早期胚胎细胞谱系研究——揭示生命发育早期的黑盒子

哺乳动物的早期胚胎发育,通过多个层次的细胞命运决定,建立了胚胎器官发生、形态建成的整体发育蓝图,是生命体最重要的分子事件之一。早期胚胎发育过程伴随了全能性的维持和分化,以及各种多能干细胞命运的次序决定,任何发育进程上的缺陷都会对整个胚胎个体产生深远的影响。因此,研究早期胚胎谱系建立的过程、不同胚层和组织前体细胞的命运决定及其发生与发展的调控机制,不仅仅是面对国家人口政策的变化以及优生优育的需求,预防和减少早期发育疾病,还能够指导胚胎干细胞及各种多能干细胞的分化和进一步转化医学应用,因而具有极其重要的生物学意义。     

胚胎干细胞向内胚层分化的分子机制及其组学研究进展

胚胎干细胞(ESC)在发育过程中分化为内胚层、中胚层和外胚层3个胚层.其中内胚层进一步向终末细胞的分化,是形成整个消化道和呼吸道,以及肝脏、胰腺等器官的基础.ESC形成内胚层主要经历以下几个分化阶段:外胚叶的分化、原条的形成、内胚层与中胚层的分离以及定型内胚层的形成.本文主要从信号通路、转录因子以及表观遗传调控等几个方面综述胚胎干细胞向内胚层分化的分子机制,并重点介绍其组学研究进展,以期为该领域研究者提供重要参考信息.     

简述两栖类的原肠作用及三胚层形成

在讲到动物发育时,常遇到一个不易理解和讲述的动态过程,就是蛙胚的原肠发生过程及三胚层的形成。原肠胚是胚胎发育中一个极为重要的时期,胚胎从囊胚未分化的分裂球发育成为具有内、中、外三个胚层的原肠胚,并将决定其各器官原基,它是通过空间和时间方面的紧密结合,复杂的形态发生运动而完成的。内胚层形成消化道     

利用胚胎干细胞分化的拟胚体研究小鼠早期胚胎发育过程

小鼠胚胎干细胞(ES细胞)具有分化的全能性已经得到广泛共识。ES细胞在体外分化所形成的拟胚体在结构上能够模仿早期胚胎发育过程,包括在内细胞团表面形成内胚层、柱状上皮细胞的分化,以及中央空腔的形成。本文介绍利用拟胚体研究小鼠早期胚胎发育过程中各个胚胎阶段的发育、细胞程序性死亡的发生及TGF-β信号在胚胎发育过程中的作用。     

斑马鱼胚胎背腹轴建立的分子机制

脊椎动物胚胎发育起始于体轴的建立,是胚胎早期发育过程中最重要的事件之一。Wnt、BMP、Nodal和FGF等多个信号通路协同调控细胞分化和细胞运动,促进胚胎胚层的形成和空间上的分离,调控胚胎背腹轴、前后轴和左右轴线的分化,为胚胎进一步发育勾勒出蓝图。本文主要综述斑马鱼胚胎背腹轴建立的分子机制,包括背部组织中心简介;母源Wnt/β-catenin信号调控背部组织中心形成的分子机制;BMP信号调控背腹轴建立的分子机制。     

胚胎干细胞向中内胚层分化的调控机制

在哺乳动物胚胎发育过程中,中内胚层(mesendoderm)也被称为原条(primitive streak),是中胚层和内胚层分化的过渡时期。中内胚层的存在时间较短,但成功的中内胚层分化对随后进行的中胚层与内胚层发育至关重要。发育生物学的研究极大地推动了人们对胚胎发育的认识,同时,越来越多体外分化系统的建立也加深了对环境信号如何影响胚层分化的理解。近些年来,通过表观遗传的研究,人们逐渐认识到染色体结构与组蛋白修饰的改变也在分化发育过程中起到重要作用。通过胚胎干细胞定向诱导中内胚层分化来探究相关分子机制,不仅有助于对早期胚胎发育的了解,也有助于临床应用与疾病治疗。现总结了TGF-β信号、Wnt信号和FGF信号调控中内胚层分化的研究现状,并概述了这些信号如何与表观修饰共同调控胚胎干细胞向中内胚层分化的进展。     

