NEUROBIOLOGY OF DEVELOPMENT

发育的神经生物学

• 批准号: 3100080
• 负责人: WILLIAM T NORTON
• 金额: $ 77.29万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-04-01 至 1995-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3100080
• 关键词: developmental neurobiology

This program contains four projects devoted to the neurochemistry and cell biology of nervous system development. All studies are to be performed in rats. We propose: (1) to isolate and purify glial precursor cells from the developing and mature rat brain, to follow the differentiation of these cells in culture, and to test the hypothesis that these precursor cells serve as a reservoir for proliferating astrocytes and oligodendrocytes following injury. (2) To extend our studies of cellular localization of the glial-specific enzymes: Yp and Yb isoforms of glutathione-stransferase, carbonic anhydrase, glutamine synthetase and the RLM6 isozyme of cytochrome P-450 in the developing normal and myelin-deficient rat. These studies which will include immunostaining of tissue sections and cell cultures and enzyme assays of bulk isolated cells, are expected to help resolve some current issues in glial cell development, with particular emphasis on protoplasmic astrocytes. (3) To study the expression of a novel 66KD CNS-specific neurofilament protein in rat cerebellum during development and in cultured granule cells. The axonal cytoskeleton of the granule cells will be examined by electron microscopy and immuno-EM in vivo and in vitro. A cDNA probe will be constructed to the 66 KD NF protein and used for in situ hybridization in tissues and in Northern blot analyses of extracted RNAs. (4) To examine the heat-shock (stress) response in the CNS of Lewis rats induced to develop EAE, a disease involving inflammation, demyelination and extensive reactive gliosis; and in glial cells in culture during development and differentiation. Heat-stress mRNA and protein will be analyzed by Northern and Western blots, in situ hybridization, immuno-cytochemistry and in vitro protein synthesis. The methodology for these four studies includes cell isolation, cell culture, immunocytochemistry, enzyme assays, cDNA construction, gene sequencing, RNA isolation, Northern blots, Western blots, protein synthesis, gel electrophoresis, in situ hybridization, subcellular fractionation and polysome analysis. The long range goals of the program are to provide detailed information on some well defined processes of development and differentiation of the nervous system, so that we may better understand how they are perturbed by genetic or environmental influences. The results obtained should also lead to information concerning the generation of reactive astrocytes following injury and of oligodendro- cytes for remyelination.

该程序包含四个专门用于神经化学的项目， 神经系统发育的细胞生物学。 所有研究将是 在大鼠中进行。我们建议：（1）分离和纯化神经胶质前体 来自发育中和成熟大鼠大脑的细胞，跟随 这些细胞在培养中的分化，并检验假设 这些前体细胞是增殖的储层 受伤后的星形胶质细胞和少突胶质细胞。 （2）扩展我们的 神经胶质特异性酶的细胞定位：YP和YB 谷胱甘肽转移酶，碳酸酐酶，谷氨酰胺的同工型 发育中的细胞色素P-450的合成酶和RLM6同工酶 正常和髓磷脂缺乏大鼠。 这些研究将包括 组织切片，细胞培养物和酶测定的免疫染色 散装孤立的细胞有望帮助解决当前的一些问题 神经胶质细胞发育，特别强调原生质 星形胶质细胞。 （3）研究新型66KD CNS特异性的表达 大鼠小脑的神经丝蛋白在发育和 培养的颗粒细胞。 颗粒细胞的轴突细胞骨架 将通过电子显微镜和免疫EM在体内和IN进行检查 体外。 cDNA探针将被构造为66 kD NF蛋白，并使用 用于组织和北印迹分析的原位杂交 提取的RNA。 （4）检查中的热震（压力）响应 刘易斯大鼠的中枢神经系统诱发了EAE，这种疾病涉及 炎症，脱髓鞘和广泛的反应性神经病；和神经胶质 发育和分化过程中的培养细胞。热压力mRNA 和蛋白质将通过北部和蛋白质印迹分析，原位 杂交，免疫环化学和体外蛋白质合成。这 这四项研究的方法包括细胞分离，细胞培养， 免疫细胞化学，酶测定，cDNA构建，基因测序， RNA隔离，北印迹，蛋白质印迹，蛋白质合成，凝胶 电泳，原位杂交，亚细胞分级和 多层分析。该计划的远距离目标是提供 有关某些明确定义的开发过程和 神经系统的区分，以便我们可以更好地理解 它们如何受到遗传或环境影响的扰动。这 获得的结果也应导致有关 受伤后的反应性星形胶质细胞产生 细胞进行再髓。