NITRIC OXIDE AS A MEDIATOR OF NEUROTOXICITY

在原代神经元培养中，N-甲基-D-天冬氨酸的神经毒性是 部分由一氧化氮(NO)介导。NO的作用机制 神经毒性以及NO的来源尚不清楚。因此， 将进行和设计实验，以进一步阐明 神经毒性方面没有。 选择性去除或灭活一氧化氮合成酶(NADPH)的条件 黄递酶)神经元将会发育。在此之后将详细介绍 这些培养物中兴奋性氨基酸神经毒性的分析。会的 确定兴奋后是否形成诱导型一氧化氮合酶 氨基酸的管理，以及它是否在NMDA中发挥作用 神经毒性。 已知一氧化氮合酶(NADPH黄递酶)神经元具有相对抗性 对NMDA神经毒性的影响。将进行实验以确定是否 这种保护不涉及合成酶。瞬变的条件 一氧化氮合酶在神经元中的表达将发展和兴奋性氨基 酸性神经毒性将进行详细研究。此外，这个角色， 抗氧化酶，如铜/锌超氧化物歧化酶，锰- 超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶和谷胱甘肽 还原酶在神经毒性和神经保护中的作用将被研究。 超氧阴离子在介导NO细胞死亡中的作用将是 通过探索NMDA神经毒性进行调查，以及外源性 在各种超氧化物歧化酶抑制剂存在的情况下应用NO 以及超氧阴离子的解放者。此外，细胞系超过和 低表达的锰超氧化物歧化酶将被开发出来。 利用这些细胞系，对外源应用NO及其受体的研究 随后的毒性将被调查。 最后，磷酸化或磷酸化的功能后果 将在原代神经元中研究一氧化氮合酶的去磷酸化 兴奋性氨基酸注射后培养。此外, 一氧化氮谷氨酸受体亚型(S)的测定 合成酶的激活将通过共转染研究来确定。

In primary neuronal cultures, N-methyl-D-aspartate neurotoxicity is mediated in part by nitric oxide (NO). The mechanisms involved in NO neurotoxicity, as well as the source of NO is not known. Accordingly, experiments will be performed and designed to further elucidate the role of NO in neurotoxicity. Conditions for the selective removal or inactivation of NO synthase (NADPH diaphorase) neurons will be developed. this will be followed by a detailed analysis of excitatory amino acid neurotoxicity in these cultures. It will be determined whether an inducible NO synthase is formed after excitatory amino acid administration, and whether it plays a role in NMDA neurotoxicity. NO synthase (NADPH diaphorase) neurons are known to be relatively resistant to NMDA neurotoxicity. Experiments will be performed to determine whether NO synthase is involved in this protection. Conditions for transient expression of NO synthase in neurons will be developed and excitatory amino acid neurotoxicity will be studied in detail. In addition, the role that antioxidant enzymes, such as copper/zinc-superoxide dismutase, manganese- superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase play in neurotoxicity and in neuroprotection will be studied. The role of the superoxide anion in mediating NO cell death will be investigated by exploring NMDA neurotoxicity, as well as exogenously applied NO in the presence of various inhibitors of superoxide dismutase and liberators of the superoxide anion. Furthermore, cell lines over and under-expressing manganese-superoxide dismutase will be developed. Employing these cell lines, studies on exogenously applied NO and its subsequent toxicity will be investigated. Finally, the functional consequences of phosphorylation or dephosphorylation of NO synthase will be investigated in primary neuronal cultures after excitatory amino acid administration. In addition, determination of the subtype(s) of glutamate receptor responsible for NO synthase activation will be identified by co-transfection studies.