Mechanisms of MAP2 degradation after in vitro ischemia.

体外缺血后 MAP2 降解的机制。

• 批准号: 6412379
• 负责人: KATHLEEN M SUSMAN
• 金额: $ 16.36万
• 依托单位: VASSAR COLLEGE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-01-01 至 2003-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6412379
• 关键词: NMDA receptors; brain injury; calcineurin; calcium; calcium channel blockers; cerebral ischemia /hypoxia; confocal scanning microscopy; cytoskeletal proteins; glutamate receptor; hippocampus; immunocytochemistry; laboratory rat; mass spectrometry; matrix assisted laser desorption ionization; microtubule associated protein; neural degeneration; neurons; phosphorylation; protein degradation; stroke; tissue /cell culture; western blottings

DESCRIPTION (Adapted from applicant's abstract): The objective of this application is to elucidate the early cellular triggers of MAP2 degradation following in vitro ischemia in the rat hippocampal slice. MAP2 is a pivotal cytoskeletal protein that is degraded rapidly after ischemia and may play a key role in the irreversible morphological damage in neurons, leading ultimately to neurodegeneration. The earliest timepoint of MAP2 degradation will be determined and the spatiai location within the neurons where this occurs will be discerned. The role of NNDA and inetabotropic glutamate receptors and the role of calcium will be examined as possible triggers of the MAP2 breakdown. The source of damaging calcium will be identified using confocal laser scanning microscopy and calcium pathway blockers. The roles of calcineurin and calpain as the enzymes responsible for the MAP2 breakdown will be elucidated by measuring MAP2 levels using immunoblotting techniques and calpain inhibitors. The precise location of altered phosphorylation on MAP2 will be identified using MALDI-TOF mass spectrometry and the role of the phosphatase calcineurin in that dephosphorylation will be assayed using specific inhibitors of this enzyme. The results from this project will provide important new information about the mechanism of damage to this key cytoskeletal protein and will further the goal to understand how damage to the cytoskeleton is involved in cell death after stroke.

描述（改编自申请人摘要）：本研究的目的 应用是阐明MAP 2降解的早期细胞触发因素 在大鼠海马切片中体外缺血后。MAP 2是一个关键的 一种在缺血后迅速降解的细胞骨架蛋白， 在神经元不可逆的形态学损伤中的作用，最终导致 神经变性MAP 2降解的最早时间点为 确定，并且发生这种情况的神经元内的空间位置将 被识破NNDA和非代谢型谷氨酸受体的作用以及 钙的作用将作为MAP 2分解的可能触发因素进行检查。 使用共聚焦激光扫描将确定破坏性钙的来源 显微镜检查和钙通道阻断剂。钙调神经磷酸酶和钙蛋白酶的作用 因为负责MAP 2分解的酶将通过以下方式阐明： 使用免疫印迹技术和钙蛋白酶抑制剂测量MAP 2水平。 将确定MAP 2上改变的磷酸化的精确位置 使用MALDI-TOF质谱和磷酸酶钙调神经磷酸酶的作用 因为去磷酸化将使用该酶的特异性抑制剂进行测定， 酵素该项目的结果将提供重要的新信息 关于这种关键细胞骨架蛋白的损伤机制， 目的是了解细胞骨架的损伤如何参与细胞死亡 中风后