Mechanisms of Alcohol Withdrawal

项目摘要 在理解酗酒如何导致酒精戒断综合症方面存在根本性的差距，其 一系列严重的临床症状，包括严重的癫痫发作。戒断发作呈进行性发作 每一次停药都会变得更糟，治疗会变得复杂，甚至可能是致命的。因此，至关重要的是要确定安全和 有效的治疗替代方案。钙通道可能为治疗提供了这样一种选择。长距离的 这项工作的目标是更好地了解酒精戒断的细胞和分子机制。 这项建议的近期目标是：1)确定T-型钙离子的细胞和分子变化 这些变化与癫痫的发生和发展有关。 酒精WD综合征的其他特征~3)以检查针对这些经络的治疗是否 有效地了解我们研究中涉及的T通道异构体的急性敏感性如何与 酒精WD综合征和癫痫涉及同一经络~和5)确定大脑如何相关 电路被招募到WD癫痫中。以强大的初步数据和公布的研究结果为指导，并由 作为一种研究WD癫痫的创新方法，将追求以下具体目标：目标1将确定 乙醇对T型通道的急性抑制如何导致WD过度兴奋。使用PKC的发现 调节乙醇的急性抑制效应，将使用全细胞记录来检验以下假设 在WD过程中，PKC抑制剂将揭示乙醇对T型电流的抑制作用的改变。目标2将 确定T型通道在多次酒精戒断所致癫痫发作中的作用。 定量脑电记录将用于测试T型通道阻滞剂是否会抑制WD发作和 将防止由于多个间歇性WDS而导致的癫痫的进展。AIM 3将确定选址 WD癫痫的来源和传播模式。WD发作的进展将使用 活体，多点记录，结合使用通道视紫红质2对这个网络的新颖的光遗传激活。 这些研究具有非常重要的意义，因为对T型钙的作用进行了垂直整合的研究 酒精戒断发作的通道，这是戒酒者治疗中的一个长期问题。 通道调制和网络参与的原理有望扩展到其他神经系统， 包括这些频道。这些研究具有创新性，因为它们提供了一种新的光遗传学方法 研究WD癫痫，这将被用来以前所未有的细节了解WD癫痫的进展， 以及一种基于当前抗癫痫药物的潜在新疗法。

Project Summary There are fundamental gaps in understanding how alcoholism leads to alcohol withdrawal syndrome, with its range of serious clinical symptoms, including profound seizures. Withdrawal seizures become progressively worse with each withdrawal, complicate treatment, and can be fatal. It is therefore vital to identify safe and effective treatment alternatives. Calcium channels may offer such an alternative for treatment. The long-range goal of this work is to better understand the cellular and molecular mechanisms underlying alcohol withdrawal. The immediate goals of this proposal are: 1) to define the cellular and molecular alterations in T-type Ca2+ channels that occur in response to multiple WDs~ 2) to link these changes to the development of seizure and other features of alcohol WD syndrome~ 3) to examine whether treatments targeting these channels are effective~ 4) to understand how the acute sensitivity of the T-channel isoform implicated in our studies relates to alcohol WD syndrome and seizures involving this same channel~ and 5) to determine how relevant brain circuitry is recruited into WD seizure. Guided by strong preliminary data and published findings, and enabled by an innovative method of studying WD seizures, the following specific aims will be pursued: Aim 1 will determine how acute inhibition of T-type channels by ethanol leads to WD hyperexcitability. Using the finding that PKC mediates the acute inhibitory effect of ethanol, whole cell recordings will be used to test the hypothesis that PKC inhibitors will reveal a change in ethanol-mediated inhibition of T-type currents during WD. Aim 2 will determine the contribution of T-type channels to seizures resulting from multiple ethanol withdrawals. Quantitative EEG recordings will be used to test whether T-type channel blockers will inhibit WD seizure and will protect against the progression of seizures due to multiple, intermittent WDs. Aim 3 will determine the site of origin and propagation patterns of WD seizure. The progression of WD seizure will be characterized using in vivo, multisite recordings coupled with novel optogenetic activation of this network using channelrhodopsin 2. The studies are highly significant because of the vertically integrated investigation of the role of T-type calcium channels in alcohol withdrawal seizure, which is an enduring problem in the treatment of abstaining alcoholics. The principles of channel modulation and network involvement promise to extend to other neural systems that include these channels. The studies are innovative because they deliver a new optogenetic approach to studying WD seizures, which will be used to understand the progression of WD seizure in unprecedented detail, and a potential new therapy based on a current antiepileptic drug.