WGBS of embryonic gonads revealed that long non-coding RNAs in the MHM region might be involved in cell autonomous sex identity and female gonadal development in chickens.

IF 2.9 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Epigenetics Pub Date : 2024-12-01 Epub Date: 2023-12-01 DOI:10.1080/15592294.2023.2283657
Ligen Chen, Yu Cheng, Guixin Zhang, Yang Zhou, Zhen Zhang, Qianhong Chen, Yanping Feng
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引用次数: 0

Abstract

DNA methylation plays a key role in sex determination and differentiation in vertebrates. However, there are few studies on DNA methylation involved in chicken gonad development, and most focused on male hypermethylated regions (MHM). It is unclear whether there are specific differentially methylated regions (DMRs) in chicken embryonic gonads regulating sex determination and differentiation. Here, the DNA methylation maps showed that the difference of DNA methylation level between sexes was much higher at embryonic day 10 (E10) than that at embryonic day 6 (E6), and the significant differentially methylated regions at both stages were mainly distributed on the Z chromosome, including MHM1 and MHM2. The results of bisulphite sequencing PCR (BSP) and qRT-PCR showed hypomethylation of female MHM and upregulation of long non-coding RNAs (lncRNAs) whose promoter in the MHM region was consistent with the sequencing results, and similar results were in brain and muscle. In female sex-reversed gonads, the methylation pattern of MHM remained unchanged, and the expression levels of the three candidate lncRNAs were significantly decreased compared with those in females, but were significantly increased compared to males. The fluorescence in situ hybridization (FISH) results also showed that these lncRNAs were highly expressed in female embryonic gonads. The results of methyltransferase inhibitor and dual-luciferase reporter assay suggest that lncRNA expression may be regulated by DNA methylation within their promoters. Therefore, we speculated that MHM may be involved in cell-autonomous sex identity in chickens, and that lncRNAs regulated by MHM may be involved in female sexual differentiation.

胚胎性腺的WGBS表明,MHM区域的长链非编码rna可能参与了鸡细胞自主性别认同和雌性性腺发育。
DNA甲基化在脊椎动物的性别决定和分化中起着关键作用。然而,关于鸡性腺发育过程中DNA甲基化的研究很少,大多集中在雄性高甲基化区(MHM)。目前尚不清楚鸡胚性腺中是否存在调节性别决定和分化的特异性差异甲基化区(DMRs)。DNA甲基化图谱显示,胚胎第10天(E10)两性DNA甲基化水平差异远大于胚胎第6天(E6),且两期显著差异甲基化区域主要分布在Z染色体上,包括MHM1和MHM2。亚硫酸盐测序PCR (BSP)和qRT-PCR结果显示女性MHM低甲基化和长链非编码rna (lncRNAs)上调,其启动子在MHM区域与测序结果一致,脑和肌肉中也有类似的结果。在雌性性腺中,MHM的甲基化模式保持不变,3种候选lncrna的表达水平与雌性相比显著降低,但与雄性相比显著升高。荧光原位杂交(FISH)结果也显示这些lncrna在雌性胚胎性腺中高度表达。甲基转移酶抑制剂和双荧光素酶报告基因实验结果表明,lncRNA的表达可能受其启动子内DNA甲基化的调控。因此,我们推测MHM可能参与了鸡的细胞自主性别认同,MHM调控的lncrna可能参与了雌性性别分化。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Epigenetics
Epigenetics 生物-生化与分子生物学
CiteScore
6.80
自引率
2.70%
发文量
82
审稿时长
3-8 weeks
期刊介绍: Epigenetics publishes peer-reviewed original research and review articles that provide an unprecedented forum where epigenetic mechanisms and their role in diverse biological processes can be revealed, shared, and discussed. Epigenetics research studies heritable changes in gene expression caused by mechanisms others than the modification of the DNA sequence. Epigenetics therefore plays critical roles in a variety of biological systems, diseases, and disciplines. Topics of interest include (but are not limited to): DNA methylation Nucleosome positioning and modification Gene silencing Imprinting Nuclear reprogramming Chromatin remodeling Non-coding RNA Non-histone chromosomal elements Dosage compensation Nuclear organization Epigenetic therapy and diagnostics Nutrition and environmental epigenetics Cancer epigenetics Neuroepigenetics
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