MOLECULAR MECHANISM OF RETINAL GANGLION CELL DEATH

描述(摘自申请者摘要)：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。之前由调查员和其他人进行的研究表明 神经节细胞在实验性青光眼和其他视神经模型中死亡 以已知的一种程序性细胞死亡为特征的神经病 也就是细胞凋亡。这种形式的细胞死亡是由基因控制的，它是 很可能对调控这一过程的基因有更好的理解 将导致更好的治疗方法，可以用来阻断 死亡的过程。这项建议旨在确定三个方面的作用 调控神经节细胞死亡的基因。这些基因，P53，BCL-x， 和Bax，似乎形成了一个分子开关，起到了早期 调控多种细胞类型中的细胞凋亡的步骤。早些时候 研究表明，这些基因在神经节细胞中表达。一套 这项建议的具体目标是确定，使用组合 定量和定位研究，如果这些基因在 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bclx减少 表达和Bax表达增加)。剩下的具体目标 直接测试这些基因在神经节细胞中的功能 死亡过程。这些直接检测将在基因上进行 P53或Bax表达缺陷的转基因小鼠或 过表达bclx，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的神经节细胞死亡(因此 这些基因的功能可以在不同的途径上进行测试，从而导致 神经节细胞死亡)，包括视神经部分挤压和 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.