New Methodologies for Connectomics

项目摘要 动物的神经系统由大量突触连接的神经元组成。 由此产生的神经网络是动物行为的基础，并有助于存储学习到的信息。 许多物种的信息。在人类中，神经网络的错误连接可能会导致 行为、学习和思想。由于所有这些原因，了解发展，组织， 神经回路的中断是至关重要的连接组学的目标是产生和研究 神经系统内的神经元连接。连接组学研究需要自动化图像 获取高放大率下覆盖大体积的脑组织图像，以解析突触 以及从这些图像生成布线图的方法。但连通图本身 但不足以解释大脑的功能。其他信息，如分子 神经元的身份，需要从同一个神经系统中提取。这个的主要目标 一项建议是开发整个小鼠大脑和其他大型大脑样本的重金属染色 对于完整连接体（Aim 1）的体积电子显微镜映射，产生以下文库： 用于相关光和电子显微镜成像的小型化蛋白质结合剂， 连接组学与神经元细胞类型研究（Aim 2），并扩大X射线显微镜在 多路脑成像（Aim 3）。这些研究将为发展 connectomics和建立新的范例多模态成像。 我的职业目标是管理一个学术实验室，为大脑研究开发基本方法。 拟议中的工作重点是连接组学最迫切的需求，将成为中流砥柱 我的独立研究，并允许我整合新获得的神经生物学知识， 生物化学和显微镜与我在化学和材料科学的跨学科培训。 Lichtman实验室独特的环境将使我处于一个特殊的位置，以追求我的 职业抱负。我和我的导师Jeff Lichtman教授一起制定了详细的培训计划， 帮我过渡到独立我将定期与他会面，讨论研究进展， 资助写作、学生指导和实验室管理的策略。我将监督一个 来练习我的指导技巧为了扩大我的科学网络， 未来的合作，我将在研讨会上介绍我的工作，连接会议，SfN和ACS 年度会议。作为哈佛分子和细胞生物学系的一员，我将 有机会接触神经科学，分子生物学，细胞生物学和生物化学的领导者，以及 尖端的核心设施。BRAIN倡议多样性K99/R 00将为我提供所需的资金 启动一项雄心勃勃的研究计划，以应对围绕大脑研究的突出挑战。

PROJECT ABSTRACT The nervous systems of animals are comprised of neurons connected by a large number of synapses. The resulting neural networks underlie animal behavior and contribute to the storage of learned information in many species. In humans, the miswiring of neural networks likely results in disorders of behavior, learning, and thought. For all these reasons, understanding the development, organization, and disruptions in neural circuits is vital. The goal of connectomics is to produce and study the maps of neuronal connections within nervous systems. Connectomic research requires automated image acquisition of brain tissue images that cover large volumes at high magnification to resolve synapses and methods to generate wiring diagrams from these images. But the connectivity map itself is nevertheless not sufficient to explain the brain functions. Additional information, such as the molecular identity of neurons, needs to be extracted from the same nervous system. The primary goal of this proposal is to develop the heavy metal staining of whole mouse brains and other large brain samples for volumetric electron microscopic mapping of a full connectome (Aim1), generate a library of miniaturized protein binders for correlated light and electron microscopic imaging to bridge connectomics with neuronal cell type studies (Aim2), and expand the use of X-ray microscopy in multiplexed brain imaging (Aim3). These studies will lay the foundation for the development of connectomics and establish new paradigms for multimodal imaging. My career goal is to run an academic lab that develops essential methodologies for brain research. The proposed work that focuses on the most urgent needs of connectomics will become a mainstay for my independent research and allow me to integrate the newly acquired knowledge in neurobiology, biochemistry, and microscopy with my interdisciplinary training in chemistry and materials science. The unique environment at Lichtman lab will put me in a privileged position in order to pursue my career aspirations. I have developed a detailed training plan with my mentor, Prof. Jeff Lichtman, to help me transition to independence. I will meet regularly with him to discuss research progress, strategies for grant writing, student mentorship, and lab management. I will oversee the work of a graduate student to practice my mentorship skills. To broaden my scientific network and establish future collaborations, I will present my work in workshops, connectomics meetings, SfN and ACS annual meetings. As a member of the Harvard Department of Molecular and Cellular Biology, I will have access to leaders in neuroscience, molecular biology, cell biology, and biochemistry, as well as cutting-edge core facilities. The BRAIN Initiative Diversity K99/R00 will provide me the funding required to initiate an ambitious research plan to tackle the outstanding challenges surrounding brain studies.