Synaptic transmission during status epilepticus

描述（由申请人提供）：一个为期五年的培训计划，为申请人提供发展所需的知识和技能，以进行未来的突触生理学和癫痫的基本机制的独立调查。在波士顿儿童医院完成了儿童神经病学和临床神经生理学的临床培训后，主要研究者现在是弗吉尼亚大学神经病学和儿科的助理教授。Jaideep Kapur，公认的癫痫基本机制的领导者，以及UVA神经病学系对这位候选人的承诺，使UVA成为申请人作为医生科学家培训的理想环境。拟议的研究将集中在提供一个更好的理解发生在癫痫持续状态抑制性突触的变化。用于治疗癫痫持续状态的第一线疗法（其靶向抑制性突触传递）通常失败，使患者处于死亡或显著神经系统发病的风险中。据推测，在癫痫持续状态期间抑制性突触发生的变化的更好的理解将提供一个框架，在此基础上的新疗法的目标是改善预后的癫痫持续状态。虽然两种不同的证据表明，抑制性突触传递的改变确实发生在癫痫持续状态期间，并且这种改变部分是突触处存在的GABAA受体补体的修饰的结果，但修饰的机制尚不清楚。使用癫痫持续状态的体外和体内模型结合电生理学和细胞成像技术，将通过完成四个特定目的来研究突触后GABAA受体群体中修饰的潜在机制：1）通过膜片钳记录来表征体外癫痫持续状态对GABA介导的突触传递的影响，2）通过膜片钳记录来表征体内癫痫持续状态对GABA介导的突触传递的影响，3）通过免疫细胞化学结合显微镜观察来表征癫痫持续状态对GABAA受体的内化率和分布的影响;采用膜片钳技术研究癫痫持续状态对突触前末梢GABA释放的影响。

DESCRIPTION (provided by applicant): A five-year training program providing the applicant with the opportunity to develop the knowledge and skills required to perform future independent investigations in synapse physiology and the basic mechanisms of epilepsy is proposed. Having completed clinical training in child neurology and clinical neurophysiology at Children's Hospital Boston, the principal investigator is now an Assistant Professor of Neurology and Pediatrics at the University of Virginia. The opportunity to expand upon his scientific skills under the mentorship of Dr. Jaideep Kapur, a recognized leader in the basic mechanisms of epilepsy, and the UVA Department of Neurology's commitment to this candidate makes UVA the ideal setting for the applicant's training as a physician-scientist. The proposed research will focus on providing an improved understanding of the changes that occur at inhibitory synapses during status epilepticus. First line therapies for the treatment of status epilepticus, which target inhibitory synaptic transmission, often fail leaving the patient at risk for mortality or significant neurological morbidity. It is posited that an improved understanding of the changes that occur at inhibitory synapses during status epilepticus will provide a framework on which to base new therapies with the goal of improving the prognosis for those who present in status epilepticus. While two distinct lines of evidence suggest that an alteration in inhibitory synaptic transmission does occur during status epilepticus and that this alteration is, in part, the result of a modification in the complement of GABAA receptors present at the synapse, the mechanism underlying the modification is not known. Using an in vitro and an in vivo model of status epilepticus combined with electrophysiological and cellular imaging techniques, the mechanism underlying the modification in the post-synaptic GABAA receptor population will be investigated by completing four Specific Aims: 1) To characterize the impact of in vitro status epilepticus on GABA-mediated synaptic transmission by means of patch clamp recording, 2) To characterize the impact of in vivo status epilepticus on GABA-mediated synaptic transmission by means of patch clamp recording, 3) To characterize the impact of status epilepticus on the rate of internalization and distribution of GABAA receptors by means of immunocytochemistry combined with microscopy and 4) To characterize the impact of status epilepticus on GABA release from the pre-synaptic terminals by means of patch clamp recording.