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

描述(由申请人提供)：这项提案是我们为C57Black/6J小鼠大脑创建一个三维的、类似Google Earth的数字连接图集的继续努力。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流水线。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)，该系统加快了数据管理并减少了人为错误。按照一种无偏见的、系统的、立体定位的、基于网格的方法，使用经过验证的神经回路追踪技术的双重联合注射，我们一直在构建一个连接组图(第一阶段)，它将包括来自400个动物的大约1600条传入和1600条传出通路。在这里提出的第二阶段，我们将继续生成大规模、超高分辨率的连通性数据，遵循使用1200只动物的改进的“基于网格的”注入策略。这些注射的集合将产生总计4800条传入和4800条传出通路，使之能够覆盖整个大脑。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)，该程序具有可搜索的多荧光神经示踪剂注射目录，可在其自身明亮的Nissl染色的细胞结构背景下以高分辨率查看。IConnectome配有一个全面的连接性在线数据库(BAMS)，使用户能够以矩阵格式绘制复杂的神经元网络图，并将它们与大鼠、猴子和人类可用的连接性数据相关联。此外，由我们的信息学团队开发的iConnectome注释报告系统(ICars)将提供每个注射部位的逐级注释信息。强大的信息学工具最终将使用户能够比较标准3D解剖框架内任何注射部位的连接模式。为此，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路径，并将提供四个版本的老鼠连接图谱(MCA)：(1)MCA1.0(谷歌卫星和谷歌街景)用于显示高分辨率的多荧光标记的“原始”成像数据；(2)MCA2.0(谷歌地图)将提供图形重建的轴突路径和逆行标记的神经元；(3)MCA3.0(谷歌驾驶方向路线图)将是连接大脑结构的示意图连接路线图；(4)BAMS生成的神经网络的连接矩阵和表状图形表示，它还将为MCA3.0提供文本版本的导航指令。所有四个版本将被同步并集成到同一个框架中，这将使我们能够组装出最终版本，类似谷歌地球的Connectome地图集。尽管这是一项巨大的事业，但我们的第一阶段取得了进展，我们的团队在神经解剖学、脑成像和神经信息学方面拥有世界级的专业知识，这证明了我们有能力在第三阶段提供完整的三维数字鼠标Connectome图谱。

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.