DNA methylation modification is associated with gonadal differentiation in Monopterus albus
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Cell & Bioscience Open Access
RESEARCH
DNA methylation modification is associated with gonadal differentiation in Monopterus albus Xin Wang1, Fengling Lai1, Jun Xiong2, Wang Zhu1, Bifeng Yuan2, Hanhua Cheng1* and Rongjia Zhou1*
Abstract Background: Both testis and ovary can be produced sequentially in an individual with the same genome when sex reversal occurs in the teleost Monopterus albus, and epigenetic modification is supposed to be involved in gonadal differentiation. However, DNA methylation regulation mechanism underlying the gonadal differentiation remains unclear. Results: Here, we used liquid chromatography-electrospray ionization tandem mass spectrometry (LC–ESI–MS/MS) to simultaneously determine endogenous levels of both 5-methyl-2′-deoxycytidine (m5dC) and 5-hydroxymethyl2′-deoxycytidine (hm5dC) during gonadal differentiation. Overall DNA methylation level was upregulated from ovary to testis via ovotestis. As a de novo methylase, dnmt3aa expression was also upregulated in the process. Notably, we determined transcription factor Foxa1 for dnmt3aa gene expression. Site-specific mutations and chromatin immunoprecipitation showed that Foxa1 can bind to and activate the dnmt3aa promoter. Furthermore, DNA methylation levels of key genes foxl2 (forkhead box L2) and cyp19a1a (cytochrome P450, family 19, subfamily A, polypeptide 1a) in regulation of female hormone synthesis were consistently upregulated during gonadal differentiation. Conclusions: These data suggested that dynamic change of DNA methylation modification is associated with gonadal differentiation. Keywords: DNA methylation, Gonad, Promoter, Vertebrates Introduction DNA cytosine methylation (DNA methylation) is a major epigenetic modification that plays vital roles in animal development. During germ cell development, epigenetic reprogramming occurred dynamically, remodeling of DNA methylation marks in particular [1]. At day 7.5 of early embryo, global DNA methylation of primordial germ cells (PGCs) were erased and new DNA methylation were then established during late stage of development in mouse [2]. However, de novo DNA methylation *Correspondence: [email protected]; [email protected] 1 Hubei Key Laboratory of Cell Homeostasis, College of Life Sciences, Wuhan University, Wuhan 430072, China Full list of author information is available at the end of the article
proceeded differentially between male and female germ cells, earlier in spermatogenesis than in oogenesis. In the female germline cells, de novo DNA methylation occurred in arrested oocytes in meiotic prophase I. However, in the male germline cells, it took place in mitotically arrested prospermatogonia before birth [3, 4]. Reprogramming of DNA methylation not only occurs during germ cell differentiation, but also after fertilization. DNA methylation marks in sperms were quickly erased in the zygote by active demethylation mechanism [5], in contrast, the methylation marks in oocytes were lost via passive dilution through cell division [1]. The DNA methylation was established b
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