The Role of DNA Methylation Reprogramming During Sex Determination and Transition in Zebrafish

DNA methylation is a prevalent epigenetic modification in vertebrates, and it has been shown to be involved the regulation of gene expression and embryo development. However, it remains unclear how DNA methylation regulates sexual development, especially in species without sex chromosomes. To determine this, we utilized zebrafish to investigate DNA methylation reprogramming during juvenile germ cell development and adult female-to-male sex transition.We reveal that primordial germ cells(PGCs) undergo significant DNA methylation reprogramming during germ cell development, and the methylome of PGCs is reset to an oocyte/ovary-like pattern at 9 days post fertilization(9 dpf). When DNA methyltransferase(DNMT) activity in juveniles was blocked after 9 dpf, the zebrafish developed into females. We also show that Tet3 is involved in PGC development. Notably, we find that DNA methylome reprogramming during adult zebrafish sex transition is similar to the reprogramming during the sex differentiation from 9 dpf PGCs to sperm. Furthermore, inhibiting DNMT activity can prevent the female-to-male sex transition, suggesting that methylation reprogramming is required for zebrafish sex transition. In summary, DNA methylation plays important roles in zebrafish germ cell development and sexual plasticity.     

Biochemical characterization of the placeholder nucleosome for DNA hypomethylation maintenance

DNA methylation functions as a prominent epigenetic mark, and its patterns are transmitted to the genomes of offspring. The nucleosome containing the histone H2A.Z variant and histone H3K4 mono-methylation acts as a “placeholder” nucleosome for DNA hypomethylation maintenance in zebrafish embryonic cells. However, the mechanism by which DNA methylation is deterred by the placeholder nucleosome is poorly understood. In the present study, we reconstituted the placeholder nucleosome containing histones H2A.Z and H3 with the Lys4 mono-methylation. The thermal stability assay revealed that the placeholder nucleosome is less stable than the canonical nucleosome. Nuclease susceptibility assays suggested that the nucleosomal DNA ends of the placeholder nucleosome are more accessible than those of the canonical nucleosome. These characteristics of the placeholder nucleosome are quite similar to those of the H2A.Z nucleosome without H3K4 methylation. Importantly, the linker histone H1, which is reportedly involved in the recruitment of DNA methyltransferases, efficiently binds to all of the placeholder, H2A.Z, and canonical nucleosomes. Therefore, the characteristics of the H2A.Z nucleosome are conserved in the placeholder nucleosome without synergistic effects on the H3K4 mono-methylation.     

早期胚胎发育合子基因组激活调控

动物早期胚胎发育始于分化成熟的雌雄配子经受精后重编程为全能性合子。在胚胎发育的初期,合子基因组的转录水平处于静默状态,母源物质调控占据主导地位。随着胚胎发育的进行,母源物质会经历分阶段的降解,合子基因组开始逐渐激活转录,标志着早期胚胎发育从母源性调控向合子基因组调控的转变,也称为母源-合子转换（maternal-zygotic transition,MZT）。其中一个关键的转折性事件就是合子基因组激活（zygotic genome activation,ZGA）,ZGA的正确发生对于早期胚胎发育和细胞命运决定至关重要。然而,目前对于ZGA的调控因子和具体的分子机制仍知之甚少。研究表明,ZGA在不同物种中存在较大差异,可能受到DNA甲基化、组蛋白修饰、非编码RNA、染色质重塑以及ZGA相关因子等多种调控因素的影响。本文探讨了上述几种调控因素影响合子基因组激活的研究进展,对进一步研究早期胚胎ZGA的相关机制具有借鉴意义。     

斑马鱼中囊胚过渡调控机制

斑马鱼中囊胚过渡(MBT)始于受精卵的第10次卵裂,此时亦伴有细胞周期延长,分裂同步性丧失,合子型基因开始转录活化,胚胎细胞开始具备运动迁移能力等现象。斑马鱼MBT。的发生依赖于胚胎细胞的核质比,胚胎细胞周期中的G1时相则只有在合子型基因组开始被转录活化后才能出现。细胞周期检验点的激活可能也是受转录调控的,但中期检验点对DNA复制抑制状态的响应不仅在MBT前后、甚至在MBT前的不同阶段也可能有具体作用途径的差异。活化的P38蛋白在胚胎中的不对称分布是维持卵裂阶段细胞分裂同步性的关键因素。尽管大规模的合子型基因的表达发生在MBT开始后,也有少数与胚层分化有关的合子型基因是在MBT。前表达的,还有一些既有母型表达也有合子型表达的基因在MBT前后分别参与不同的信号途径来调控胚胎的发育与分化。     

DNA Methylation and Epigenotypes

The science of epigenetics is the study of all those mechanisms that control the unfolding of the genetic program for development and determine the phenotypes of differentiated cells. The pattern of gene expression in each of these cells is called the epigenotype. The best known and most thoroughly studied epigenetic mechanism is DNA methylation, which provides a basis both for the switching of gene activities, and the maintenance of stable phenotypes. The human epigenome project is the determination of the pattern of DNA methylation in multiple cell types. Some methylation sites, such as those in repeated genetic elements, are likely to be the same in all cell types, but genes with specialized functions will have distinct patterns of DNA methylation. Another project for the future is the study of the reprogramming of the genome in gametogenesis and early development. Much is already known about the de novo methylation of tumor suppressor genes in cancer cells, but the significance of epigenetic defects during ageing and in some familial diseases remains to be determined.     

