LIN28调控哺乳动物干细胞和生殖细胞发育分化的研究概述

Lin28为一种保守的RNA结合蛋白质,在细胞代谢、细胞周期和多能性维持中具有重要调控作用。近年来,发现Lin28在哺乳动物原始生殖细胞形成和分化、精原干细胞的形成、自我更新和分化调控中具有不可替代的作用。本文对Lin28作用及其对哺乳动物干细胞和生殖细胞发育分化影响的研究进展作一简述。     

综述: DNA甲基化在动物胚胎和生殖细胞发育过程中的重编程

表观遗传信息DNA甲基化在动物的发育、细胞分化和器官形成过程中,起着至关重要的作用．近期,关于DNA甲基化在脊椎动物胚胎发育和生殖细胞发育过程重编程的研究取得了重要的进展．发现斑马鱼的早期胚胎完整地继承了精子的DNA甲基化图谱,而哺乳动物的早期胚胎和原始生殖细胞发育过程则经历了整体去甲基化并重新建立甲基化图谱的过程,但胚胎发育过程中基因的印迹区未发生DNA去甲基化,而生殖细胞发育过程中印迹区的甲基化修饰被消除．     

DNA甲基化在动物胚胎和生殖细胞发育过程中的重编程

表观遗传信息DNA甲基化在动物的发育、细胞分化和器官形成过程中,起着至关重要的作用.近期,关于DNA甲基化在脊椎动物胚胎发育和生殖细胞发育过程重编程的研究取得了重要的进展.发现斑马鱼的早期胚胎完整地继承了精子的DNA甲基化图谱,而哺乳动物的早期胚胎和原始生殖细胞发育过程则经历了整体去甲基化并重新建立甲基化图谱的过程,但胚胎发育过程中基因的印迹区未发生DNA去甲基化,而生殖细胞发育过程中印迹区的甲基化修饰被消除.     

原始生殖细胞发生的机制

生殖细胞的发生是发育和遗传的基础。在几乎所有哺乳动物中,原始生殖细胞（primordial germ cell,PGC）均由近端上胚层体细胞在周边细胞特定的信号诱导下特化而成。目前的研究已经发现一些与生殖细胞特化有关的信号分子和关键转录调控元件,以及特化后生殖细胞获得的与体细胞不同的生物特性。生殖细胞的特化是一个结合了体细胞发育程序的抑制、细胞多能性程序的启动和全基因组表观遗传重编程三个方面的动态的复杂过程。多能性干细胞（胚胎干细胞或诱导型多能干细胞）具有发育全能性,能分化为机体任何一种细胞类型,包括生殖细胞。利用多能性干细胞体外分化形成生殖细胞有助于深入系统地研究配子发生的调控机制,为干细胞在不育症治疗方面的应用带来新希望。     

名刊封面

《自然》2005.7.14与胚胎发育Blimpl(一种甲基转移酶)是淋巴液中 B-细胞分化的"总调节因子",而 B-细胞分化是成人免疫系统的重要组成部分。令人吃惊的是,研究人员现在发现,Blimp 1在原始生殖细胞的指定过程中也扮演着关键角色。最初的原始生殖细胞从将形成身体其余部分的相邻细胞中隔离出来,最终进入发育胚胎的生殖     

TET蛋白质在动物原始生殖细胞发育过程中的作用

原始生殖细胞(primordial germ cell,PGC)是指能发育成精子或卵子的祖先细胞。PGC在向生殖细胞发育过程中会经历原有基因印记的去除、新印记的形成和维持,而DNA去甲基化是维持这一过程的主要机制。研究发现,TETs(ten-eleven translocations)蛋白质可以通过不同机制调控DNA去甲基化过程,在原始生殖细胞的形成和胚胎发育过程中发挥重要作用。该文综述了TET家族的结构、调控去甲基化的机制及其在动物原始生殖细胞发育过程中的作用。     

SOX转录因子家族在生殖细胞命运决定中的调控作用

SOX(SRY-related HMG-box)是一类含有HMG box DNA结合结构域的转录因子,自第一个SOX成员—性别决定基因SRY发现以来,先后共发现了20个成员。SOX不仅在胚胎发育、组织自稳态、神经发育、视网膜发育、骨组织形成等方面发挥重要调控功能,而且在生殖细胞的发生、分化和成熟等过程中也有重要的调控作用,如SOX17能调控胚胎干细胞向生殖细胞分化的过程中,使胚胎干细胞在体外可以诱导形成生殖细胞,为解决人类的不育问题带来了曙光。该文对近年来SOX家族在生殖细胞发育和命运决定方面的研究进展情况作一简述。     

