原始生殖细胞迁移的研究进展

概述了不同类群动物原始生殖细胞（primordial germ cells,PGCs）的起源和迁移路径.介绍了当前有关不同类群动物PGCs在胚胎发育中与迁移有关的因子和基因的功能,这些因子和基因包括细胞外基质中的生物大分子纤粘连蛋白（fibrbonectin）、Tenascin-C、层粘连蛋白（laminin）,Foxc1基因、JAK/STAT信号通路、信号分子STAT 和RAS、基质细胞衍生因子-1（stromal cell-derived factor-1,SDF-1）、F-肌动蛋白（F-actin）等.评述了PGCs迁移研究中存在的问题,认为众多调控因子与PGCs迁移的关系虽已被揭示,但各因子对PGCs详细作用机制及其相互联系、不同物种PGCs迁移的调控因子之间的同源性及如何把单一的作用效果整合起来等问题有待解决.     

Long-term proliferation of mouse primordial germ cells in culture.

Primordial germ cells (PGCs) are first identifiable as a population of about eight alkaline phosphatase-positive cells in the 7.0 days postcoitum mouse embryo. During the next 6 days of development they proliferate to give rise to the 25,000 cells that will establish the meiotic population. Steel factor is required for PGC survival both in vivo and in vitro and together with leukaemia inhibitory factor stimulates PGC proliferation in vitro. In feeder-dependent culture, PGCs will proliferate for up to 7 days, but their numbers eventually decline and their proliferative capacity is only a fraction of that seen in vivo. Here we report a further factor that stimulates PGC proliferation in vitro, basic fibroblast growth factor (bFGF). Furthermore, bFGF, in the presence of steel factor and leukaemia inhibitory factor, stimulates long-term proliferation of PGCs, leading to the derivation of large colonies of cells. These embryonic germ cells resemble embryonic stem cells, pluripotent cells derived from preimplantation embryos, or feeder-dependent embryonal carcinoma cells, pluripotent stem cells of PGC-derived tumours (teratomas and teratocarcinomas). To our knowledge, these results provide the first system for long-term culture of PGCs.     

Effects of the steel gene product on mouse primordial germ cells in culture.

Mutations at the steel (sl) and dominant white spotting (W) loci in the mouse affect primordial germ cells (PGC), melanoblasts and haemopoietic stem cells. The W gene encodes a cell-surface receptor of the tyrosine kinase family, the proto-oncogene c-kit. In situ analysis has shown c-kit messenger RNA expression in PGC in the early genital ridges. The Sl gene encodes the ligand for this receptor, a peptide growth factor, called here stem cell factor (SCF). SCF mRNA is expressed in many regions of the early mouse embryo, including the areas of migration of these cell types. It is important now to identify the role of the Sl-W interaction in the development of these migratory embryonic stem cell populations. Using an in vitro assay system, we show that SCF increases both the overall numbers and colony sizes of migratory PGC isolated from wild-type mouse embryos, and cultured on irradiated feeder layers of STO cells (a mouse embryonic fibroblast line). In the absence of feeder cells, SCF causes a large increase in the initial survival and apparent motility of PGC in culture. But labelling with bromodeoxyuridine shows that SCF is not, by itself, a mitogen for PGC. SCF does not exert a chemotropic effect on PGC in in vitro assays. These results suggest that SCF in vivo is an essential requirement for PGC survival. This demonstrates the control of the early germ-line population by a specific trophic factor.     

Effect of Steel factor and leukaemia inhibitory factor on murine primordial germ cells in culture.

Despite the importance of germ cells to the survival of species, surprisingly little is known about their embryological origin, proliferation, migration and entry into mitotic arrest or meiosis. Mutations in the murine Dominant White Spotting (W) and Steel genes, which respectively encode the c-kit tyrosine kinase receptor and the c-kit ligand (or Steel factor), impair the development of primordial germ cells (PGCs) in vivo, as well as haematopoietic stem cells and neural crest-derived melanoblasts. Here we use a monoclonal antibody against c-kit tyrosine kinase receptor and recombinant Steel factor to study the c-kit receptor-ligand system in cultured PGCs. In addition, we show that leukaemia inhibitory factor (also known as differentiation inhibitory activity), a factor secreted by STO fibroblasts, can stimulate proliferation of primordial germ cells in vitro.     

转基因食品安全的生态伦理学探析

转基因食品已经存在了十几年,但人们对它安全的争议仍然颇多.与此同时,转基因作物种植面积又在逐年扩大.转基因食品应该如何发展?笔者从生态伦理学角度,以生态化生存模式为背景,在重新认识转基因食品安全及发展问题的基础上,探讨相应对策.     

转基因鱼生态效应的评价

转基因鱼的优良经济性状为水产业提供了巨大的发展潜力，但转基因鱼释放到自然环境后的生态效应则引起了很多争议。文章分析了转基因鱼的生态风险，并讨论了特洛伊基因效应模型。考虑到环境中各个生态因子对生物体表型的影响，生态学家在对转基因鱼进行生态风险的评价时，不能仅仅依赖于实验室的研究和理论模型，而需要建立人工模拟生态系统，从而得到全面、可靠的结论。     

消费者对转基因食品标识认知状况分析

在广东省佛山市进行的一项关于消费者对转基因食品消费倾向的调查中发现,有16.5%的受访者不了解转基因食品,33.6%的受访者不知道我国已经对转基因产品实行标识制度,42.2%的受访者虽然知道有标识制度但不能辨认转基因产品标识。调查结果表明,市民对转基因食品的认知,尤其是对转基因食品标识制度的认知仍然缺乏。因此,必须进一步加强和改进对转基因生物的管理,加大对转基因食品相关知识的宣传,以便保障消费者的知情权和选择权。     

Requirement for mast cell growth factor for primordial germ cell survival in culture.

