NMDA Receptors and Ethanol Withdrawal Seizures

描述（由申请人提供）：目前的调查将专门关注NMDA受体（NMDAR）在触发电描记乙醇戒断癫痫发作中的作用，这是慢性乙醇滥用者发病率和死亡率的主要原因。 电生理和显微荧光监测将用于严格检查慢性乙醇暴露和戒断引起的NMDAR介导的神经传递的变化，以及这些变化与乙醇戒断过度兴奋发作之间的时间关系。使用海马盲片膜片钳制备，在长期乙醇给药的情况下以及在乙醇停药后的增量时间点，从海马外植体记录NMDAR介导的微型突触事件。 这些事件的频率和振幅分析将用于确定戒断诱导的电生理变化的突触前或突触后焦点。戒断诱导的NMDAR介导的微型事件的变化将与通过长期场电位监测检测到的癫痫样放电模式的发作在时间上相关。使用多光子激发以增强分辨率并限制光氧化，将采用GFP标记的NMDAR的共聚焦成像来直接可视化戒断诱导的NMDAR运输变化。

DESCRIPTION (provided by applicant): The present line of inquiry will focus exclusively on the role of NMDA receptors (NMDARs) in triggering electrographic ethanol withdrawal seizures, a prime cause of morbidity and mortality in chronic ethanol abusers. Electrophysiological and microfluorometric monitoring will be employed in the service of a rigorous examination of the changes in NMDAR-mediated neurotransmission caused by chronic ethanol exposure and withdrawal, and the temporal relationship between these changes and the onset of ethanol withdrawal hyperexcitability. Using the hippocampal blind slice patch-clamp preparation, recordings of NMDAR-mediated miniature synaptic events will be made from hippocampal explants in the setting of chronic ethanol administration, and at incremental time-points after withdrawal of ethanol. Frequency and amplitude analysis of these events will be employed to determine a pre- or postsynaptic focus of withdrawal-induced electrophysiological changes. Withdrawal-induced changes in NMDAR-mediated miniature events will be temporally correlated with the onset of epileptiform discharge patterns detected via long-term field potential monitoring. Confocal imaging of GFP-tagged NMDARs expressed by explanted hippocampal neurons, using multiphoton excitation to enhance resolution and limit photo-oxidation, will be employed to directly visualize withdrawal-induced changes in NMDAR trafficking.