Optogenetic and chemogenetic manipulations of striatal output pathways for seizure control

项目摘要 深部脑刺激治疗癫痫的使用越来越多，并为那些需要其他治疗方法的患者提供了希望。 治疗失败。因此，识别用于刺激的新靶标具有相当大的临床和转化兴趣。 基底神经节（BG），已被广泛探索的刺激运动障碍，也可能 成为癫痫病的治疗目标临床前研究表明，调节BG核的活动，如 纹状体和黑质网状部（SNpr）可以有效地抑制急性模型的实验性癫痫发作 癫痫的症状然而，这种效应背后的网络机制，以及这种效应转化的程度， 慢性癫痫的模型仍然是未知的。 这个NRSA F30提案要求四年的支持。在第一年和第二年，我计划完成我的 博士论文本论文将包括以下研究计划中提出的两个目标， 大量更新，以反映我的研究进展，因为提交了原来的建议。 在目的1中，我将检验SNpr的按需光遗传学沉默将抑制SNpr基因的表达的假设。 慢性癫痫大鼠的自发性癫痫发作。与此同时，我将检验按需激活 会对自发性癫痫发作产生同等的抑制作用虽然这些操作是 在急性癫痫发作模型中进行了充分的研究，这些研究将是第一个评估 慢性癫痫的这些区域。在这个过程中，我将学习慢性癫痫持续状态模型， 在大鼠癫痫和建立独立的，自动化的系统，以监测其癫痫发作活动。 在目标2中，我将使用纤维光度法来检验癫痫活动进行性和 差异性地参与基底神经节输出通路。我还将检验光遗传学 对抗纹状体通路的病理性募集的操作将抑制癫痫发作。培训 这一目标包括学习纤维光度学的技术方面以及编码和数据分析 方法. 在我的博士研究过程中，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。我完成论文答辩后， 在奖学金期间的第3年和第4年完成我的博士前医学培训。临床培训如下 一个标准化的机构课程。经过这段时间的联合学位培训，我打算申请联合 在神经病学/癫痫学的医生-科学家培训计划中的住院医师/研究金轨道。 总之，在拟议的研究期间，我将获得尖端技术的专业知识， 神经生理学与智力和实际职业发展培训同时进行。工作提出的 将通过确定癫痫发作传播和通过基底神经节节点抑制的机制而使该领域受益 同时也测试它们作为慢性自发性癫痫管理的候选者。

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.