Functional Neuroimaging in Awake, Behaving Animals

描述（由申请人提供）：该提案提出了两个学术机构（约翰霍普金斯大学和肯尼迪克里格研究所）的实验室之间的合作，具有互补的能力和资源，以提供最先进的技术，用于在清醒的行为动物中同时光遗传学控制神经元活动和功能成像。 我们建议在两年内提供一个头戴式，多功能，无系绳的光学系统-一种新型的显微镜-能够远程控制基因靶向神经元和成像功能血流反应在自由移动的大鼠。 该提案的高度创新方面是用于光遗传学控制的光传递的小型化头戴式光学显微镜和用于使用激光散斑对比成像（LSCI）方法进行功能成像的独特的定制超大规模集成（VLSI）电路芯片。 本研究将通过对大鼠胡须参与纹理辨别的行为学研究，探讨感兴趣的桶状皮质区的失活如何影响脑血流的功能反应，以及它如何转化为外显行为。 为这一修订后的应用程序所做的初步工作已经导致了一个原型的大小和重量兼容的安装在清醒和未被吓倒的啮齿动物的显微镜，以及光学生产伪影免费的图像质量相当的台式系统。 由于盐视紫红质激活的光刺激导致的血流变化也首次被成像。 这项工作的高度影响源于在清醒和行为动物中进行功能神经科学研究的潜力，从而解决了需要约束和麻醉的最大限制之一。 这项技术将为操纵神经元回路，神经血管耦合，发育过程中的功能和结构变化，自然行为过程中的学习和可塑性以及中风，偏头痛或癫痫发作模型的新基础研究打开大门。

DESCRIPTION (provided by applicant): This proposal presents a collaboration between laboratories in two academic institutions (Johns Hopkins University and Kennedy Krieger Institute) with complementary capabilities and resources to deliver a state of the art technology for simultaneous optogenetic control of neuronal activity and functional imaging in awake, behaving animals. We propose to deliver within two years a head-mountable, multifunction, tether-free optical system - a novel microscope - capable of remotely controlling genetically targeted neurons and imaging functional blood flow response in free-moving rats. Highly innovative aspects of this proposal are the miniaturized head-mountable optical microscope for light delivery for optogenetic control and a unique, customized very large scale integrated (VLSI) circuit chip for functional imaging using the method of laser speckle contrast imaging (LSCI). A behavioral study on rat whisker involvement in texture discrimination will be carried out to explore how inactivation of the interested barrel cortex fields affects functional responses of cerebral blood flow and how it is translates into explicit behavior. Preliminary work done for this revised application has resulted in a prototype with size and weight compatible for mounting the microscope on awake and undeterred rodents, as well as optics producing artifact free image quality comparable to bench top systems. Changes in blood flow due to optical stimulation by halorhodopsin activation have also been imaged for the first time. The high impact of this work stems from the potential to do functional neuroscience research in awake and behaving animals and thereby addressing one of the biggest limitations of requiring restraint and anesthesia. This technology will open the door for new fundamental research on manipulating neuronal circuits, neurovascular coupling, functional and structural changes during development, and learning and plasticity during natural behavior as well as in models of stroke, migraine or seizure.