NEUROBIOLOGY OF HEAT SHOCK RESPONSE IN NEURONAL CULTURES

我们最近证明谷氨酸的兴奋性毒性被减弱。 通过事先诱导热休克反应。一小段热度 应激诱导了一种蛋白质合成模式，这种模式具有 高度保守的热休克反应和增加HAR的表达 休克蛋白(HSP)基因。蛋白质的合成是必需的 神经保护作用。有证据表明，兴奋性毒性和 热休克反应在多种疾病中起着重要作用。 神经紊乱。许多细胞内和细胞间的过程具有 已被认为是体外兴奋性毒性的重要介体。这里 我们计划研究热休克反应的元素如何作用于 改变导致兴奋性毒性细胞死亡的事件顺序。在许多 在这些研究中，我们打算利用生长在 神经传递阻滞剂的存在。或之前的研究 这样的培养表明它们具有低基础水平的HSP表达和 当阻滞剂被移除时，它们表现出强大的癫痫样活动。 后一种特征我们计划作为突触介导的一种形式来研究 兴奋性应激，类似于体内的癫痫样活动。特指 我们建议对热休克蛋白的作用进行表征。 钙动态平衡，神经元的电生理反应 谷氨酸、谷氨酸激活磷脂酶A2和自发性 兴奋性神经传递。此外，使用最近可用的 免疫试剂我们计划仔细描述 热休克蛋白在大鼠神经元和神经胶质细胞中表达的神经保护模式 文化。预计这项工作将进一步加深我们对 热休克蛋白的保护特性及其重要机制 抑制给予神经保护。

We have recently demonstrated that glutamate excitotoxicity is attenuated by the prior induction of the heat-shock response. A short thermal stress induced a pattern of protein synthesis characteristic of the highly conserved heat-shock response and increase the expression of hear shock protein (HSP) mRNA. Protein synthesis was necessary for the neuroprotective effect. Evidence suggests that both excitotoxicity and the heat-shock response play a significant role in a variety of neurologic disorders. A number of intra-and intercellular processes have been proposed as important mediators of excitotoxicity in vitro. Here we plan to examine how the elements of the heat-shock response act to alter the sequence of events leading to excitotoxic cell death. In many of these studies we intend to utilized neuronal cultures grown in the presence of blockers of neurotransmission. Or previous studies with such cultures show that they have low basal levels of HSP expression and they exhibit robust, seizure-like activity when blockers are removed. The latter feature we plan to study as a form of synaptically mediated excitatory stress, akin to epileptiform activity in vivo. Specifically we proposed to characterized the effect of the heat-shock proteins on calcium homeostasis,electrophysiological responsiveness of neurons to glutamate, glutamate activation of phospholipase A2, and spontaneous excitatory neurotransmission. In addition, using recently available immunologic reagents we plan to carefully characterize the neuroprotective pattern of HSP expression in neurons and glial cells in the cultures. The work is expected to further our understanding of the protective properties of the HSPs and of the important mechanisms they inhibit to confer neuroprotection.