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Neuron. 2016 Sep 7;91(5):975-987. doi: 10.1016/j.neuron.2016.07.036. Epub 2016 Aug 18.

High-Throughput Mapping of Single-Neuron Projections by Sequencing of Barcoded RNA.

Author information

1
Watson School of Biological Sciences, Cold Spring Harbor, NY 11724, USA; Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
2
Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA; Champalimaud Centre for the Unknown, 1400-038 Lisbon, Portugal.
3
Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
4
Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA. Electronic address: zador@cshl.edu.

Abstract

Neurons transmit information to distant brain regions via long-range axonal projections. In the mouse, area-to-area connections have only been systematically mapped using bulk labeling techniques, which obscure the diverse projections of intermingled single neurons. Here we describe MAPseq (Multiplexed Analysis of Projections by Sequencing), a technique that can map the projections of thousands or even millions of single neurons by labeling large sets of neurons with random RNA sequences ("barcodes"). Axons are filled with barcode mRNA, each putative projection area is dissected, and the barcode mRNA is extracted and sequenced. Applying MAPseq to the locus coeruleus (LC), we find that individual LC neurons have preferred cortical targets. By recasting neuroanatomy, which is traditionally viewed as a problem of microscopy, as a problem of sequencing, MAPseq harnesses advances in sequencing technology to permit high-throughput interrogation of brain circuits.

PMID:
27545715
DOI:
10.1016/j.neuron.2016.07.036
[Indexed for MEDLINE]
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