胚胎干细胞向造血细胞分化研究

胚胎干（embryonic stem,ES）细胞是来源于囊胚的内细胞团（inner cell mass,ICM）,具有发育的全能性或多能性,能嵌合到早期胚胎,在体内可以参与各种组织发育甚至包括生殖细胞;在体外分化培养条件下,可以顺序分化出各种组织细胞,与体内完整胚胎发育过程相符合,而且可以通过调节ES细胞某些基因的表达而调节其分化。因此,ES细胞是研究哺乳动物早期胚胎发育、细胞分化及其关键基因鉴定的理想模型。另外,胚胎生殖脊（embryonic germ,EG）细胞系也具有同样的生物学特性,它是由早期胚胎的原始生殖脊（primordial germ,PG）细胞建株而来。最近研究显示：ES细胞在体外不但可以分化为所有造血细胞系,而且还可以分化为具有长期增殖能力的造血干细胞。作者就胚胎干细胞向造血细胞和造血干细胞分化及其诱导因子和调控基因的表达作一综述。     

BMP-1及BMP家族在背-腹轴图式形成中的作用

骨形态发生蛋白（bone morphogenetic proteins, BMPs）是一类在发育过程中起重要作用的分子。除BMP-1外,其他BMP分子均属于转化生长因子-β（transforming growth factor-β, TGF-β）/BMP超家族的发育信号分子。在胚胎发育过程中,这些信号分子通过形成浓度梯度对背—腹轴各向异性分化进行调控。它们借助细胞表面受体的识别进行信号传导,参与调控细胞分化、增殖等活动。而BMP-1则属于细胞外基质金属蛋白酶超家族中的Tolloid蛋白酶家族。BMP-1通过水解其他BMP的抑制物（如脊索发生素,Chordin）,达到促进其他BMP信号传导的目的。BMP-1、BMP和Chordin三者通过相互制约与相互促进等一系列作用,在背—腹沿线建立起稳定的BMP信号梯度。本文就BMP浓度梯度的形成及其稳态维持的机制进行回顾与总结。并在此基础上,对各个物种间BMP浓度梯度形成机制的异同,以及可能存在的协同进化进行比较、分析和讨论。     

原始生殖细胞发生的机制

生殖细胞的发生是发育和遗传的基础。在几乎所有哺乳动物中,原始生殖细胞(primordial germ cell,PGC)均由近端上胚层体细胞在周边细胞特定的信号诱导下特化而成。目前的研究已经发现一些与生殖细胞特化有关的信号分子和关键转录调控元件,以及特化后生殖细胞获得的与体细胞不同的生物特性。生殖细胞的特化是一个结合了体细胞发育程序的抑制、细胞多能性程序的启动和全基因组表观遗传重编程三个方面的动态的复杂过程。多能性干细胞(胚胎干细胞或诱导型多能干细胞)具有发育全能性,能分化为机体任何一种细胞类型,包括生殖细胞。利用多能性干细胞体外分化形成生殖细胞有助于深入系统地研究配子发生的调控机制,为干细胞在不育症治疗方面的应用带来新希望。     

生殖细胞形成的调控机制及体外培养体系的研究进展

生殖细胞是多细胞生物体遗传物质传递的载体,在发育生物学、临床医学及畜牧业生产等领域中具有广阔的应用前景。原始生殖细胞作为胚胎体内最早出现的生殖细胞,在发育过程中受多种信号因子的诱导,发生特化、迁移、分化及减数分裂,最终形成单倍体的配子,此过程在遗传学和表观遗传学方面受到严格的调控。另外,多能性干细胞向生殖细胞的分化以及生殖细胞的体外培养方面在最近均取得了较大的进展。该文将主要围绕原始生殖细胞,综述最近几年来关于生殖细胞形成中的转录调控及体外培养体系的进展。     

Cell type-autonomous and non-autonomous requirements for Dmrt1 in postnatal testis differentiation

Genes containing the DM domain, a conserved DNA binding motif first found in Doublesex of Drosophila and mab-3 of Caenorhabditis elegans, regulate sexual differentiation in multiple phyla. The DM domain gene Dmrt1 is essential for testicular differentiation in vertebrates. In the mouse, Dmrt1 is expressed in pre-meiotic germ cells and in Sertoli cells, which provide essential support for spermatogenesis. Dmrt1 null mutant mice have severely dysgenic testes in which Sertoli cells and germ cells both fail to differentiate properly after birth. Here we use conditional gene targeting to identify the functions of Dmrt1 in each cell type. We find that Dmrt1 is required in Sertoli cells for their postnatal differentiation, and for germ line maintenance and for meiotic progression. Dmrt1 is required in germ cells for their radial migration to the periphery of the seminiferous tubule where the spermatogenic niche will form, for mitotic reactivation and for survival beyond the first postnatal week. Thus Dmrt1 activity is required autonomously in the Sertoli and germ cell lineages, and Dmrt1 activity in Sertoli cells is also required non-autonomously to maintain the germ line. These results demonstrate that Dmrt1 plays multiple roles in controlling the remodeling and differentiation of the juvenile testis.     

