The basic wiring diagram of the brain: a global mouse brain connectome Phase II

描述（由申请人提供）：该提案是我们努力创建C57 Black/6 J小鼠大脑的三维、类似Google Earth的数字连接组图谱的延续。目前，我们已经建立了以高通量、工业化的方式制造、收集和处理大规模、多荧光连接数据的整个管道。为了跟踪和组织这些大规模数据，我们开发了实验室信息管理系统（LIMS），加快了数据管理并减少了人为错误。遵循一个公正的，系统的，立体定位的“基于网格”的方法，使用双重共同注射的成熟的神经回路跟踪技术，我们一直在构建一个“骨架”连接组地图（第一阶段），这将构成约1600传入和传出通路从400只动物。在这里提出的第二阶段，我们将使用1200只动物，按照改进的“基于网格”的注射策略，连续生成大规模、超高分辨率的连接性数据。这些注射的收集将产生总共4800个传入通路和4800个传出通路，使整个大脑覆盖。此外，在第一阶段，我们开发了革命性的iConnectome可视化程序（www.MouseConnectome.org），其特征在于可搜索的多荧光神经示踪剂注射目录，可在其自身的亮场Nissl染色的细胞结构背景内以高分辨率查看。iConnectome附带一个全面的连接在线数据库（BAMS），使用户能够以矩阵格式映射复杂的神经元网络，并将其与大鼠，猴子和人类的连接数据相关联。此外，由我们的信息学团队开发的iConnectome注释报告系统（iCARS）将为每个注射部位提供逐层注释信息。强大的信息学工具最终将使用户能够在标准3D解剖框架内比较任何注射部位的连接模式。为了做到这一点，在第二阶段，我们将以图形方式重建连接所有大脑结构的神经通路，并将提供四个版本的小鼠连接组图谱（MCA）：（1）MCA 1.0（谷歌卫星和谷歌街景），用于显示高分辨率多荧光标记的“原始”成像数据;（2）MCA2.0（Google Map）将提供图形化重建的轴突通路和逆行标记的神经元;（3）MCA 3.0（Google driving direction Roadmap）将是连接大脑结构的示意性连接路线图;（4）BAMS生成的神经网络的连接组矩阵和表格式图形表示，也将为MCA 3.0提供文本版导航指令。所有四个版本将同步并集成到同一个框架中，这将使我们能够组装类似Google Earth的最终版本Connectome atlas。虽然是一项艰巨的任务，但我们的第一阶段进展和我们在神经解剖学、脑成像和神经信息学方面拥有世界一流专业知识的团队证明了我们在第三阶段提供完整的三维数字小鼠连接组图谱的能力。

DESCRIPTION (provided by applicant): This proposal is a continuation of our efforts to create a three-dimensional, Google Earth-like, digital Connectome atlas of the C57Black/6J mouse brain. Currently, we have established the entire pipeline for manufacturing, collecting, and processing large-scale, multi-fluorescent connectivity data in a high-throughput, industrialized manner. To track and organize these large-scale data, we have developed the Laboratory Information Management System (LIMS), which expedites data management and reduces human error. Following an unbiased, systematic, stereotaxic "grid-based" approach using double coinjections of proven neural circuit tracing technology, we have been constructing a "skeleton" Connectome map (Phase I), which will constitute about 1600 afferent and 1600 efferent pathways from 400 animals. In Phase II, proposed here, we will continuously generate large-scale, ultra-high-resolution connectivity data following a refined "grid- based" injection strategy using 1200 animals. The collection of these injections will generate a total of 4800 afferent and 4800 efferent pathways, enabling whole brain coverage. Further, in Phase I, we developed the revolutionary iConnectome visualization program (www.MouseConnectome.org) that features a searchable catalog of multi-fluorescent neural tracer injections available to view at high-resolution within their own bright- field Nissl-stained cytoarchitectural background. The iConnectome is accompanied by a comprehensive connectivity online database (BAMS) that enables users to map complex neuronal networks in a matrix format and to correlate them with connectivity data available for the rat, monkey, and human. In addition, the iConnectome Annotation Reporting System (iCARS), developed by our informatics team, will provide level by level annotation information for each injection site. Powerful informatics tools eventually will enable users to compare connectivity patterns of any injection site within a standard 3D anatomic frame. To do this, in Phase II, we will graphically reconstruct neural pathways that link all brain structures and will deliver four versions of the Mouse Connectome Atlas (MCA): (1) MCA1.0 (Google satellite and Google street view) for displaying high- resolution multi-fluorescent labeled "raw" imaging data; (2) MCA2.0 (Google Map) will provide graphically reconstructed axonal pathways and retrogradely labeled neurons; (3) MCA3.0 (Google driving direction Roadmap) will be the schematic connectivity route map that links brain structures; and (4) Connectome matrix and table-like graphic representation of neural networks generated by BAMS, which will also provide the text version navigation instructions for MCA3.0. All four versions will be synchronized and integrated into the same framework, which will enable us to assemble the ultimate version Google Earth-like Connectome atlas. Although an enormous undertaking, our Phase I progress and our team with world-class expertise in neuroanatomy, brain imaging, and neuroinformatics is a testament to our ability to deliver the completed three- dimensional digital mouse Connectome atlas in Phase III.