MOLECULAR MECHANISM OF RETINAL GANGLION CELL DEATH

描述（改编自申请人摘要）：视网膜神经节细胞死亡 在各种视神经疾病中，最常见的是 青光眼 研究人员和其他人进行的先前研究表明， 在实验性青光眼和其他视神经损伤模型中， 具有已知的程序性细胞死亡形式特征的神经疾病 细胞凋亡。 这种形式的细胞死亡是由基因控制的， 更好地了解调控这一过程的基因 将导致更好的治疗方法， 死亡的过程。 这一建议旨在确定三个机构的作用， 基因在神经节细胞死亡的调节。 这些基因p53 bcl-x 和bax形成了一个分子开关， 在调节多种细胞类型的凋亡的控制步骤。 早期 研究表明这些基因在神经节细胞中表达。 一组 本建议的具体目标是确定，使用组合 定量和定位研究，如果这些基因的表达， 神经节细胞以分子开关预测的方式改变 假设（例如，p53表达增加导致bcl-x减少 表达和bax表达的增加）。 其他具体目标 包括直接测试这些基因在神经节细胞中的功能 死亡过程。 这些直接测试将在基因上进行， 具有缺陷性p53或bax表达的改变的转基因小鼠， 这些实验的基本实验设计是 使用两种不同的方法刺激小鼠神经节细胞死亡（因此 这些基因的功能可以在不同的途径中进行测试， 神经节细胞死亡），包括视神经的部分挤压， 玻璃体内注射不同剂量的谷氨酸类似物 N-甲基-D-天冬氨酸，然后定量分析 细胞死亡

DESCRIPTION (Adapted from applicant's abstract): Retinal ganglion cells die in a variety of optic nerve diseases, the most prevalent of which is glaucoma. Previous studies conducted by the investigator and others showed that ganglion cells died in experimental glaucoma and other models of optic nerve disease with characteristics of a form of programmed cell death known as apoptosis. This form of cell death is genetically controlled and it is likely that a better understanding of the genes that regulate this process in ganglion cells will lead to better treatments that can be used to block the death process. This proposal is aimed at determining the role of three genes in the regulation of ganglion cell death. These genes, p53, bcl-x, and bax, appear to form a molecular switch that acts as one of the early control steps in regulating apoptosis in a variety of cell types. Early work has shown that these genes are expressed in ganglion cells. One set of specific aims in this proposal is to determine, using a combined quantitative and localization study, if the expression of these genes in ganglion cells is altered in a fashion predicted by the molecular switch hypothesis (e.g., that p53 expression increases causing a decrease in bcl-x expression and an increase in bax expression). The remaining specific aims involve direct tests of the functions of these genes in the ganglion cell death process. These direct tests will be carried out on genetically altered transgenic mice that have either defective p53 or bax expression or overexpress bcl-x, The basic experimental design of these experiments is to stimulate ganglion cell death in mice using two different approaches (thus the functions of these genes can be tested in diverse pathways leading to ganglion cell death), which include a partial crush of the optic nerve and intravitreal injection of varying doses of the glutamate analog N-methyl-D-aspartate, followed by a quantitative analysis of the rate of cell death.