Aug 23, 2016

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

Neuron
Justus M KebschullAnthony M Zador

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.

  • References35
  • Citations54

Mentioned in this Paper

Projections and Predictions
Sequence Determinations, RNA
Neurons
Brain
Neuroanatomy
Research Project
Nucleic Acid Sequencing
Structure of Locus Ceruleus
Structure of Cortex of Kidney
Sequencing

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