Optogenetic and chemogenetic manipulations of striatal output pathways for seizure control

项目摘要 深度脑刺激治疗癫痫的应用越来越多，并为其他疾病的患者提供了希望 治疗失败了。因此，确定新的刺激靶点具有相当大的临床和翻译意义。 基底节(BG)已被广泛用于运动障碍的刺激，它也可能 成为癫痫的诱人靶子。临床前研究表明，调节BG核的活动，如 纹状体和黑质网状部(SNPR)对实验性癫痫急性发作的抑制作用 癫痫的症状。然而，这种影响背后的网络机制，以及这种影响转化的程度 对慢性癫痫模型的研究仍不清楚。 此NRSA F30计划书需要四年的支持。在第一年和第二年，我计划完成我的 博士论文。本论文将围绕以下研究计划中提出的两个目标展开 大量更新，以反映我自提交原始提案以来的研究进展。 在目标1中，我将测试如下假设：SNPR的按需光遗传沉默将抑制 慢性癫痫大鼠的自发性癫痫发作。同时，我将测试按需激活的假设 对自发性癫痫发作有同等的抑制作用。虽然这些操纵是 在急性癫痫模型中得到了很好的研究，这些研究将是第一次评估阿司匹林的抗惊厥潜力。 这些区域在慢性癫痫中。在这个目标的过程中，我将学习慢性癫痫持续状态模型。 对大鼠的癫痫以及建立自给自足的自动化系统来监测它们的癫痫发作活动。 在目标2中，我将使用纤维光度法来检验癫痫发作活动递增和 不同地参与基底节的输出通路。我还将测试光遗传假说 对抗纹状体通路病理性募集的手法将抑制癫痫发作。培训 本课程的目的包括学习光纤光度测量的技术方面以及编码和数据分析 方法：研究方法。 在我的整个博士研究过程中，我将继续接受生理学和 组织学方法以及一般技能和职业发展。在我完成论文答辩后，我会 在奖学金期间的第三年和第四年完成我的医学博士前培训。接下来是临床培训 标准化的院校课程。在这段时间的联合学位培训后，我计划申请联合 住院医师/研究员跟踪神经学/癫痫学方面的医生-科学家培训计划。 总之，在拟议的奖学金期间，我将在以下领域获得尖端技术方面的专业知识 神经生理学与智力和实际职业发展方面的培训同步进行。建议的工作 将通过基底节节点确定癫痫的传播和抑制机制，从而使该领域受益 同时也测试它们作为治疗慢性自发性癫痫的候选药物。

Project Abstract Deep brain stimulation for epilepsy is growing in use, and offers promise to patients for whom other treatments fail. Thus, identifying new targets for stimulation is of considerable clinical and translational interest. The basal ganglia (BG), which have been explored extensively for stimulation in movement disorders, also may be attractive targets for epilepsy. Preclincial studies show that modulating activity in BG nuclei such as the striatum and substantia nigra pars reticulata (SNpr) can potently suppress experimental seizures in acute models of epilepsy. However, the network mechanisms underlying this effect, and the degree to which this translates into models of chronic epilepsy remain unknown. This NRSA F30 proposal requests four years of support. During years 1 and 2, I plan to complete my doctoral thesis. The thesis will encompass the two aims presented in the following research plan that have been heavily updated to reflect my research progress since the submission of the original proposal. In aim 1, I will test the hypothesis that on-demand optogenetic silencing of the SNpr will suppress spontaneous seizures in chronically epileptic rats. In parallel, I will test the hypothesis that on-demand activation of the striatum will produce an equivalent suppression of spontaneous seizures. While these manipulations are well-studied in acute seizure models, these studies would be the first to evaluate the anticonvulsant potential of these regions in chronic epilepsy. During the course of this aim, I will learn the status epilepticus model of chronic epilepsy in rats and the set up of self-contained, automated systems to monitor their seizure activity. In aim 2, I will use fiber photometry to test the hypothesis that seizure activity progressively and differentially engages basal ganglia output pathways. I will also test the hypothesis that optogenetic manipulations that counteract pathological recruitment of striatal pathways will suppress seizures. The training in this aim includes learning the technical aspects of fiber photometry as well as coding and data analysis methods. Throughout the course of my doctoral research, I will continue to be trained in physiological and histological methods as well as general skills and career development. After I complete my thesis defense, I will complete my pre-doctoral medical training during years 3 and 4 of the fellowship period. Clinical training follows a standardized institutional curriculum. After this period of joint-degree training, I plan to apply to combined residency/fellowship tracks in physician-scientist training programs in neurology/epileptology. In sum, during the proposed fellowship period, I will gain expertise in cutting-edge techniques in neurophysiology concurrent with training in intellectual and practical career development. The work proposed will benefit the field by determining mechanisms of seizure propagation and restraint via basal ganglia nodes while also testing them as candidates for the management of chronic spontaneous epilepsies.