哺乳动物早期胚胎的基因表达及其调控

哺乳动物的早期发育包括合子的形成、胚胎基因组的活化和细胞开始分化等。在这段时期,精蛋白被组蛋白取代;二倍体形成后甲基化的单倍体亲本基因组经历了脱甲基作用;母本控制的发育被合子控制所取代。此外,在染色体调节的转录抑制状态形成之后,胚胎基因组活化,但基因的有效表达需要有增强子。这种转录抑制状态的产生很可能发生在染色质结构水平,因为诱导组蛋白过乙酰化可免除对增强子的需求。早期胚胎的mRNA表达模式与在体内或体外成功发育的关系,对确定最佳培养条件和核移植程序是必不可少的。     

小鼠胚胎ZGA中的基因表达及调控机制

哺乳动物早期胚胎发育的关键事件之一就是合子型基因激活,它决定着胚胎的正常发育与否。早期ZGA的起始必须依赖于源自母亲的母型蛋白,而这些蛋白的翻译后修饰过程,尤其是蛋白质磷酸化过程在ZGA中起重要作用。在小鼠中,伴随2细胞胚胎的形成,ZGA发生时选择性激活基因往往需要增强子的作用。由于合子钟以及2细胞胚胎期的转录抑制环境的影响,避免了那些应该在胚胎发育后期表达的基因的成熟前表达,使它们能按正确的发育时空顺序被选择性激活。目前,已经发现了在ZGA初期被激活的一些合子型基因,诸如hsp70、eIF-4C、U2afbp-rs等,对它们的研究已有了新的进展。     

组蛋白在胚胎早期发育转录调控中的作用

组蛋白是一组等电点大于１０．０的碱性蛋白质,在进化上十分保守。真核生物的染色体上主要含有５种组蛋白,即核心组蛋白Ｈ２ａ、Ｈ２ｂ、Ｈ３、Ｈ４及连接组蛋白Ｈ１。核心组蛋白八聚体、蛋白Ｈ１和２００ｂｐ的ＤＮＡ共同组成了染色体的基本单位——核小体。组蛋白不仅...     

小鼠早期胚胎发育过程中细胞凋亡及凋亡基因表达的检测

小鼠早期胚胎发育过程中凋亡现象大量存在,细胞凋亡与凋亡基因表达有关。应用彗星电泳法检测小鼠早期胚胎凋亡情况;应用巢式RT-PCR、免疫组化的方法检测了Bcl-2家族成员(Bax、Bcl-2、Bak、Bcl-xl)的表达变化情况。结果显示:随着胚胎细胞数目的增加,凋亡比率逐渐增大;Bax表达量在整个过程中基本不变,Bcl-2表达量逐渐上调,Bak、Bcl-xl的表达量逐渐降低。对小鼠早期胚胎发育过程中的基因表达研究对于揭示早期胚胎发育的机制有重大的意义。     

植物胚胎发生过程的基因表达

植物胚胎发生过程的基因表达黄绍兴,阎隆飞（北京农业大学生物学院,北京１０００９４）ＧＥＮＥＥＸＰＲＥＳＳＩＯＮＤＵＲＩＮＧＰＬＡＮＴＥＭＢＲＹＯＧＥＮＥＳＩＳ￥ＨｕａｎｇＳｈａｏ－ｘｉｎｇ;ＹａｎＬｏｎｇ－ｆｅｉ（ＣｏｌｌｅｓｅｏｆＢｉｏｌｏｇｉｃａ...     

多组学大数据整合分析揭示早期胚胎发育过程中基因表达调控信息的变化规律

摘要 目的：探究哺乳动物早期胚胎发育过程中基因表达调控信息的变化规律。方法：收集小鼠早期胚胎发育各时期的RNA-seq,ATAC-seq,MethylC-Seq和H3K4me3 ChIP-seq数据进行整合分析,观察小鼠早期胚胎发育各时期转录因子表达量的变化,计算各时期基因表达量与转录因子结合位点数量及染色质可及性的相关性,筛选各时期表达量前10%的基因,统计其表达量和转录因子占比,并进行启动子可及性分析。根据前期报道的转录因子三节点调控网络,对早期胚胎各时期转录因子调控网络的富集模式进行分析。根据多组学数据分析结果,推测早期胚胎发育调控过程中转录因子和表观遗传修饰信息的共调控模型。结果：转录因子数量和调控关系变化以及染色质可及性、DNA甲基化修饰、组蛋白修饰等表观遗传修饰共同调控早期胚胎发育各时期的基因表达,这些因素在不同时期发挥不同程度的调控作用。结论：转录因子和表观遗传修饰在早期胚胎发育过程中动态调控基因表达。     

胚胎植入的调控机理

胚胎植入对于妊娠的建立和维持至关重要,需要活化的胚泡和接受态的子宫之间进行同步。在辅助生殖技术中,子宫接受态的判断仍是制约妊娠率的一个关键限制性因素。已有数据显示,胚胎植入涉及一系列信号分子的激活和失活,进而影响子宫腔上皮细胞的增殖与分化、上皮极性、宫腔闭合、胚胎定位、上皮基质反应、腺体发育等。本文就雌激素、孕酮、白血病抑制因子(leukemia inhibitory factor, LIF)、microRNA (miRNA)、通道蛋白、信号转导通路等在胚胎植入过程中的作用及其调控网络作一综述,以期为不孕症的治疗及安全有效的避孕药开发提供理论依据。     

