New Methodologies for Connectomics

项目摘要 动物的神经系统是由大量突触连接的神经元组成的。 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知识 在许多物种中都有信息。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，理解发展、组织、 而神经回路的中断是至关重要的。连接学的目标是生产和研究地图 神经系统内的神经元连接。连接研究需要自动成像 获取高倍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接线图的方法。但连接性地图本身就是 然而，这不足以解释大脑的功能。其他信息，如分子 神经元的身份，需要从相同的神经系统中提取。这样做的主要目标是 建议发展整个小鼠大脑和其他大型大脑样本的重金属染色 要获得完整连接体(Aim1)的体积电子显微镜图谱，请生成 用于相关光学和电子显微镜成像的小型化蛋白质结合剂桥接 连接与神经细胞类型研究(AIM2)，并扩大X射线显微镜在 多路脑成像(Aim3)。这些研究将为开发 连接学和建立多模式成像的新范式。 我的职业目标是经营一家学术实验室，开发大脑研究的基本方法。 集中于连接学最紧迫需求的拟议工作将成为支柱 为了我的独立研究，并允许我将新学到的知识整合到神经生物学中， 生物化学和显微镜，以及我在化学和材料科学方面的跨学科培训。 利希特曼实验室独特的环境将使我处于有利地位，以便实现我的 职业抱负。我和我的导师杰夫·利希特曼教授制定了一份详细的培训计划，以 帮助我过渡到独立。我会定期与他会面，讨论研究进展， 助学金撰写、学生辅导和实验室管理的策略。我将监督一个 研究生来练习我的导师技能。扩大我的科学关系网，建立 未来的协作，我将在研讨会、连接学会议、SFN和ACS中展示我的工作 年度会议。作为哈佛大学分子和细胞生物学系的一员，我将 有机会接触到神经科学、分子生物学、细胞生物学和生物化学的领军人物，以及 尖端核心设施。大脑计划多样性K99/R00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.