Creating an integrated platform for in vivo brain targeting and experimentation

中枢神经系统（CNS）控制着无数的生理过程，从存储记忆到控制肌肉运动和调节食欲。为了了解离散神经群体在体内的作用，现代神经科学已经开发出基于病毒的工具，用于用光控制神经元活动（光遗传学），并通过监测细胞内钙报告分子的荧光来量化神经元活动的实时变化（光度法）。这两个工具是神经科学家工具包的关键组成部分，可以破译动物行为背后的精确神经回路，但需要专门的复杂仪器。此外，光遗传学和光纤光度测定法的使用是基于以微米内的高空间精度和再现性将病毒载体递送至离散CNS神经元群体以表达光门控离子通道或钙报告基因的能力。为了促进这些病毒注射，使用立体定位框架的颅内手术是必不可少的，允许操纵臂在三维空间中以微米精度移动，以将病毒递送到大脑。这三种技术（高精度靶向、光度学和光遗传学）共同提供了广泛的神经科学研究途径;使我们能够跟踪和控制定义的CNS部位中包含的神经元，并进行功能研究，以了解它们在活体动物中的功能。该提案旨在为体内脑靶向和实验创建一个集成平台。我们申请资金购买两个高精度立体定位框架与显微注射器，光遗传设备的神经激活和光度测定设备在体内钙成像。至关重要的是，这台设备将缓解我们目前实验装置中的严重瓶颈，并对我们目前的硬件进行重大技术改进。重要的是，这种仪器将使七个NSERC支持的研究小组能够在麦吉尔大学健康中心研究所（RIMUHC）（Sabatini，Huang，Suvrathan，Stellwagen，Sjöström，Chen和Murai）进行有影响力的神经科学研究，并使麦吉尔大学更广泛的神经科学界在实施创新技术和方法方面受益匪浅。这些设备将在未来5年内用于培训50多名HQP，这些专业知识将使他们在科学，医学和工业等不同领域的职业生涯中受益匪浅。我们将专门利用这些设备为来自代表性不足的群体和发展中国家的学生举办培训和讲习班，我们集团每年定期主办这些培训和讲习班。由于我们的团队无法通过其他方式获得这种设备，因此该提案的成功资助将迅速增强我们NSERC资助研究的影响力，并允许使用强大的技术进行关键访问和培训，以了解行为背后的神经回路。

The central nervous system (CNS) controls myriad physiologic processes, from storing memories to controlling muscle movement and regulating appetite. To understand the roles of discrete neural populations in vivo, modern neuroscience has developed viral-based tools to control neuron activity with light (optogenetics) and to quantify real-time changes in neuronal activity by monitoring fluorescence of intracellular calcium reporters (photometry). These two tools are critical components of a neuroscientist's toolkit to decipher the precise neural circuits that underlie animal behavior, but require dedicated complex instrumentation. Additionally, the use of optogenetics and fiber photometry is predicated on the ability to deliver viral vectors with high spatial precision and reproducibility within micrometres to discrete CNS neuronal populations to express the light-gated ion channels or the calcium reporters. To facilitate these viral injections, intracranial surgeries using a stereotaxic frame are essential, allowing micrometre precision of movement in 3-dimensional space of a manipulator arm for viral delivery to the brain. Together, these three technologies (high precision targeting, photometry, and optogenetics) a broad range of neuroscience research avenues; allowing us to track and control neurons contained in defined CNS sites and perform functional studies to understand their function in the living animal. This proposal seeks to create an integrated platform for in vivo brain targeting and experimentation. We request funding to purchase two high-precision stereotaxic frames with microinjectors, optogenetic equipment for neuroactivation and photometry equipment for in vivo calcium imaging. Critically, this equipment will alleviate severe bottlenecks in our current experimental set-up and provide a major technological improvement over our current hardware. Importantly, this instrumentation will empower seven NSERC-supported research groups to pursue impactful neuroscience research at the Research Institute of the McGill University Health Centre (RIMUHC) (Sabatini, Huang, Suvrathan, Stellwagen, Sjöström, Chen, and Murai) and greatly benefit the broader neuroscience community at McGill University in implementing innovative techniques and approaches. This equipment will be used to train more than 50 HQP over the coming 5 years and this expertise will greatly benefit them as they pursue careers in diverse fields of science, medicine, and industry. We will specifically utilize this equipment in training and workshops for students coming from underrepresented groups and developing countries, which our group regularly hosts annually. As our team does not have access to this equipment through other means, the successful funding of this proposal will rapidly enhance the impact of our NSERC-funded research and allow critical access to and training with powerful technology to understand the neural circuits underlying behavior.