CONTROLLING MECHANISMS IN MAMMALIAN REPRODUCTION

该计划项目的总体目标是阐明复杂的 细胞和遗传控制机制之间的相互作用 它们调节哺乳动物发育的早期阶段。在.期间 从受精卵到卵子的一系列卵裂分裂 胚泡，胚胎基因组的顺序激活发生 伴随着第一个明显的分化迹象 滋养外胚层和胚胎组织的起源。由 囊胚期，胚胎包含所有的谱系前体 胚胎和胚外组织。确定…的性质 这些早期和关键的差异化事件，我们建议 以下是实验方法： 1)通过所有调查人员的共同努力，基因探针 它们反映了每个发育阶段的基因表达将 被开发出来。这些探头将用于识别 相应的基因产物并开始分析它们的 功能。 2)通过瞬时或永久地将特定的 基因和/或其产物，这些基因的功能作用 将会被确定。 3)将对核移植胚胎进行分析以确定 他们发育失败的原因以及确定 基因参与其中。此方法的一部分将涉及分析 配子发生过程中的特异性和基因调控。 4)生长控制分子(包括生长)的作用 调节生长的因子及其受体和癌基因) 早期胚胎的分化模式包括 将对滋养外胚层和内细胞团进行研究。的作用 蛋白质磷酸化在转录调控中的作用 将检测胚胎基因的活性。 5)细胞谱系的建立和细胞谱系特异性 通过注射合适的DNA来探索基因的表达 标记和与特定探针的原位杂交。 6)滋养外胚层、滋养层细胞和内细胞的分化 将通过鉴定基因和基因产物来研究质量 通过确定它们的作用和通过 企图首先明确地改变或废除这一点 小鼠胚胎发生中的分化事件。 鉴于滋养外胚层组织的可及性，很容易 涉及的发育事件的特征序列 滋养外胚层形成。

The overall goal of the Program Project is to elucidate the complex interactions between the cellular and genetic control mechanisms which regulate the early stages of mammalian development. During the series of cleavage division which leads from zygote to blastocyst, sequential activation of the embryonic genome occurs concomitantly with the first overt signs of differentiation of trophectoderm and the origins of embryonic tissues. By the blastocyst stage, the embryo contains the lineage precursors of all embryonic and extraembryonic tissues. To determine the nature of these early and crucial differentiation events, we propose the following experimental approaches: 1) Through the joint effort of all investigators, genetic probes which reflect the gene expression at each developmental stage will be developed. These probes will be used to identify the corresponding gene products and to begin the analysis of their function. 2) By transient or permanent introduction into embryos of specific genes and/or their products, the functional role of these genes will be determined. 3) Nuclear transfer embryos will be analysed to determine the reasons for their developmental failure and for identifying the genes involved. Part of this approach will involve analysis of specific and gene regulation during gametogenesis. 4) The role of growth controlling molecules (including growth factors and their receptors and oncogenes) in regulating the growth and differentiation patterns in early embryos including trophectoderm and inner cell mass will be studied. The role of protein phosphorylation in regulating the transcriptional activity of the embryonic gene will be examined. 5) Establishment of cell lineages and the cell lineage specific gene expression will be explored by injection of suitable DNA markers and by in situ hybridization with specific probes. 6) Differentiation of trophectoderm, trophoblast and inner cell mass will be studied by identifying genes and gene products specific for those tissues, by determining their role and by attempting to specifically alter or abolish this first differentiation event in mouse embryogenesis. In view of the accessibility of trophectoderm tissue, the readily characterized sequence of developmental events involved in trophectoderm formation.