GENE EXPRESSION DURING MURINE NEURAL TUBE CLOSURE

小鼠神经管闭合期间的基因表达

• 批准号: 2156350
• 负责人: RICHARD H. FINNELL
• 金额: $ 17.23万
• 依托单位: TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-07-01 至 1997-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2156350
• 关键词: antisense nucleic acid; congenital disorders; embryo /fetus cell /tissue; fusion failure; gene expression; genetic transcription; growth factor; growth factor receptors; homeobox genes; in situ hybridization; laboratory mouse; messenger RNA; molecular pathology; natural gene amplification; neural plate /tube; neurogenesis; northern blottings; nucleic acid hybridization; nucleic acid probes; statistics /biometry

Neural tube defects are amongst the most common of all birth defects in humans, affecting approximately 1 in 1,000 live born infants. These malformations, which include spina bifida, anencephaly and meningomyeloceles, represents the single largest group of etiologies for patients with mental deficiency. In spite of the common nature of these congenital defects, efforts to obtain a clear understanding of their pathogenesis on a molecular level have been limited by the availability of sufficient quantities of embryonic material from which to isolate mRNA and ultimately develop cDNA libraries to screen. The research program described in this proposal takes advantage of two recent advances in molecular biological techniques that can overcome this problem. Using in situ transcription (IST) and anti-sense RNA (aRNA) amplification, we intend to examine in murine embryos the composite changes in a population of mRNAs from selected candidate genes whose function is believed to be critical for normal neural tube morphogenesis. The experimental paradigm will focus on the curly tail (ct) mutant, but it will also include embryos bearing a genetic mutation at the Splotch locus (Sp), which confers spontaneous anterior and/or posterior neural tube defects in homozygous embryos. These experiments should provide new data on gene expression during the period of neural tube closure in embryos with neural tube defects. Utilizing a differential hybridization screening strategy, it will be possible to isolate those sequences that are vastly enriched or in very low abundance yet remain of critical important in the pathogenesis of neural tube defects. It will also be possible to determine whether anterior and posterior defects are mechanistically unique, or if they merely represent regional differences of the same malformation. The results obtained in these murine models will provide insight into the development of analogous defects in humans, which may lead to better preventative measures.

神经管缺陷是所有先天缺陷中最常见的缺陷之一 人类影响了大约1,000名活着的婴儿中的大约1个。这些 畸形，包括脊柱裂，阿诺班和 脑膜瘤球代表了最大的病因群 精神缺乏的患者。尽管有这些共同的本质 先天性缺陷，为清楚了解他们的努力 分子水平上的发病机理已受到限制 足够数量的胚胎材料，从中分离出mRNA和 最终开发cDNA库进行筛选。研究计划 该提案中描述的利用了两个最新进展 可以克服这个问题的分子生物学技术。使用 原位转录（IST）和抗沉思RNA（ARNA）扩增，我们 打算在鼠类胚胎中检查人群的综合变化 来自选定候选基因的mRNA的功能被认为是 对于正常的神经管形态发生至关重要。实验范式 将专注于卷曲尾部（CT）突变体，但也将包括胚胎 在斑点基因座（SP）上带有遗传突变 纯合的自发前和/或后神经管缺陷 胚胎。这些实验应提供有关基因表达的新数据 在神经管中的神经管闭合期间 缺陷。利用差异杂交筛选策略， 可以隔离那些非常丰富或在内的序列 非常低的丰度，但在发病机理中仍然至关重要 神经管缺陷。还可以确定是否 前后缺陷在机械上是独一无二的，或者如果它们 仅代表相同畸形的区域差异。这 在这些鼠模型中获得的结果将提供有关 人类类似缺陷的发展，这可能会导致更好 预防措施。