microBSD:Spatiotemporal control of neurochemical tone in the brain slice using mi

描述（由申请人提供）：脑切片的制备提供了惊人的细胞和电路水平的大脑功能的细节。有许多类型的问题只能用脑切片来解决，但在实验中控制空间和时间的神经化学微环境是困难的，使用标准的水浴交换层流切片室。在我们的工作中，我们正在探索多巴胺（DA）和其他神经调节剂如何影响前额皮质微回路维持与工作记忆、认知灵活性和其他执行功能相关的时空活动模式的能力。多年的研究已经证明了DA功能障碍与认知功能的相关性——例如，在精神分裂症中，DA信号被认为是通过未知的机制受损，或者在帕金森病中，从腹侧被盖区（VTA）到皮层的DA神经支配丢失。我们希望能够在精确的时间和空间控制下，将神经化学物质（如DA）应用于同一切片的不同区域，以便以与完整大脑相关的方式探索对网络活动的影响。目前，我们仅限于在整个切片上交换浸泡介质，或者使用移液管将化合物吹入或吹入切片，这阻碍了电生理和成像的访问，并且无法很好地控制。对于我们和其他从事脑切片生理学研究的人来说，一个更精确、更通用的方法将是无价之宝。我们已经创建了一个原型微流控脑切片装置（5BSD），它将现成的脑切片室与设置在腔室底表面的微流控通道阵列结合在一起（http://www.jove.com/index/Details.stp?ID=302）。该设备使用快速原型制作，一旦优化，复制和与其他研究人员共享设备就很容易了。此外，我们的5BSD可以无缝集成到标准的生理/成像室中，可以立即提供给整个切片生理社区。有了这项技术，我们可以定位神经化学物质和任何其他可溶因素的流动，以我们选择的时间剖面精确定位大脑切片。我们对DA特别感兴趣，我们可以使用循环伏安法（CV）量化DA在组织中的递送。因此，我们将把DA输送到小鼠皮层有两个特定的目的——一个是进一步完善和发展这项技术，另一个是测试DA对皮层活动模式的影响。公共卫生相关性：我们的方法将提供一种新颖、复杂的方法来测试调节性神经递质及其拮抗剂对精神分裂症和帕金森病等多种疾病的神经回路动力学的影响。在控制神经化学微环境的同时直接成像其功能，很难想象有更好的方法来理解皮层微回路是如何工作的。我们设想，我们在这里的工作将导致一整类设备，这将使研究人员能够控制大脑切片准备的微环境，以解决各种问题，我们的努力整合到目前使用的腔室技术将为其他类似的努力树立一个标准，使生理学家能够轻松、立即获得

DESCRIPTION (provided by applicant): The brain slice preparation has provided amazing access to details of cellular and circuit level brain function. There are many classes of questions that can only be addressed using the brain slice, but controlling the spatial and temporal neurochemical microenvironment during experiments is difficult using standard bath exchange laminar flow slice chambers. In our work we are exploring how dopamine (DA) and other neuromodulators might influence the ability of the prefrontal cortical microcircuit to maintain spatiotemporal activity patterns associated with working memory, cognitive flexibility and other executive functions. Years of research have demonstrated the relevance of dysfunction of DA to cognitive function - for example in Schizophrenia where DA signaling is thought to be impaired through unknown mechanisms or in Parkinson's disease where DA innervation to cortex from the ventral tegmental area (VTA) is lost. We would like to be able to apply neurochemicals such as DA to different areas of the same slice under precise temporal and spatial control in order to explore effects on network activity in a way that is relevant to the intact brain. Currently, we are limited to exchanging the bathing medium over the whole slice or puffing compounds onto or into the slice using pipettes, which impede electrophysiological and imaging access and are not well controlled. A more precise and versatile method would be invaluable for us and for many others doing brain slice physiology. We have created a prototype microfluidic brain slice device (5BSD) that marries an off-the shelf brain slice chamber with an array of microfluidic channels set into the bottom surface of the chamber (http://www.jove.com/index/Details.stp?ID=302). This device is created using rapid prototyping and, once optimized, it is trivial to replicate and share the devices with other investigators. Additionally, our 5BSD integrates seamlessly into standard physiology/imaging chambers, it is immediately available to the whole slice physiology community. With this technology we can address the flow of neurochemicals and any other soluble factors to precise locations in the brain slice with the temporal profile we choose. We are interested specifically in DA and we can quantify DA delivery in tissue using cyclic voltammetry (CV). Therefore we will use DA delivery to mouse cortex for our 2 specific aims - one to refine and develop the technology further and the other to test the effects of DA on cortical activity patterns.PUBLIC HEALTH RELEVANCE: Our approach will offer a novel, sophisticated approach to test the effects of modulatory neurotransmitter and their antagonists on the circuit dynamics in diseases as diverse as schizophrenia and Parkinson's disease. It is difficult to imagine a better way of understanding how the cortical microcircuit works than by directly imaging its function while controlling the neurochemcial microenvironment. We envision that our work here will lead to a whole class of devices that will allow researchers to control the microenvironment of the brain slice preparation to address a variety of questions and that our efforts to integrate into currently used chamber technology will set a standard for other such efforts to be easily and immediately available for physiologists