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

项目摘要 丘脑底核脑深部电刺激术是治疗帕金森病的一种常用手术方法 （PD）。尽管在临床上取得了20多年的成功，但DBS的治疗机制仍然难以捉摸。 然而，已经清楚的是，DBS在分子、细胞和系统水平上以复杂的方式起作用， 有时是矛盾的方式。目前的技术，如电生理学，电化学，和功能 通常用于研究这一难题的成像技术在分辨率或行为模式方面都有局限性 这些可以应用于，因此没有提供解析出复杂的 刺激和诱发效应之间的关系。在这里，我们建议使用荧光钙 使用头戴式微型单光子系统作为新工具在表达GCaMP 6 f的大鼠中进行显微镜检查 在清醒行为动物模型中，弥合DBS的细胞和系统水平理解之间的差距， 警局为此，我们将分析运动皮层神经活动的变化引起的刺激的延髓， 旷场、踏步、圆筒测试和阿扑吗啡诱导的旋转，所有这些都是常规测试， 已经显示出对帕金森病动物的运动和治疗效果的评估的预测有效性。的 这里提出的技术提供了一种独特的方法来回答有关DBS机制的问题， 无论是DBS诱导的兴奋性多巴胺能神经元的激活还是药理学抑制， 投射到苍白球内/黑质网状物产生可检测的运动变化， 与行为结果的变化相关的皮层活动（例如，开阔场地，运动学评估 步进、圆柱体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).