The role of thalamic T-type calcium channels in ethanol withdrawal

描述（由申请人提供）：本培训计划提案的重点是确定多次乙醇戒断后丘脑内发生的低阈值电压门控钙通道（T型）的细胞和分子变化。通过将这些变化与逐渐严重的戒断症状的发展联系起来，我希望了解乙醇戒断对丘脑功能的影响。因此，确定针对这些通道的治疗方法可能会为酒精中毒患者的成功康复提供新的有效的药物治疗。要解决这些问题，就需要在戒酒的技术和概念方法方面进行新的培训。我将通过研究三个综合目标来实现这一点。前两个目标将是确定T型钙通道如何有助于响应于多次乙醇暴露和戒断小鼠模型的潜在神经元变化。在目标1中，使用真实的时间RT-PCR，我将检查丘脑T型通道基因的表达水平在小鼠在多次乙醇暴露在蒸汽室。我假设在乙醇戒断过程中T型通道基因表达增加，并且这些增加的程度将与戒断次数直接相关。在目标2中，1将记录暴露于多次戒断的小鼠中线丘脑神经元的全细胞T型电流。我假设一个伴随的，功能相关的T型通道功能的增加，以及通道动力学的改变，这可能是乙醇戒断期间癫痫发作的易感性增加的基础。目的3将确定T型钙通道抑制剂乙琥胺和抗坏血酸在乙醇戒断过程中的作用。我将使用短暂的纺锤波事件作为神经元兴奋性的直接测量来监测皮质EEC活动，从而提供了戒断性癫痫发作易感性的间接测量。我假设已经给予乙琥胺和抗坏血酸的小鼠中纺锤体事件的量减少。酒精戒断综合征是一个严重的社会问题。这项研究计划很重要，因为它将为我提供必要的培训，以了解慢性乙醇暴露和随后的戒断反应中发生的长期分子和生理变化。目前用于酒精戒断的治疗方法，如苯二氮卓类药物，只能解决即时症状，并可以掩盖多次戒断期间发生的潜在生理变化，这些变化可能会引起更严重的症状。对酒精戒断综合征进展机制的更精确理解为确定新的辅助药物干预提供了巨大的机会，这些干预可以帮助成功治疗酒精戒断患者。

DESCRIPTION (provided by applicant): This training plan proposal is focused on determining the cellular and molecular alterations of low-threshold, voltage-gated calcium channels (T-type) that occur within the thalamus after multiple ethanol withdrawals. By linking these changes to the development of progressively more severe withdrawal symptoms, I hope to understand the effects of ethanol withdrawal on thalamic function. Identifying treatments that then target these channels may therefore present new and effective pharmacotherapies in the successful rehabilitation of alcoholic patients. To address these issues will require new training in technical and conceptual approaches to alcohol withdrawal. I will accomplish this by investigating three integrated aims. The first two aims will be to determine how T-type calcium channels contribute to underlying neuronal changes in response to a multiple ethanol exposure and withdrawal mouse model. In Aim 1, using real time RT-PCR, I will examine thalamic T-type channel gene expression levels in mice during and after multiple ethanol exposures in a vapor chamber. I hypothesize an increase in T-type channel gene expression during ethanol withdrawal, and that the degree of these increases will be directly related to the number of withdrawals. In Aim 2,1 will record whole cell T-type currents from midline thalamic neurons of mice exposed to multiple withdrawals. I hypothesize a concomitant, functionally relevant increase in T-type channel function, as well as alterations in channel kinetics which may underlie the increased susceptibility to seizures during ethanol withdrawal. Aim 3 will be to determine the effects of the T-type calcium channel inhibitors ethosuximide and ascorbate during ethanol withdrawal. I will monitor cortical EEC activity using brief spindle episodes as a direct measure of neuronal excitability, thus providing an indirect measure of withdrawal seizure susceptibility. I hypothesize a reduction in the amount spindle incidents in mice that have been administered ethosuximide and ascorbate. Alcohol withdrawal syndrome is a significant societal problem. This research proposal is important because it will provide me with the necessary training to understand the long-term molecular and physiological changes that occur in response to chronic ethanol exposure and subsequent withdrawal. Current treatments for alcohol withdrawal, such as benzodiazepines, only address the immediate symptoms and can mask underlying physiological changes occurring during multiple withdrawals that can give rise to more severe symptoms. A more precise understanding of the mechanisms involved in the progression of alcohol withdrawal syndrome offers a tremendous opportunity for identifying new adjunct pharmacological interventions that can aid in the successful treatment of alcohol withdrawal patients.