOMER1、BMP15、BMP6、GPX3 和 WNT11 等一些基因与卵巢功能有关。

结论

不同时期卵巢组织中甲基化峰分布模式的比较进一步探讨了与牦牛卵巢排卵和卵泡发育调节相关的 m6A 标志物。该综合图谱为揭示牦牛卵巢中 mRNA m6A 修饰的潜在功能提供了坚实的基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7765/9092806/624cd13321af/12864_2022_8585_Fig1_HTML.jpg

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