PROGRAMMED CELL DEATH IN THE NERVOUS SYSTEM

我们研究了神经系统中的程序性细胞死亡和 一般的细胞凋亡的生化机制。接近紧张的人 系统，需要更灵敏和更原位的方法来鉴定细胞 正在经历细胞程序性死亡。我们开发了两种新方法来 在显微镜下鉴定凋亡细胞。(一)我们发现 胸腺细胞程序性细胞死亡在形态上可以发生 Nomarski光学和胸腺细胞死亡类似神经细胞 死亡。原子核解体的形态分析使我们能够 以测试细胞死亡是由于有毒因素的产生还是由于 失去了一种保护因素。使用新的微观方法 鉴定细胞凋亡，异核体中的核被发现遵循 亲代细胞的原始而独特的命运，而不是转移 细胞核之间的凋亡或存活率。这种新方法也能让我们 细胞凋亡是小脑颗粒细胞死亡的一种方式 经MPP+处理后，体外培养。(2)我们还开发了一种分子 原位检测DNA链断裂的方法。这使我们能够 检查正在经历神经退行性变化的动物的大脑 缺血、MPTP治疗和发育过程中。这种新方法有 使我们能够确定细胞凋亡在发育和发育过程中扮演的角色 在神经系统的各种疾病状态下。3)我们已经开发出 抗凋亡调控基因Bcl2、Bclx和Bcl2的单抗 探讨BAX在体内外对细胞凋亡的调节作用。

We have studied programmed cell death in the nervous system and the biochemical mechanism of apoptosis in general. To approach the nervous system, more sensitive and in situ methods are needed to identify cells undergoing programmed cell death. We have developed two new methods to identify apoptotic cells under the microscope. (1) We have found that thymocyte-programmed cell death can be followed morphologically with Nomarski optics and that the thymocyte death resembles neuronal cell death. The morphologic analysis of nuclear disintegration has allowed us to test whether cell death is due to production of a toxic factor or due to the loss of a protective factor. Using the new microscopic method to identify apoptosis, the nuclei in the heterokaryons were found to follow the original and distinct fate of the parent cells and not to transfer apoptosis nor viability between nuclei. This new method also allowed us to identify apoptosis as the method of cerebellar granule cell death after MPP + treatment in vitro. (2) We have also developed a molecular detection method to measure DNA strand breaks in situ. This allows us to examine brains of animals undergoing neurodegenerative changes during ischemia, MPTP treatment, and during development. This new method has allowed us to determine the role apoptosis plays during development and during various disease states of the nervous system. 3) We have developed monoclonal antibodies against apoptosis regulatory genes Bcl-2, Bcl-x and Bax to probe the regulation of apoptosis in vitro and in vivo.