BRAIN CONNECTS: Comprehensive regional projection map of marmoset with single axon and cell type resolution

摘要 这一雄心勃勃的提议将建立一个集成的实验-计算平台，以创建第一个 非人灵长类动物--常见的绒猴--的全脑中尺度连接图 (Callithrix Jacchus)、它将通过追踪全脑RNA条形码识别的神经元的轴突投射来做到这一点 绒猴，利用一种基于测序的成像方法，也允许同时转录细胞 对已鉴定的神经元进行分型。这将有助于弥合全脑中尺度连通性数据之间的差距 可供鼠标使用现代技术绘制地图的努力已有十年之久 在人类和NHP中的可比数据。 该提案将把基于病毒条形码的新方法与现有的基于示踪剂注射的方法结合起来 收集史无前例的数据集的方法，这将允许全面绘制区域到区域的地图 绒猴大脑中具有细胞类型特异性的轴突投射，将这些数据应用于以下科学问题 灵长类动物和啮齿动物之间的大脑连接有何不同，解决了MRI衍生的 采取措施将事实联系起来，并利用基于网络的工具让研究界参与到 对生成的数据进行注释和使用。由世界领先的专家和该问题的先驱组成的多元化团队 已经在协作的领域，跨越不同的机构和学科，并配备 优秀的设施和资源，将汇聚在一起，汇聚成赋能平台。有五个具体的 旨在实现既定目标。目标一是建立中尺度投影测图的数据采集平台 在绒猴中使用条形码+荧光AAV/AAV-Retro。目标2是开发全脑范围的单一轴突投影 使用全身注射和同步转录切割细胞分型的图谱。目标3是开发数据 管理和分析管道。目标4是进行中尺度连通性的比较研究 绒猴和小鼠，将基于MRI的连接与地面事实进行比较。目标5是为 数据传播、通过门户网站的社区参与、协作校对和注释。 这样的数据集将具有深远的科学意义，可以与第一张灵长类动物的全基因组图相媲美。 正如基因组编码了解有机体表型的蓝图一样，大脑的结构也是如此 电路包含解释动物行为的蓝图，并掌握着许多悬而未决的问题的关键。

SUMMARY This ambitious proposal will establish an integrated experimental-computational platform to create the first comprehensive brain-wide mesoscale connectivity map in a non-human primate, the common marmoset (Callithrix jacchus),. It will do so by tracing axonal projections of RNA barcode-identified neurons brain-wide in the marmoset, utilizing a sequencing-based imaging method that also permits simultaneous transcriptomic cell typing of the identified neurons. This will help bridge the gap between brain-wide mesoscale connectivity data available for the mouse from a decade of mapping efforts using modern techniques and the absence of comparable data in humans and NHP. The proposal will bring together new viral barcode-based approaches with established tracer-injection based methods to collect an unprecedented data set which will permit comprehensive mapping of region-to-region axonal projections in the marmoset brain with cell-type specificity, apply this data to scientific questions regarding how brain connectivity differs between primates and rodents, address the relation between MRI-derived measures to ground-truth connectivity, and utilize web-based tools to engage the research community in annotating and using the resulting data. A diverse team of world-leading experts and pioneers in the problem domain who are already collaborating, spanning diverse institutions and disciplines, and equipped with outstanding facilities and resources, will come together to assemble the enabling platform. There are five specific aims to achieve stated goals. Aim 1 is to establish data acquisition platform for mesoscale projection mapping in marmoset using barcoded + fluorescent AAV/AAV-retro. Aim 2 is to develop brain-wide single-axon projection maps using systemic injections with simultaneous transcriptomic cell-typing. Aim 3 is to develop data management and analysis pipelines. Aim 4 is to perform comparative studies of the mesoscale connectivity in marmoset and mouse, comparing MRI-based connectivity with ground truth. Aim 5 is to develop platforms for data dissemination, community engagement through web portal, collaborative proofreading and annotations. Such a data set will have profound scientific significance, comparable to the first whole genome map of a primate. As genomes encode the blueprint for understanding an organism’s phenotype, so does the architecture of brain circuitry contain the blueprint for explaining an animal’s behavior and holds the key to many open questions.