Synaptic transmission during status epilepticus

描述(由申请者提供)：一个为期五年的培训计划，为申请者提供机会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研究。在波士顿儿童医院完成了儿童神经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学和儿科的助理教授。在癫痫基本机制领域公认的领导者贾伊迪普·卡普尔博士的指导下，以及弗吉尼亚大学神经病学系对这位候选人的承诺，使弗吉尼亚大学成为申请者作为内科科学家进行培训的理想场所。这项拟议的研究将侧重于提供对癫痫持续状态期间抑制性突触发生变化的更好理解。针对抑制性突触传递的癫痫持续状态的一线治疗方法往往失败，使患者面临死亡或重大神经疾病的风险。有人认为，对癫痫持续状态期间抑制性突触变化的更好理解将为以改善癫痫持续状态患者预后为目标的新治疗提供一个框架。虽然有两条不同的证据表明，在癫痫持续状态期间，抑制性突触传递确实发生了变化，而且这种变化在一定程度上是突触中存在的GABAA受体补体发生变化的结果，但这种变化背后的机制尚不清楚。利用癫痫持续状态的体外和体内模型，结合电生理和细胞成像技术，通过完成四个具体目标来研究突触后GABAA受体群体的改变机制：1)通过膜片钳记录来表征体外癫痫状态对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状态对GABA介导的突触传递的影响；3)用免疫细胞化学结合显微镜技术研究癫痫持续状态对GABAA受体内化和分布的影响；4)用膜片钳技术研究癫痫持续状态对突触前终末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.