Kavli Affiliate: Justus M Kebschull
| Authors: Hyopil Kim, Cheng Xu, Craig Washington, Maggie Lowman and Justus M Kebschull
| Summary:
Brain-wide neural circuits are formed by the diverse axonal branching patterns of many individual neurons. Here we introduce POlNTseq (projections of interest by sequencing), a high-throughput and user-friendly barcoded connectomics method that uses cell type specific barcoding and sequencing to rapidly map single-cell projections of a cell type of interest for thousands of neurons per animal. POlNTseq leverages pseudotyping of Sindbis virus and a specific alphavirus- cellular receptor pair to make Sindbis infections cell type specific. lt thus integrates MAPseq-style high-throughput barcoded projection mapping with the established viral-genetic neural circuit analysis toolbox. We validated POlNTseq by mapping genetically and projection-defined cell populations in the mouse motor cortex. We then applied POlNTseq to midbrain dopaminergic neurons and reconstructed the brain-wide single-cell projections of 3,813 dopaminergic neurons in ventral tegmental area (VTA) and substantia nigra pars compacta (SNc). We define over 30 connectomic cell types, vastly exceeding the known diversity of dopaminergic cell types, and identify stereotyped projection motifs that may mediate parallel dopamine signaling. This data constitutes the anatomical substrate on which the diverse functions of dopamine in the brain are built.