MESSENGER RNA DISTRIBUTION IN SEA URCHIN EMBRYOS

海胆胚胎中信使 RNA 的分布

• 批准号: 3073176
• 负责人: ROBERT C ANGERER
• 金额: $ 5.41万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-08-01 至 1989-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3073176
• 关键词: actins; animal population genetics; cell differentiation; developmental genetics; early embryonic stage; gene expression; genetic manipulation; genetic mapping; genetic recombination; histones; in situ hybridization; messenger RNA; nonmammalian vertebrate embryology; nucleic acid hybridization; nucleic acid sequence; radiotracer; saltwater environment; sea urchins; tubulin

The aim of the proposed research is to determine the specificity of expression of individual genes in various cell lineages of the sea urchin (Stronglylocentrotus purpuratus) embryo. In situ hybridization techniques will be used to detemine the spatial distribution of individual mRNAs, using radioactively labeled RNA probes derived from recombinant DNAs containing individual sea urchin mRNA sequences. Using probes representing mRNAs expressed at blastula stage, the extent of restriction of individual mRNAs to one or more cell lineages during development will be examined, and the time of appearance of specific mRNA sets in different lineages will be analyzed. Cell-lineage-specific mRNAs will be used as markers to construct a fate map of the embryo at the level of mRNA expression. The tissue specificity of expression of different members of 3 multi-gene families will be investigated. Detaled patterns will be established for 6 actin genes which preliminary studies have shown are expressed in a highly lineage- and stage-specific manner. The timing of the switch in expression of histone genes from early to late variants will be compared in different cell lineages. Possible correlations between this switch and other developmental events (expression of cell-type-specific mRNAs, changes in rate of cell division) will be examined. The distributions of different late-variant histone mRNAs of the H2B subclass will be compared. Spatial patterns of expression of Alpha and Beta tubulin mRNAs will be investigated. The timing of appearance of cell-lineage-specific mRNAs will be related to events of differentiation. The correlation of different pathways of differentiation with expression of specific mRNA sets will be examined using procedures which alter the determination/differentiation of cell lineages. These studies are fundamental to understanding the requirements for differential gene expression during early development.

拟议研究的目的是确定 海胆不同细胞谱系中单个基因的表达 （Stronglylocentrotus purpuratus）胚胎。 原位杂交技术 将用于确定单个 mRNA 的空间分布， 使用源自重组 DNA 的放射性标记 RNA 探针 含有单个海胆 mRNA 序列。 使用探针代表 囊胚期表达的mRNA，个体的限制程度 将检查发育过程中一种或多种细胞谱系的 mRNA，并且 不同谱系中特定 mRNA 组出现的时间将是 分析了。 细胞谱系特异性 mRNA 将用作标记来构建 mRNA 表达水平上的胚胎命运图。 组织 3个多基因家族不同成员的表达特异性 将被调查。 将为 6 个肌动蛋白建立详细的模式 初步研究表明基因高度表达 谱系和阶段特定的方式。 表达切换的时机 组蛋白基因从早期到晚期的变异将在不同的情况下进行比较 细胞谱系。 此开关与其他开关之间可能存在的关联 发育事件（细胞类型特异性 mRNA 的表达、 细胞分裂率）将被检查。 不同的分布 将比较 H2B 亚类的晚期变异组蛋白 mRNA。 空间 Alpha 和 Beta 微管蛋白 mRNA 的表达模式将是 调查了。 细胞谱系特异性 mRNA 出现的时间将 与分化事件有关。 不同的相关性 具有特定 mRNA 组表达的分化途径将是 使用改变测定/区分的程序进行检查 细胞谱系。 这些研究对于理解 早期发育过程中差异基因表达的要求。