Potassium Channel Gene Therapy to Prevent Neuron Death

描述(申请者逐字？S摘要)：尽管基因治疗已经 对于大脑中的兴奋性毒性侮辱来说，这是一种很有前途的策略， 目前使用的载体不能很快表达功能蛋白 侮辱靶点最早的事件阅读到坏死性神经元损伤。这 提案测试了一种新的策略：潜在的结构性表达 一种保护性蛋白质，其活性会被侮辱所激活。癫痫发作是， 根据定义，兴奋毒性，减少这种兴奋可能是 具有神经保护作用。这可能是通过超极化兴奋神经元来实现的。 通过向外整流钾(K+)通道，从而延长恢复时间 动作电位的时相和/或缩短尖峰持续时间。保护者 我们将探索钙调节或电压门控K+通道的作用。在……里面 我们的初步数据表明，在原代培养的海马区，两者都具有保护作用 对抗谷氨酸、星孢菌素和氰化钠。我们将延长这些措施 通过测试更多的侮辱以及探索 保护的目标是坏死而不是凋亡。体内研究不仅将 检查是否存在神经元数量和功能的节余 (使用电生理和行为端点)。最后，我们将检查 这些通道在健康神经元中的活动，如果它们是活跃的，测试 它们对正常学习和记忆的影响。

DESCRIPTION (Verbatim from applicant?s abstract): Although gene therapy been shown to be a promising strategy for excitotoxic insults in the brain, currently used vectors do not express functional protein soon enough after insult to target the earliest events reading to necrotic neuronal injury. This proposal tests a novel strategy: constitutive expression of a potentially protective protein whose activity would be activated by insult. Seizures are, by definition, excitotoxic, and decreasing such excitation might be neuroprotective. This might be accomplished by hyperpolarizing excited neurons via outwardly rectifying potassium (K+) channels, thus prolonging the recovery phase of the action potential and/or shortening spike duration. The protective effects of a Ca2+-regulated or a voltage-gated K+ channel will be explored. In pilot data we have shown that both protect in primary hippocampal cultures against glutamate, staurosporine, and sodium cyanide. We will extend these findings by testing more insults as well as exploring the extent to which protection targets necrosis vs. apoptosis. In vivo studies will not only examine whether there is sparing of neuron number, but of function as well (using electrophysiological and behavioral endpoints). Finally, we will examine the activity of these channels in healthy neurons and, if they are active, test their effects on normal learning and memory.