哺乳动物卵母细胞及早期胚胎核仁结构与rRNA基因表达研究进展

核仁是细胞中合成哺rRNA的场所,与细胞的蛋白质合成密切相关,近年的研究证明,在卵子发生和早期胚胎发育过程中,核仁的结构和机能都发生着有规律的变化,对于研究卵子发生和早期胚胎发育机理有着重要的意义。本文以牛为主要对象,综述了有关核仁的结构,发生以及卵母细胞成熟及早期胚胎发育过程中核仁结构和rRNA转录变化规律的研究进展。     

昆虫基因表达的调控

昆虫的基因表达过程十分复杂,受多种调控因子的调节,随着分子生物学研究新技术在昆虫学上的应用,昆虫基因表达的调控也取得了较快进展,本文以一些特征清楚的系统为例,讨论昆虫基因表达调控的研究现状,旨在为其它昆虫表达的调控研究提供借鉴。     

Tet-on基因表达系统转染胚胎干细胞和调控效应检测*

以荧光素酶基因为目的基因, 应用Lipofectamine介导的瞬时转染方法, 将pTet-on调控质粒和pTRE2hyg-luciferase表达质粒共转染小鼠胚胎干细胞, 滴加系列浓度的DOX后, 观察其诱导效应. 结果显示, 随DOX浓度升高, 荧光素酶的活性也逐渐增强; 而未转染细胞与转染细胞未加诱导剂时, 其RLU值均接近空白对照. 表明转染Tet-on基因表达系统的胚胎干细胞对DOX诱导有较好的反应性, 目的基因的表达与DOX具有严格的剂量依赖关系.     

Characterization of gene expression in double-muscled and normal-muscled bovine embryos

Myostatin, a member of the transforming growth factor-beta superfamily, is a negative regulator of skeletal muscle growth. Cattle with mutations that inactivate myostatin exhibit a remarkable increase in mass of skeletal muscle called double muscling that is accompanied by an equally remarkable decrease in carcass fat. Although a mouse knockout model has been created which results in mice with a 200% increase in skeletal muscle mass, molecular mechanisms whereby myostatin regulates skeletal muscle and fat mass are not fully understood. Using suppressive subtractive hybridization, genes that were differentially expressed in double-muscled vs. normal-muscled cattle embryos were identified. Genetic variation at other loci was minimized by using embryonic samples collected from related Piedmontese x Angus dams or Belgian Blue x Hereford dams bred to a single sire of the same breed composition. Embryos were collected at 31-33 days of gestation, which is 2-4 days after high-level expression of myostatin in the developing bovine embryo. The suppressive subtraction resulted in 30 clones that were potentially differentially expressed, 19 of which were confirmed by macroarray analysis. Several of these genes have biological functions that suggest that they are directly involved in myostatin's regulation of skeletal muscle development. Furthermore, several of these genes map to quantitative trait loci known to interact with variation in the myostatin gene.     

Evaluation of light regulatory potential of Calvin cycle steps based on large-scale gene expression profiling data

Although large-scale gene expression data have been studied from many perspectives, they have not been systematically integrated to infer the regulatory potentials of individual genes in specific pathways. Here we report the analysis of expression patterns of genes in the Calvin cycle from 95 Arabidopsis microarray experiments, which revealed a consistent gene regulation pattern in most experiments. This identified pattern, likely due to gene regulation by light rather than feedback regulations of the metabolite fluxes in the Calvin cycle, is remarkably consistent with the rate-limiting roles of the enzymes encoded by these genes reported from both experimental and modeling approaches. Therefore, the regulatory potential of the genes in a pathway may be inferred from their expression patterns. Furthermore, gene expression analysis in the context of a known pathway helps to categorize various biological perturbations that would not be recognized with the prevailing methods.     

Global gene expression analysis of bovine blastocysts produced by multiple methods

Reproductive efficiency using somatic cell nuclear transfer (SCNT) technology remains suboptimal. Of the various efforts to improve the efficiency, chromatin transfer (CT) and clone-clone aggregation (NTagg) have been reported to produce live cloned animals. To better understand the molecular mechanisms of somatic cell reprogramming during SCNT and assess the various SCNT methods on the molecular level, we performed gene expression analysis on bovine blastocysts produced via standard nuclear transfer (NT), CT, NTagg, in vitro fertilization (IVF), and artificial insemination (AI), as well as on somatic donor cells, using bovine genome arrays. The expression profiles of SCNT (NT, CT, NTagg) embryos were compared with IVF and AI embryos as well as donor cells. NT and CT embryos have indistinguishable gene expression patterns. In comparison to IVF or AI embryos, the number of differentially expressed genes in NTagg embryos is significantly higher than in NT and CT embryos. Genes that were differentially expressed between all the SCNT embryos and IVF or AI embryos are identified. Compared to AI embryos, more than half of the genes found deregulated between SCNT and AI embryos appear to be the result of in vitro culture alone. The results indicate that although SCNT methods have altered differentiated somatic nuclei gene expression to more closely resemble that of embryonic nuclei, combination of insufficient reprogramming and in vitro culture condition compromise the developmental potential of SCNT embryos. This is the first set of comprehensive data for analyzing the molecular impact of various nuclear transfer methods on bovine pre-implantation embryos.