In vitro generation of germ cells from murine embryonic stem cells

The demonstration of germ cell and haploid gamete development from embryonic stem cells (ESCs) in vitro has engendered a unique set of possibilities for the study of germ cell development and the associated epigenetic phenomenon. The process of embryoid body (EB) differentiation, like teratoma formation, signifies a spontaneous differentiation of ESCs into cells of all three germ layers, and it is from these differentiating aggregates of cells that putative primordial germ cells (PGCs) and more mature gametes can be identified and isolated. The differentiation system presented here requires the differentiation of murine ESCs into EBs and the subsequent isolation of PGCs as well as haploid male gametes from EBs at various stages of differentiation. It serves as a platform for studying the poorly understood process of germ cell allocation, imprint erasure and gamete formation, with 4-6 weeks being required to isolate PGCs as well as haploid cells.     

PRDM在生殖细胞发育中的作用

生殖细胞特化是发育和遗传的基础。原始生殖细胞（精子和卵子的前体细胞）的特化包括3个主要事件：体细胞程序的抑制、潜在全能性的获得、基因组范围内的表观遗传重编程。含PR域蛋白1（PR domain-containing1,PRDM1）和PRDM14是生殖细胞系产生的关键转录调节因子。PRDMl要抑制体细胞程序,而PRDM14主要调节潜在全能性的获得及表观遗传学重编程。此外,PRDM家族蛋白PRDM9在生殖细胞减数分裂中有重要作用。     

A selective screen reveals discrete functional domains in Drosophila Nanos

The Drosophila protein Nanos encodes an evolutionarily conserved protein with two zinc finger motifs. In the embryo, Nanos protein function is required for establishment of the anterior-posterior body pattern and for the migration of primordial germ cells. During oogenesis, Nanos protein is involved in the establishment and maintenance of germ-line stem cells and the differentiation of oocyte precursor cells. To establish proper embryonic patterning, Nanos acts as a translational regulator of hunchback RNA. Nanos' targets for germ cell migration and development are not known. Here, we describe a selective genetic screen aimed at isolating new nanos alleles. The molecular and genetic analysis of 68 new alleles has allowed us to identify amino acids critical for nanos function. This analysis shows that the CCHC motifs, which coordinate two metal ions, are essential for all known functions of Nanos protein. Furthermore, a region C-terminal to the zinc fingers seems to constitute a novel functional domain within the Nanos protein. This "tail region" of Nanos is required for abdomen formation and germ cell migration, but not for oogenesis.     

Function of cyclins in regulating the mitotic and meiotic cell cycles in male germ cells

The unique cell cycles that characterize various aspects of the differentiation of germ cells provide a unique opportunity to understand heretofore elusive aspects of the in vivo function of cell cycle regulators. Key components of the cell cycle machinery are the regulatory sub-units, the cyclins, and their catalytic partners, the cyclin-dependent kinases. Some of the cyclins exhibit unique patterns of expression of germ cells that suggest possible concomitant distinct functions, predictions that are being explored by targeted mutagenesis in mouse models. A novel, meiosis-specific function has been shown for one of the A-type cyclins, cyclin A1. Embryonic lethality has obviated understanding of the germline functions of cyclin A2 and cyclin B1, while yet other cyclins, although expressed at specific stages of germ cell development, may have less essential function in the male germline.     

胚胎干细胞诱导分化为雄性生殖细胞的研究进展

胚胎干细胞（embryonic stem cells,ES细胞）具有自我更新及无限分化潜能,理论上可以分化为生殖细胞。目前,在人及鼠中已有体外诱导ES细胞分化为成熟精子的报道。系统阐述影响ES细胞分化为雄性生殖细胞的内源性及外源性因素,并结合国内外最新研究进展总结其诱导分化方法,展望应用前景,期望为从事相关研究的学者提供参考。     

NOTCH1 Gain of Function in Germ Cells Causes Failure of Spermatogenesis in Male Mice

NOTCH1 is a member of the NOTCH receptor family, a group of single-pass trans-membrane receptors. NOTCH signaling is highly conserved in evolution and mediates communication between adjacent cells. NOTCH receptors have been implicated in cell fate determination, as well as maintenance and differentiation of stem cells. In the mammalian testis expression of NOTCH1 in somatic and germ cells has been demonstrated, however its role in spermatogenesis was not clear. To study the significance of NOTCH1 in germ cells, we applied a cre/loxP approach in mice to induce NOTCH1 gain- or loss-of function specifically in male germ cells. Using a Stra8-icre transgene we produced mice with conditional activation of the NOTCH1 intracellular domain (NICD) in germ cells. Spermatogenesis in these mutants was progressively affected with age, resulting in decreased testis weight and sperm count. Analysis of downstream target genes of NOTCH1 signaling showed an increased expression of Hes5, with a reduction of the spermatogonial differentiation marker, Neurog3 expression in the mutant testis. Apoptosis was significantly increased in mouse germ cells with the corresponding elevation of pro-apoptotic Trp53 and Trp63 genes'' expression. We also showed that the conditional germ cell-specific ablation of Notch1 had no effect on spermatogenesis or male fertility. Our data suggest the importance of NOTCH signaling regulation in male germ cells for their survival and differentiation.