DEVELOPMENTAL GENETICS OF NEURAL TUBE FORMATION

神经管形成的发育遗传学

• 批准号: 3317919
• 负责人: RALPH J GREENSPAN
• 金额: $ 16.09万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-04-01 至 1987-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3317919
• 关键词: beta galactosidase; congenital nervous system disorder; developmental genetics; early embryonic stage; embryo /fetus; gene interaction; gene mutation; genetic mapping; genetic markers; homozygote; linkage mapping; messenger RNA; mutant

This study will probe the role of gene interactions in mouse neural tube development. Formation of the neural tube is one of the first, most critical events in mammalian embryogenesis, and defects in the process (including genetic ones) are among the most common human birth defects. The study will make use of three newly induced mutations in the mouse, isolated on the basis of their interactions with loci known to affect neural tube development. These new mutants will be characterized genetically and developmentally, determining their morphological effects in embryos and the modifications they impose on other neural tube mutations already described. Studies will include linkage and mapping, histological analysis of embryos, construction of double mutants and determination of lethal phases. The new mutants were isolated for their interactions with the Splotch locus, which causes failure of neural tube closure in homozygous condition, and Fused, which produces duplications of the neural tube in homozygotes. The study of new interacting loci will help define the set of genes whose coordinate action is required for normal neural tube formation. Since many of the genes that direct neural tube development are likely to mediate cell interactions, it will be essential to establish if a mutant defect is intrinsic to cells of the neural plate, or whether it results from an abnormal cell interaction of extrinsic origin. In order to address this question, we will construct genetic mosaics by aggregating mutant and wild-type embryos. The cells' genotypes will be scored with a new molecular marker, a gene fusion placing bacterial Beta-galactosidase under the control of a eukaryotic promoter, which will be introduced into the germ line of a normal mouse strain. Thus, it will be possible to score the distribution of mutant vs. wild-type cells, using antibody staining of sectioned embryos. In this manner, a correlation can be made as to which cells must carry a particular mutation in order to produce the mutant phenotype, defining the cellular site of action of a given mutation. This technique will be used to map the various genetic and tissue contributions to neural tube development, as well as to trace the cell lineage of the neural tube. In the long run, studies of this sort will define the set of genes required for neural tube formation and lay the foundations for molecular studies of this process.

这项研究将探讨基因相互作用在小鼠神经管中的作用 发展。 神经管的形成是最早也是最重要的过程之一 哺乳动物胚胎发生中的关键事件以及过程中的缺陷 （包括遗传缺陷）是最常见的人类出生缺陷之一。 该研究将利用小鼠体内三种新诱导的突变， 根据它们与已知影响的位点的相互作用进行分离 神经管发育。 这些新突变体将被表征 遗传和发育，确定它们的形态效应 胚胎及其对其他神经管突变施加的修饰 已经描述过。 研究将包括连接和绘图、组织学 胚胎分析、双突变体构建和测定 致死阶段。 新突变体因其与 Splotch 基因座，导致神经管闭合失败 纯合条件和融合，产生神经网络的重复 纯合子中的管。 新的相互作用基因座的研究将有助于定义 正常神经管需要其协调作用的一组基因 形成。 由于许多指导神经管发育的基因可能 介导细胞相互作用，因此必须确定突变体是否 缺陷是神经板细胞固有的，还是它的结果 来自外源性异常细胞相互作用。 为了解决 这个问题，我们将通过聚合突变体和 野生型胚胎。 细胞的基因型将用新的评分 分子标记，将细菌 β-半乳糖苷酶置于 真核启动子的控制，将被引入 正常小鼠品系的种系。 如此一来，就有可能获得 突变型与野生型细胞的分布，使用抗体染色 切片胚胎。 以这种方式，可以建立以下相关性： 细胞必须携带特定的突变才能产生突变体 表型，定义给定突变的细胞作用位点。 这 技术将用于绘制各种遗传和组织贡献图 神经管发育，以及追踪细胞谱系 神经管。 从长远来看，此类研究将确定所需的基因组 神经管形成并为分子研究奠定基础 这个过程。