PHYSIOLOGY OF APOPTOTIC NEURONAL INJURY IN VITRO

体外凋亡性神经元损伤的生理学

基本信息

批准号：
6393970
负责人：
Steven Mark Rothman
金额：
$ 22.78万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-09-01 至 2002-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6393970
关键词：
apoptosis autonomic nervous system confocal scanning microscopy cysteine endopeptidases enzyme activity granule cell human tissue laboratory rat nerve injury neurophysiology neurotrophic factors protease inhibitor tissue /cell culture voltage /patch clamp voltage gated channel

项目摘要

DESCRIPTION Over the past two decades biologists have recognized that there are two broad forms of cell death--necrosis and apoptosis. The former is seen after many severe, acute injuries and is associated with rapid cellular destruction. The latter can be triggered by a variety of acute and chronic insults and may take hours to weeks to fully manifest. There is now compelling evidence that a major component of the neuronal death seen in a variety of acute and chronic neurological diseases, including stroke, is apoptotic, and that this type of death is mediated by several endogenous cysteine proteases. The focus of this application is to determine the physiological consequences of aborting apoptotic neuronal death and to determine whether the triggers for apoptotic injury are present in human neocortex deprived of oxygen and glucose. Specifically: (1) Examine the function of voltage-gated and ligand-gated channels in autonomic ganglion neurons and cerebellar granule neurons that have been deprived of trophic factors but rescued from cell death by inhibition of proteases. (2) Determine the time course of protease activation in brain slices from rodents, and then humans (obtained from neocortical biopsy), and then identify interventions that inhibit protease activation. These experiments will utilize standard methods for intracellular recording and voltage clamping of cultured neurons and quantitative confocal imaging of brain slices. They will provide the first direct information about the physiological properties of neurons that have had an apoptotic death program interrupted. They should allow us to determine the functional potential of neurons when apoptosis has either been stabilized or actually reversed. Most important, they will provide direct evidence for the recruitment of apoptotic pathways in ischemic human brain.

在过去的二十年中，生物学家已经认识到那里是细胞死亡的两种广泛形式 - 衰老和凋亡。前者是在严重严重，急性损伤后看到，并与快速细胞有关破坏。后者可以由多种急性和慢性触发侮辱，可能需要数小时到几周才能充分体现。现在有令人信服的证据表明，神经元死亡的主要组成部分包括中风在内的急性和慢性神经系统疾病的种类繁多凋亡，这种类型的死亡是由几个内源性介导的半胱氨酸蛋白酶。该应用程序的重点是确定流产凋亡神经元死亡的生理后果确定人类中是否存在凋亡损伤的触发因素新皮层被剥夺了氧气和葡萄糖。具体：（1）检查自主神经节中电压门控和配体门控通道的功能神经元和小脑颗粒神经元被剥夺了营养因素，但由于抑制蛋白酶而从细胞死亡中救出。（2）确定大脑切片中蛋白酶激活的时间过程啮齿动物，然后是人类（从新皮质活检获得），然后确定抑制蛋白酶激活的干预措施。这些实验将利用标准方法用于细胞内记录和电压培养神经元的夹紧和大脑的定量共聚焦成像切片。他们将提供有关的第一个直接信息具有凋亡死亡程序的神经元的生理特性中断。他们应该允许我们确定神经元当凋亡稳定或实际逆转时。最重要的是，他们将为招募的直接证据提供缺血性人脑中的凋亡途径。