Journal Article

Massive multiplexing of spatially resolved single neuron projections with axonal BARseq

Yuan, Li, Zhan, Huiqing, Chen, Xiaoyin, Henry, Gilbert L., Zador, Anthony M.

2024

Overview

Neurons in the cortex are heterogeneous, sending diverse axonal projections to multiple brain regions. Unraveling the logic of these projections requires single-neuron resolution. Although a growing number of techniques have enabled high-throughput reconstruction, these techniques are typically limited to dozens or at most hundreds of neurons per brain, requiring that statistical analyses combine data from different specimens. Here we present axonal BARseq, a high-throughput approach based on reading out nucleic acid barcodes using in situ RNA sequencing, which enables analysis of even densely labeled neurons. As a proof of principle, we have mapped the long-range projections of >8000 primary auditory cortex neurons from a single male mouse. We identified major cell types based on projection targets and axonal trajectory. The large sample size enabled us to systematically quantify the projections of intratelencephalic (IT) neurons, and revealed that individual IT neurons project to different layers in an area-dependent fashion. Axonal BARseq is a powerful technique for studying the heterogeneity of single neuronal projections at high throughput within individual brains. High-throughput mapping of neural projections at the single-cell level remains challenging. Here, the authors have developed axonal BARseq, a new technique that allows for mapping projections from thousands of neurons within a single brain.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 38/23 / 631/378/2619/2618 / 631/378/3920 / Animals / Auditory Cortex - cytology / Auditory Cortex - physiology / Axon guidance / Axons - metabolism / Brain / Brain mapping / Cell size / Cortex (auditory) / Gene sequencing / Heterogeneity / High-Throughput Nucleotide Sequencing - methods / Humanities and Social Sciences / Male / Mapping / Mice / Mice, Inbred C57BL / multidisciplinary / Multiplexing / Neurons / Neurons - cytology / Nucleic acids / Science / Science (multidisciplinary) / Sequence Analysis, RNA - methods / Single-Cell Analysis - methods / Statistical analysis