ISOLATION OF THE NEURONAL GENE WEAVER

神经元基因编织者的分离

• 批准号: 2269626
• 负责人: SUZANNE Katherine ROFFLER-TARLOV
• 金额: $ 25.69万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-08 至 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2269626
• 关键词: Parkinson's disease; animal genetic material tag; artificial chromosomes; autosomal recessive trait; cerebellum; complementary DNA; disease /disorder model; gene expression; gene mutation; genetic markers; granule cell; hybrid cells; laboratory mouse; linkage mapping; mesencephalon; neural degeneration; neurogenesis; nucleic acid repetitive sequence; protein structure function; restriction fragment length polymorphism; sequence tagged sites; southern blotting; structural genes; subtraction hybridization; transfection

This revised proposal requests support for the cloning and characterization of the murine mutant gene weaver (wv) and its normal allele. Weaver is an autosomal recessive gene that very selectively disrupts neuronal differentiation and causes cell death in the cerebellum and in the midbrain. Evidence to date points to wv causing deficits in neurite formation in both the dopamine- containing neurons of the midbrain and the granule cells of the cerebellum. These deficits lead to arrested neuronal migration of cerebellar granule cells and their eventual death and to death of a subset of dopamine-containing neurons in the substantia nigra, pars compacta; the mouse homologues of the neurons that die in Parkinson's disease patients. A unique feature of this plan is its pairing of advantages of functional cloning with locational cloning methods. The strategy is designed to identify candidates for wv and its normal allele by isolation of genomic and coding sequences from l) the chromosomal wv region (chromosome 2l in human and l6 in mouse) by YAC technology and 2) brain sources enriched for wv transcripts by a variety of subtractive libraries. The candidates derived from each method will be quickly screened for appropriate chromosomal location using somatic cell hybrids to identify candidates that map to distal chromosome l6. Those passing this screen will be subsequently screened on Southern blots from our linkage assay to identify candidates that map to the wv region. Only these candidate gene(s) will be tested for ability to rescue the defective wv phenotype in an in vitro functional assay. The genes discovered as a result of this research are likely to be part of a cascade necessary for neuronal differentiation as well as part of the system that is altered in Parkinson's disease and hereditary ataxias that involve the dopamine-containing pathways. Certainly, the identification of genes and proteins that maintain the neurons that die in these diseases would open new possibilities for treatment.

此修订的建议要求对克隆的支持和 鼠突变基因编织器（WV）及其的表征 正常的等位基因。 织布工是一个非常非常的常染色体隐性基因 有选择地破坏神经元分化并导致细胞死亡 在小脑和中脑。 到目前为止的证据指向 WV在多巴胺中引起神经突的缺陷 - 包含中脑的神经元和颗粒细胞的神经元 小脑。 这些赤字导致被捕的神经元迁移 小脑颗粒细胞及其最终死亡以及死亡 黑质中含有多巴胺神经元的一部分， pars commacta;死亡神经元的老鼠同源物 帕金森氏病患者。 该计划的一个独特功能是其优势配对 使用位置克隆方法的功能克隆。 策略 旨在通过通过 从L）染色体分离基因组和编码序列 YAC的WV区域（人类中的染色体2L和L6中的L6染色体） 技术和2）大脑来源丰富了WV成绩单 各种减法库。 候选人源自每个 方法将快速筛选以获取适当的染色体 使用体细胞杂种来识别映射的候选者的位置 远端染色体L6。 那些通过此屏幕的人将是 随后从我们的链接测定到南部印迹筛选 确定映射到WV区域的候选人。 只有这些 候选基因将进行测试，以挽救能力 体外功能测定中的WV表型有缺陷。 这项研究结果发现的基因可能是 神经元分化所必需的级联的一部分 帕金森氏病和 涉及含多巴胺的途径的遗传性障碍。 当然，维持基因和蛋白质的鉴定 在这些疾病中死亡的神经元将开放新的可能性 用于治疗。