DNA甲基化与动脉粥样硬化

DNA甲基化是一种重要的表观遗传调控方式,可在转录前水平调节基因的表达.近年来的研究表明,动脉粥样硬化的发生发展与DNA甲基化密切相关. 对DNA甲基化模式改变在动脉粥样硬化发病的相关机制做深入研究,可能为动脉粥样硬化的诊治提供一种新的途径.本文将从基因组低甲基化、相关基因异常甲基化以及动脉粥样硬化危险因素的DNA甲基化等方面重点阐述DNA甲基化与动脉粥样硬化的关系.     

Methylation of mitochondrial DNA in carrot (Daucus carota L.)

The methylation status of carrot (Daucus carota L.) mitochondrial DNA (mtDNA) was studied using isoschizomeric restriction enzymes MspI/HpaII (CCGG) and MvaI/EcoRII [CC(A/T)GG]. Southern hybridisations with probes for mitochondrial genes coxII and atpA were performed. MtDNAs isolated from non-embryogenic cell suspensions and roots were analysed. No differences were found using MspI/HpaII but after digesting the mtDNA with MvaI and EcoRII, some qualitative and quantitative differences between the restriction patterns appeared. Distinction was also revealed after Southern hybridisation with the coxII probe. These data indicate that the mtDNA of carrot is methylated in CNG trinucleotides and unmethylated in CG dinucleotides in CCGG sequences. The results were reproducible for cell suspensions of various genotypes and even cultivars but the extent of methylation was different in the root. The possible role of methylation in the mitochondrial genome of higher plants is discussed. Received: 16 April 1997 / Revision received: 4 July 1997 / Accepted: 30 July 1997     

植物DNA甲基化

DNA甲基化是造成植物转录水平基因沉默的主要原因。从DNA甲基化的发生机理,DNA甲基化抑制基因转录以及调控基因转录的方式简要地介绍了真核生物中DNA甲基化的功能和调控机制方面的一些研究进展。     

植物DNA甲基化及其表观遗传作用

表观遗传学(epigenetics)是研究没有DNA序列变化的、可遗传的基因表达的改变。目前研究表明,表观遗传学在植物生长发育过程中起着极其重要的作用,主要通过包括DNA甲基化、RNA干涉、基因组印记、转基因沉默等多个方面来调控植物的生长发育。其中,DNA甲基化是表观遗传学的最重要研究内容之一,是调节基因组功能的重要手段。现对植物DNA甲基化的特征、维持机制、调控机制、表观遗传作用及其研究方法进行简要论述。     

DNA甲基化在动植物遗传育种中的研究进展

DNA甲基化是真核生物表观遗传学重要的机制之一,对基因转录水平的表达具有重要的调控作用。近年来,DNA甲基化在动植物遗传育种领域的研究引起了人们广泛的关注。我们从DNA甲基化与基因的表达调控、动植物基因组的甲基化状态、甲基敏感扩增片段多态性方法、DNA甲基化与杂种优势,以及DNA甲基化与分子标记等方面,简要综述了国内外有关DNA甲基化在动植物遗传育种研究中的进展,着重于全基因组DNA甲基化模式在动植物遗传育种中的相关研究和应用。     

DNA甲基化与microRNA

细胞中DNA甲基化和microRNA（miRNA）相互影响,并共同调控着下游靶基因的表达活性,在细胞生长代谢、免疫、肿瘤和心血管疾病等生理和病理过程中发挥重要作用。首先简要介绍DNA甲基化与miRNA的概况,然后分析了miRNA调控下的DNA甲基化改变,探讨了DNA甲基化影响miRNA的表达活性变化,并归纳了miRNA与DNA甲基化之间的反馈调控关系;最后对DNA甲基化和miRNA的应用前景进行了简单探讨。研究DNA甲基化与miRNA间的网络调控关系,可为表观调控机制在理论和实践中的深入研究和应用提供参考。     

DNA甲基化与癌症

DNA甲基化是重要的表观遗传修饰,主要发生在DNA的CpG岛. DNA的甲基化通过DNA甲基转移酶（DNA methyltransferases, DNMTs）完成. DNA甲基化参与了细胞分化、基因组稳定性、X染色体失活、基因印记等多种细胞生物学过程.单基因水平及基因组范围内的DNA甲基化改变在肿瘤发生发展中亦发挥重要作用. 抑癌基因的异常甲基化引起的表达抑制,可导致肿瘤细胞的增殖失控和侵袭转移,并参与肿瘤组织的血管生成过程.在许多肿瘤的研究中都发现了基因组整体DNA低甲基化所导致的染色体不稳定性. 本文从DNA的异常高甲基化和低甲基化两方面论述了DNA甲基化在细胞恶变发生发展过程中的改变及其影响,并阐述了DNA甲基化改变在肿瘤诊断和治疗中的作用.     

植物中DNA甲基化模式及其相关机制

DNA甲基化是表观遗传修饰的重要形式之一,是植物中较早发现的DNA共价修饰方式。在植物的正常生长发育中,DNA甲基化与植物基因组维持、体细胞无性系变异、外来基因防御、内源基因的表达、转基因沉默以及基因印迹之间有着极大的关系,因此,植物DNA甲基化的研究对植物基因工程的发展有着举足轻重的作用。本文介绍了参与DNA甲基化的各种酶和蛋白质,阐述了DNA甲基化相关机制的最新研究进展。