Multifunctional Microprobe for Multiple Neurotransmitter Sensing and Optogenetics

描述（由申请人提供）：通过同时和近乎实时地测量大脑中多个神经活性分子的方法，将极大地促进理解网络内神经元如何相互作用以控制行为的努力。现有方法要么提供单一分析物的快速测量（例如，快速扫描循环伏安法），要么提供时间分辨率不足的多分析物测量（微透析）。在这里，我们将开发一种植入式微探针，能够同时快速监测3种普遍存在的神经递质/神经调节剂：多巴胺（DA），谷氨酸（Glut）和乙酰胆碱（ACh）。此外，我们将利用光遗传学的力量，将光波导集成到微探针中，为微电极阵列上的每个传感电极并置的组织提供定向照明。这将通过一种创新的硅基微探针设计来实现，该设计带有嵌入式波导和分光器，并在每个微电极下方嵌入一个微镜，以实现均匀的定向光照明。由此产生的微探针将结合基因隔离的神经元终端的局部调制，测量这些终端的递质释放，以及网络中神经元其他递质释放的最终变化，所有这些都在自由行为的动物的背景下进行。这些传感器将在广泛的神经科学应用中产生变革，因为DA、Glut和ACh作为神经元间信号分子参与多个大脑区域，控制从学习、记忆到运动控制的大量功能。我们对它们在体内表现的评估将集中在纹状体上，这是一个涉及运动控制和动机行为的大脑区域。多种递质释放事件与行为行为的精确耦合将为寻找多种神经和精神疾病的治疗干预措施提供有价值的信息。

DESCRIPTION (provided by applicant): Efforts to understand how neurons within networks interact to control behavior will be greatly facilitated by a means with which to measure multiple neuroactive molecules in the brain simultaneously and in near-real time. Existing methods either offer rapid measurements of a single analyte (e.g., fast-scan cyclic voltammetry) or provide multiple analyte measurement with insufficient temporal resolution (microdialysis). Here, we will develop an implantable microprobe capable of simultaneous rapid monitoring of 3 ubiquitous neurotransmitters/neuromodulators: dopamine (DA), glutamate (Glut), and acetylcholine (ACh). Further, we will harness the power of optogenetics by incorporating an optical waveguide into the microprobe providing directional illumination of tissue juxtaposed to each sensing electrode on the microelectrode array. This will be achieved using an innovative silica-based microprobe design with embedded waveguides and light splitters, and with a micromirror embedded underneath each microelectrode for uniform directional light illumination. The resultant microprobe will combine local modulation of genetically isolated neuron terminals with measurement of transmitter release from those terminals, together with resultant changes in the release of other transmitters from neurons within the network, all in the context of a freel behaving animal. These sensors will be transformative in a wide range of neuroscience applications due to the involvement of DA, Glut and ACh as inter-neuronal signaling molecules in multiple brain regions controlling a plethora of functions from learning and memory to motor control. Our evaluation of their performance in vivo will focus on the striatum, a brain region involved in motor control and motivated behavior. Precise coupling of multiple transmitter release events to behavioral actions will provide valuable information pertinent to the search for therapeutic interventions for multiple neurological and psychiatric disorders.