CELL DEATH AND THE REGULATION OF PURKINJE CELL NUMBER

细胞死亡与浦金野细胞数量的调节

• 批准号: 2873177
• 负责人: Michael W Vogel
• 金额: $ 13.69万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-02-01 至 2001-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2873177
• 关键词: apoptosis; cell growth regulation; cerebellar Purkinje cell; gene expression; genetically modified animals; laboratory mouse; neural degeneration; neurogenesis; protooncogene

DESCRIPTION: (Applicant's abstract) The focus of this study is on the mechanisms that regulate Purkinje cell number, both during normal development and in pathological conditions. Purkinje cells play a critical role in the regulation of pattern formation during cerebellar development. However, little is known about how Purkinje cell number is regulated. The consequences of Purkinje cell loss can also have profound effects on cerebellar development as in ataxia telangeictasia, a crippling inherited disorder of the immune and nervous systems. The experimental design of this study is to investigate normal and pathological programmed cell death in Purkinje cells using transgenic mice that overexpress a human bcl-2 gene (Hu-bcl-2) as experimental probes. Bcl-2 is a protooncogene that has been shown to protect neuronal and non-neuronal cells from programmed cell death. The initial studies of Hu-bcl-2 transgenic mice have shown that the number of Purkinje cells is increased by as much as 40%. This result suggests that overexpression of bcl-2 is rescuing Purkinje cells from naturally occurring cell death. The two specific aims of this grant proposal are 1) to investigate naturally occurring Purkinje cell death as a mechanism for regulating Purkinje cell number, and 2) to investigate the mechanisms of Purkinje cell loss in staggerer mutants. Naturally occurring Purkinje cell death will be investigated in three experimental protocols. First, the number of Purkinje cells in Hu-bcl-2 transgenic mice will be compared to controls during development to establish when Hu-bcl-2 expression begins to increase Purkinje cell number. Second, the expression of mouse and human bcl-2 will be examined in Hu-bcl-2 transgenics to determine if the transgene is expressed when Purkinje cell number is increased. Third, the cerebella of embryos and neonatal wild type mice will be examined for evidence of Purkinje cell degeneration using molecular probes for degenerating cells. The number of pyknotic Purkinje cells in wild type mice will be compared to Hu-bcl-2 transgenics. The mechanisms of Purkinje cell loss in staggerer mutants will be investigated by searching for evidence of Purkinje cell degeneration.

描述：（申请人摘要）本研究的重点是 调节浦肯野细胞数量的机制，在正常 发展和病理条件下。 浦肯野细胞在 在小脑发育过程中模式形成的调节作用。 然而，人们对浦肯野细胞数量如何调节知之甚少。 的 浦肯野细胞丧失的后果也可能对 小脑发育，如共济失调毛细血管扩张症，一种严重的遗传性 免疫系统和神经系统的紊乱。 这个实验设计 本研究旨在研究正常和病理性程序性细胞死亡， 使用过表达人bcl-2基因的转基因小鼠的浦肯野细胞 （Hu-bcl-2）作为实验探针。 Bcl-2是一种原癌基因， 显示保护神经元和非神经元细胞免于程序性细胞死亡。 Hu-bcl-2转基因小鼠的初步研究表明， 浦肯野细胞的数量增加了40%。 该结果表明 bcl-2的过度表达拯救了浦肯野细胞， 细胞死亡 这项拨款建议的两个具体目标是：1） 研究自然发生浦肯野细胞死亡作为 调节浦肯野细胞数量，2）研究 交错突变体的浦肯野细胞丢失。 自然产生的浦肯野细胞 死亡将在三个实验方案中进行研究。 一是 Hu-bcl-2转基因小鼠中的浦肯野细胞数量将与 在发育过程中控制，以确定Hu-bcl-2表达何时开始， 增加浦肯野细胞数量。 第二，小鼠和人的表达 将在Hu-bcl-2转基因中检查bcl-2，以确定转基因是否 当浦肯野细胞数量增加时表达。 第三，小脑 将检查胚胎和新生野生型小鼠的 浦肯野细胞变性的分子探针研究。 将野生型小鼠中的固缩浦肯野细胞的数量与对照小鼠中的固缩浦肯野细胞的数量进行比较。 Hu-bcl-2转基因。 癫痫大鼠浦肯野细胞丢失的机制 突变体将通过寻找浦肯野细胞的证据来研究 退化