GENE EXPRESSION IN EARLY EMBRYONIC DEVELOPMENT

早期胚胎发育中的基因表达

• 批准号: 2403086
• 负责人: FRED H WILT
• 金额: $ 28.79万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-09-01 至 1999-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2403086
• 关键词: DNA binding protein; alternatives to animals in research; cell cell interaction; cell differentiation; developmental genetics; early embryonic stage; gene expression; genetic library; genetic markers; genetic promoter element; genetic transcription; immunocytochemistry; in situ hybridization; molecular cloning; nonmammalian vertebrate embryology; nucleic acid sequence; oogenesis; protein biosynthesis; radiotracer; saltwater environment; sea urchins; tissue /cell preparation

The broad objective of this proposal is to learn how cells interact in the developing embryo to bring about determination and differentiation. the commitment of the progeny of a totipotent ovum to the various differentiation pathways of the developing embryo is a basic biological problem of prime importance. Choices made by cells to follow different paths are central to normal and abnormal embryonic development, to cancer, would repair, and aging. The experiments will focus on the sea urchin embryo, a system very similar to vertebrates in its reliance on cell interactions as a way of bringing about determination of various cell types and lineages in early development. This embryo has various blastomeres that can be separated form one another and recombined in various ways. New tissue specific markers that serve as reliable indicators of specific differentiation pathways are now available. first, these markers will be used to learn the rules that govern specific gene expression. This will involve experiments in which individual blastomeres are cultured alone or in combination with other blastomere types, or with various agents (e.g., Li ion, FGF). The fate of the blastomeres and expression of various genes will be measured by in situ hybridization and immunocytochemistry. Are there localized substances in the vegetal hemisphere of the egg that initiate the lineages that form gut and spicules: Are there ions or agonists that might be effective in causing vegetal differentiation of animal cells? Do these agents use secondary messenger pathways to effect their actions? Second, the effort to develop and characterize tissue specific cDNA clones will be continued. The efforts will concentrate on cloning spicule matrix genes, and on genes expressed solely in descendants of mesomeres. Third, the details of how expression of spicule matrix genes is regulated will be studied. Transcription of these genes will be studied, in vitro, in nuclear extracts that transcribe defined templates. This functional assay will be used to conduct studies on the DNA sequences necessary for promoter activity and to study proteins in the extracts that interact with the DNA.

该提案的主要目标是了解细胞如何相互作用 发育中的胚胎带来决定和分化。 全能卵子的后代对各种 发育中胚胎的分化途径是基本的生物学途径 的首要问题。 细胞做出的选择遵循不同的 路径对于正常和异常胚胎发育至关重要 癌症、修复和衰老。 实验将集中在海洋 海胆胚胎，一个与脊椎动物非常相似的系统，依赖于 细胞相互作用作为决定各种因素的一种方式 早期发育中的细胞类型和谱系。 这个胚胎有多种 卵裂球可以彼此分离并重新组合 各种方式。 新的组织特异性标记物可作为可靠的 现在可以获得特定分化途径的指标。 首先，这些标记将用于学习管理规则 特定的基因表达。 这将涉及实验，其中 单个卵裂球单独培养或与其他卵裂球联合培养 卵裂球类型，或使用各种试剂（例如锂离子、FGF）。 命运 卵裂球的数量和各种基因的表达将通过以下方式测量 原位杂交和免疫细胞化学。 有本地化的吗 卵子植物半球中启动卵子发育的物质 形成肠道和针状体的谱系：是否有离子或激动剂 可能有效引起动物细胞的植物分化？ 这些药物是否使用第二信使途径来影响它们的作用？ 行动？ 其次，努力开发和表征组织特异性 cDNA克隆将继续进行。 努力将集中在克隆上 针状基质基因，以及仅在后代中表达的基因 中粒。 三、针状基质基因如何表达的细节 受到监管，将进行研究。 这些基因的转录将是 在体外研究了转录定义的核提取物 模板。 该功能测定将用于进行研究 启动子活性和研究蛋白质所必需的 DNA 序列 与 DNA 相互作用的提取物。