小鼠胚胎干细胞分化形成拟胚体过程中的细胞程序性死亡

为了检测小鼠胚胎干细胞 (embryonicstemcell ,ES细胞 )体外分化的拟胚体 (embryoidbodies ,EBs)形成过程中细胞程序性死亡 (programmedcelldeath ,PCD)的发生 ,通过悬滴、悬浮培养技术定向诱导未分化的ES细胞分化为拟胚体 ,并用RT PCR检测原始内胚层、原始外胚层、中胚层、内脏内胚层 4种分子标记物在EBs中的表达 .通过TUNEL染色、电镜、激光共聚焦显微镜及Western印迹以确定凋亡发生 .结果表明 :ES细胞体外分化为拟胚体并且表达各胚层相应的分子标记物 ;在拟胚体的发育过程中出现明显的空腔化过程 ,TUNEL染色及电镜观察到凋亡生成 ,同时线粒体膜电位 (ΔΨm)在拟胚体发育过程中降低 ,通过Western印迹检测到caspase3、caspase8的激活 .表明小鼠ES细胞所分化的拟胚体可以作为研究早期胚胎发育的实验模型 ,线粒体在拟胚体的细胞程序性死亡过程中发挥重要的作用 .为进一步利用拟胚体研究细胞程序性死亡及相关信号分子在小鼠胚胎发育早期的作用奠定了基础     

家兔胚泡ICM在体外培养系统中的行为

一、用显微外科方法从兔早期胚泡分离来的ICM在所用的体外培养系统中的行为,因ICM的生长状态而不同。呈带状生长的ICM从近端向远处延伸,而细胞的分化则从远端逐渐向近端进行,它具有明显的极性。细胞的分化秩序井然,层次分明。因此呈带状生长的ICM适用于进行细胞分化和细胞谱系的研究。二、呈团块状生长的ICM,没有明显的极性,细胞分化开始于团块的外表面,逐渐向中央进行。细胞分化开始稍晚一些,速度也较慢,这种生长状态的ICM适用于分离胚胎干细胞。三、兔子胚外内胚层从ICM分化来,它经历了二次分化的过程。第一次发生于培养后第三天(相当于交配后第七天),形成第一胚外内胚层-胚外体壁内胚层,它向远处迁移,远离原始外胚层。培养后四天(相当于交配后第八天),形成第二种胚外内胚层-胚外脏壁内胚层,它尾随体壁内胚层向远处迁移,其后缘的部分细胞向原始外胚层附近的滋胚层下面侵润。四、胚外体壁内胚层和脏壁内胚层的细胞是由少数决定了的和离开ICM的细胞分化和增殖而来。五、文中讨论了早期胚胎细胞分化的谱系问题。     

胚胎性干细胞定向诱导分化——新崛起的细胞工程

胚胎性干细胞（ＥＳ）细胞）是一类稳定地在体外自我增殖、具有发育全能性和产生属于三个胚层范畴分化细胞、甚至参与体内个体完整发育的早期胚胎细胞。包括人类在内的各种哺乳类ＥＳ细胞分离建系成功,结合体外基因操作和细胞分化诱导条件选择等综合途径,有可能控制ＥＳ细胞导向产生单一类型的功能性分化细胞。这种全能ＥＳ细胞生物学技术已成为一项崛起的细胞工程。在新世纪１０－１５年内将对分子细胞生物学、发育生物学和人体胚     

Nodal信号的研究进展

早期胚胎发育过程中,组织者能够诱导胚胎组织形成完整的体轴。研究表明细胞之间的信息交流是脊椎动物胚胎诱导作用的基础。Nodal作为早期胚胎诱导信号的关键成分,参与了中胚层和内胚层的形成、前-后体轴的位置确定和左-右体轴特化等一系列关键事件,表明Nodal信号在脊椎动物早期发育过程中具有重要作用。     

Endoderm development in vertebrates: fate mapping, induction and regional specification

The formation of the vertebrate body plan begins with the differentiation of cells into three germ layers: ectoderm, mesoderm and endoderm. Cells in the endoderm give rise to the epithelial lining of the digestive tract, associated glands and respiratory system. One of the fundamental problems in developmental biology is to elucidate how these three primary germ layers are established from the homologous population of cells in the early blastomere. To address this question, ectoderm and mesoderm development have been extensively analyzed, but study of endoderm development has only begun relatively recently. In this review, we focus on the 'where', 'when' and 'how' of endoderm development in four vertebrate model organisms: the zebrafish, Xenopus, chick and mouse. We discuss the classical fate mapping of the endoderm and the more recent progress in characterizing its induction, segregation and regional specification.     

