In Vivo Fluorescent Microscopy Analysis of Motor Cortex Activation by STN DBS

项目总结 丘脑底核(STN)深部脑刺激(DBS)是帕金森病的常用外科治疗方法 (Pd)。尽管STN DBS在临床上取得了20多年的成功，但其治疗机制仍然难以捉摸。 然而，很明显，DBS在分子、细胞和系统水平上发挥作用，在复杂和 有时是相互矛盾的方式。目前的技术，如电生理学、电化学和功能 通常用于研究这一谜题片段的成像在分辨率或行为范式方面都受到限制 这些可以应用于，因此没有提供解析复杂的 刺激和诱发效应之间的关系。在这里，我们建议使用荧光钙 用头戴式微型单光子系统作为新工具对表达GCaMP6f的大鼠进行显微镜观察 在清醒行为动物模型中弥合细胞和系统水平对DBS的理解差距 警察。为此，我们将分析刺激STN引起的运动皮质神经活动的变化。 开场、步进、圆柱体测试和阿朴吗啡诱导的旋转，所有这些都是刻板印象的测试， 已经显示了预测有效性，用于评估帕金森病动物的运动和治疗效果。这个 这里提出的技术为回答有关DBS机制的问题提供了一种独特的方法，例如 DBS对兴奋性STN谷氨酸能神经元的激活或药物抑制作用 投射到苍白球内侧/黑质网状质产生可检测到的运动变化 与行为结果变化相关的皮质活动(例如，开阔视野、运动学评估 步进法、圆柱法和阿朴吗啡诱导的旋转试验)。

PROJECT SUMMARY Subthalamic nucleus (STN) deep brain stimulation (DBS) is a common surgical treatment for Parkinson’s disease (PD). Despite over 20 years of clinical success, the therapeutic mechanisms of STN DBS remain elusive. However, it has become clear that DBS acts at the molecular, cellular, and systems levels in complex and sometimes contradictory ways. Current techniques such as electrophysiology, electrochemistry, and functional imaging commonly used to study pieces of this puzzle are limited in either resolution or behavioral paradigms these can be applied to, and have thus not provided all the information needed to parse out the complicated relationships between stimulation and evoked effects. Here, we propose the use of fluorescence calcium microscopy in GCaMP6f-expressing rats using a head-mounted miniature single photon system as a novel tool to bridge the gap between cellular and system level understanding of DBS in awake behaving animal models of PD. To this end, we will analyze neural activity changes in motor cortex evoked by stimulation of the STN during open field, stepping, cylinder tests, and apomorphine-induced rotations, all of which are stereotypical tests that have shown predictive validity for evaluation of movement and therapeutic efficacy in parkinsonian animals. The techniques proposed here provide a unique approach for answering questions about DBS mechanisms such as whether DBS-induced activation or pharmacologic inhibition of excitatory STN glutamatergic neuronal projections to the globus pallidus internus / substantia nigra reticulata produces detectable changes in motor cortex activity associated with changes in behavioral outcomes (e.g., open field, kinematic assessment of stepping, cylinder, and apomorphine-induced rotation tests).