Mast-cell growth factor (MGF) is encoded by the murine steel (Sl) locus and is a ligand for the tyrosine kinase receptor protein encoded by the proto-oncogene c-kit at the murine dominant white spotting (W) locus. Mutations at both these loci affect mast cells, primordial germ cells (PGCs), haemopoietic stem cells and melanocytes. In many Sl and W mutants, the rapid proliferation of PGC that normally occurs between day 7 and 13.5 of embryonic development fails to occur. As c-kit is expressed in PGCs while MGF is expressed in the surrounding mesenchyme, MGF might promote the proliferation of PGCs. Here we report that MGF is essential for PGC survival in culture, but does not stimulate PGC proliferation. Moreover, whereas both the transmembrane and soluble proteolytic cleavage forms of MGF stimulate mast-cell proliferation, soluble MGF has a relatively limited ability to support survival of PGCs in culture, thus explaining the sterility in mice carrying the steel-dickie (Sld) mutation, which encodes only a soluble form of MGF, and providing a functional role for a transmembrane growth factor.     

Gene flow from genetically modified herbicide-resistant rapeseed to cruciferous weeds

The sexual compatibility between genetically modified (GM) glyphosate-resistant rapeseed variety Q3 (Brassica napus L.) and 5 cruciferous weeds is studied through the observation of fluorescence microscopy and cross-fertility after manual pollination. The results indicated that Q3 (as male) was highly incompatible with Thlaspi arvense L., Capsella bursa?pastoris (L.) Medic, Cardamine hirsuta L. and Rorippa palustris (L.) Besser (as female). Fluorescence microscopic observation showed that growing of pollen tubes terminated on the stigma surface or at the upper 1/3 part of the style. However, B. juncea×Q3 was compatible, and the compatibility index was 1.65. Under the neighboring growth and natural pollination conditions, the rates of gene flow from Q3 to T. arvense, C. bursa-pastoris, C. hirsute and R. palustris were all 0, while it was 0.86% for B. juncea. These results indicate that there is difference in the rate of gene flow between GM rapeseed and cruciferous wild weeds, and frequency of gene flow is highly correlated with sexual compatibility.     

Detection of genetically modified ingredients in seed samples from Heilongjiang soybean field

A total of 210 soybean samples collected from different areas of Heilongjiang Province were analyzed by PCR to detect the presence of RR (Roundup Ready) soybean ingredient. Results showed that CaMV35S promoter was not detected in all samples. CP4-EPSPS gene was amplified in 13 samples, but all of them were proved to be false positives in restriction endonuclease digestion analysis of PCR products. Further analysis by nested PCR indicated that there were no RR soybean ingredients in the samples. Besides, the amplified fragments of 0xl by CP4-EPSPS primers were sequenced, and the results confirmed again the fact that this fragment was not from CP4-EPSPS gene.     

Germline stem cells and follicular renewal in the postnatal mammalian ovary

A basic doctrine of reproductive biology is that most mammalian females lose the capacity for germ-cell renewal during fetal life, such that a fixed reserve of germ cells (oocytes) enclosed within follicles is endowed at birth. Here we show that juvenile and adult mouse ovaries possess mitotically active germ cells that, based on rates of oocyte degeneration (atresia) and clearance, are needed to continuously replenish the follicle pool. Consistent with this, treatment of prepubertal female mice with the mitotic germ-cell toxicant busulphan eliminates the primordial follicle reserve by early adulthood without inducing atresia. Furthermore, we demonstrate cells expressing the meiotic entry marker synaptonemal complex protein 3 in juvenile and adult mouse ovaries. Wild-type ovaries grafted into transgenic female mice with ubiquitous expression of green fluorescent protein (GFP) become infiltrated with GFP-positive germ cells that form follicles. Collectively, these data establish the existence of proliferative germ cells that sustain oocyte and follicle production in the postnatal mammalian ovary.     

Chromatin remodelling and epigenetic features of germ cells

Germ cells have the unique capacity to start a new life upon fertilization. They are generated during a sex-specific differentiation programme called gametogenesis. Maturation of germ cells is characterized by an impressive degree of cellular restructuring and gene regulation that involves remarkable genomic reorganization. These events are finely tuned, but are also susceptible to the introduction of various types of error. Because stable genetic transmission to future generations is essential for life, understanding the control of these processes has far-reaching implications for human health and reproduction.     

Derivation of embryonic germ cells and male gametes from embryonic stem cells

Egg and sperm cells (gametes) of the mouse are derived from a founder population of primordial germ cells that are set aside early in embryogenesis. Primordial germ cells arise from the proximal epiblast, a region of the early mouse embryo that also contributes to the first blood lineages of the embryonic yolk sac. Embryonic stem cells differentiate in vitro into cystic structures called embryoid bodies consisting of tissue lineages typical of the early mouse embryo. Because embryoid bodies sustain blood development, we reasoned that they might also support primordial germ cell formation. Here we isolate primordial germ cells from embryoid bodies, and derive continuously growing lines of embryonic germ cells. Embryonic germ cells show erasure of the methylation markers (imprints) of the Igf2r and H19 genes, a property characteristic of the germ lineage. We show that embryoid bodies support maturation of the primordial germ cells into haploid male gametes, which when injected into oocytes restore the somatic diploid chromosome complement and develop into blastocysts. Our ability to derive germ cells from embryonic stem cells provides an accessible in vitro model system for studies of germline epigenetic modification and mammalian gametogenesis.