Early endoderm development in vertebrates: lineage differentiation and morphogenetic function

Gastrulation of the vertebrate embryo culminates in the formation of three primary germ layers: ectoderm, mesoderm and endoderm. The endoderm contributes to the lining of the gut and the associated organs. New components of the molecular pathway for endoderm specification have been identified in the zebrafish and Xenopus. In the mouse, the activity of orthologous factors is involved with the allocation and differentiation of the definitive endoderm. Morphogenetic interactions between the endoderm and the other germ layer derivatives are critical for the morphogenesis of head structures and organogenesis of gut derivatives.     

Expression of the cell surface-associated glycoprotein, fibronectin, in the early mouse embryo.

The expression of the cell surface-associated glycoprotein fibronectin was studied by indirect immunofluorescence in the early stages of mouse embryogenesis. Fibronectin was not detectable in early preimplantation embryos. Trace amounts of the protein were first found between the cells of the inner cell mass of late blastocysts. In implanted early egg cylinders, fibronectin was deposited between the ectoderm and endoderm of the inner cell mass and in the nascent Reichert's membrane. With development, the visceral and the parietal endoderm cells became positive for the protein, but no fibronectin was detected in ectoderm cells. During segregation of mesoderm from ectoderm, fibronectin appeared in mesoderm cells and as a band between the two germ layers. In the developing amnion and chorion, the protein was localized between the ectodermal and mesodermal cell layers. The results indicate that fibronectin is an early differentiation market for the stage of endoderm formation in the inner cell mass of the mouse blastocyst. It is also a marker of mesoderm appearance and seems to be associated with the accumulating extracellular matrix material in the developing embryo.     

Definition of germ layer cell lineage alternative splicing programs reveals a critical role for Quaking in specifying cardiac cell fate

Alternative splicing is critical for development; however, its role in the specification of the three embryonic germ layers is poorly understood. By performing RNA-Seq on human embryonic stem cells (hESCs) and derived definitive endoderm, cardiac mesoderm, and ectoderm cell lineages, we detect distinct alternative splicing programs associated with each lineage. The most prominent splicing program differences are observed between definitive endoderm and cardiac mesoderm. Integrative multi-omics analyses link each program with lineage-enriched RNA binding protein regulators, and further suggest a widespread role for Quaking (QKI) in the specification of cardiac mesoderm. Remarkably, knockout of QKI disrupts the cardiac mesoderm-associated alternative splicing program and formation of myocytes. These changes arise in part through reduced expression of BIN1 splice variants linked to cardiac development. Mechanistically, we find that QKI represses inclusion of exon 7 in BIN1 pre-mRNA via an exonic ACUAA motif, and this is concomitant with intron removal and cleavage from chromatin. Collectively, our results uncover alternative splicing programs associated with the three germ lineages and demonstrate an important role for QKI in the formation of cardiac mesoderm.     

Early mouse endoderm is patterned by soluble factors from adjacent germ layers

Endoderm that forms the respiratory and digestive tracts is a sheet of approximately 500-1000 cells around the distal cup of an E7.5 mouse embryo. Within 2 days, endoderm folds into a primitive gut tube from which numerous organs will bud. To characterize the signals involved in the developmental specification of this early endoderm, we have employed an in vitro assay using germ layer explants and show that adjacent germ layers provide soluble, temporally specific signals that induce organ-specific gene expression in endoderm. Furthermore, we show that FGF4 expressed in primitive streak-mesoderm can induce the differentiation of endoderm in a concentration-dependent manner. We conclude that the differentiation of gastrulation-stage endoderm is directed by adjacent mesoderm and ectoderm, one of the earliest reported patterning events in formation of the vertebrate gut tube.     

The role of mesodermal signals during liver organogenesis in zebrafish

Three germ cell layers, the ectoderm, mesoderm and endoderm, are established during the gastrulation stage. All cell types in different organs and tissues are derived from these 3 germ cell layers at later stages. For example, skin epithelial cells and neuronal cells are derived from the ectoderm, while endothelial cells and muscle cells from the mesoderm and lung, and intestine epithelial cells from the endoderm. While in a normal situation different germ cells are destined to specific cell fates in differ...