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Related Experiment Video

Updated: Jun 24, 2025

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All-optical voltage imaging-guided postsynaptic single-cell transcriptome profiling with Voltage-Seq.

Veronika Csillag1, J C Noble2, Daniela Calvigioni1

  • 1Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden.

Nature Protocols
|June 4, 2024
PubMed
Summary
This summary is machine-generated.

Researchers developed Voltage-Seq, a high-throughput method for analyzing neuronal connections. This technique enables single-cell RNA sequencing of specific postsynaptic response types (PRTs) to understand brain circuitry and gene expression.

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Genetics

Background:

  • Neuronal pathways utilize distinct postsynaptic response types (PRTs) to maintain connections.
  • Selecting neurons based on PRTs for single-cell RNA sequencing was previously limited by low-throughput electrophysiology.

Purpose of the Study:

  • To develop a high-throughput method for targeting neurons based on specific PRTs for single-cell RNA sequencing.
  • To enable detailed analysis of neuronal connectivity and gene expression within specific cell types.

Main Methods:

  • Developed Voltage-Seq, utilizing the genetically encoded voltage indicator Voltron in mouse brain slices.
  • Created VoltView, an analysis tool with a classifier for guiding soma collection of specific PRTs.
  • Detailed procedures for optical path preparation, imaging, analysis, and sequencing library preparation.

Main Results:

  • Voltage-Seq enables high-throughput all-optical synaptic assays and single-cell transcriptomic data acquisition from selected postsynaptic neurons.
  • The method allows for the resolution of connectivity ratios and exploration of PRT diversity within connectomes.
  • Facilitates investigation of correlations between connectivity and gene expression in a cell-type-specific manner.

Conclusions:

  • Voltage-Seq offers a powerful, high-throughput approach to study neuronal connectivity and gene expression.
  • This technique overcomes previous limitations, providing new insights into the diversity of postsynaptic response types.
  • Enables cell-type-specific analysis of excitatory and inhibitory connections and their relationship with gene expression.