High-Throughput Mapping of Long-Range Neuronal Projection Using In Situ Sequencing

Xiaoyin Chen, Yu Chi Sun, Huiqing Zhan, Justus M. Kebschull, Stephan Fischer, Katherine Matho, Z. Josh Huang, Jesse Gillis, Anthony M. Zador

Research output: Contribution to journalArticlepeer-review

21 Scopus citations


Understanding neural circuits requires deciphering interactions among myriad cell types defined by spatial organization, connectivity, gene expression, and other properties. Resolving these cell types requires both single-neuron resolution and high throughput, a challenging combination with conventional methods. Here, we introduce barcoded anatomy resolved by sequencing (BARseq), a multiplexed method based on RNA barcoding for mapping projections of thousands of spatially resolved neurons in a single brain and relating those projections to other properties such as gene or Cre expression. Mapping the projections to 11 areas of 3,579 neurons in mouse auditory cortex using BARseq confirmed the laminar organization of the three top classes (intratelencephalic [IT], pyramidal tract-like [PT-like], and corticothalamic [CT]) of projection neurons. In depth analysis uncovered a projection type restricted almost exclusively to transcriptionally defined subtypes of IT neurons. By bridging anatomical and transcriptomic approaches at cellular resolution with high throughput, BARseq can potentially uncover the organizing principles underlying the structure and formation of neural circuits.

Original languageEnglish (US)
Pages (from-to)772-786.e19
Issue number3
StatePublished - Oct 17 2019
Externally publishedYes


  • auditory cortex
  • cellular barcoding
  • high throughput
  • in situ sequencing
  • projection mapping

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)


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