Synaptic transmission during status epilepticus

描述（由申请人提供）：一个为期五年的培训计划，为申请人提供发展未来在突触生理学和癫痫基本机制方面进行独立研究所需的知识和技能的机会。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，首席研究员现在是弗吉尼亚大学神经病学和儿科学的助理教授。在癫痫基本机制领域公认的领导者Jaideep Kapur博士的指导下，他有机会扩展他的科学技能，并且弗吉尼亚大学神经内科对这位候选人的承诺使弗吉尼亚大学成为申请人作为一名医生科学家培训的理想场所。提出的研究将集中在提供一个更好的理解，发生在抑制性突触在癫痫持续状态。针对抑制性突触传递的治疗癫痫持续状态的一线疗法往往失败，使患者面临死亡或严重神经系统疾病的风险。我们认为，对癫痫持续状态期间抑制性突触发生的变化的更好理解将为新疗法的基础提供一个框架，以改善癫痫持续状态患者的预后。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，这种改变部分是突触中GABAA受体补体修饰的结果，但这种修饰的机制尚不清楚。结合电生理和细胞成像技术，通过体外和体内癫痫持续状态模型，研究突触后GABAA受体群体改变的机制，完成以下四个特定目的：1)通过膜片钳记录表征体外癫痫持续状态对gaba介导的突触传递的影响；2)通过膜片钳记录表征体内癫痫持续状态对gaba介导的突触传递的影响；3)采用免疫细胞化学结合显微镜观察观察癫痫持续状态对GABAA受体内化率和分布的影响；4)采用膜片钳记录方法观察癫痫持续状态对突触前末端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.