Voltage-Seq: All-optical Postsynaptic Connectome-guided Single-cell Transcriptomics

Overview

Journal Nat Methods

Specialties Biomedical Engineering
Pathology

Date 2023 Jul 20

PMID 37474808

Authors

Veronika Csillag

Marianne Hiriart Bizzozzero

J C Noble

Bjorn Reinius

Janos Fuzik

Affiliations

Soon will be listed here.

Abstract

Understanding the routing of neuronal information requires the functional characterization of connections. Neuronal projections recruit large postsynaptic ensembles with distinct postsynaptic response types (PRTs). PRT is typically probed by low-throughput whole-cell electrophysiology and is not a selection criterion for single-cell RNA-sequencing (scRNA-seq). To overcome these limitations and target neurons based on specific PRTs for soma harvesting and subsequent scRNA-seq, we created Voltage-Seq. We established all-optical voltage imaging and recorded the PRT of 8,347 neurons in the mouse periaqueductal gray (PAG) evoked by the optogenetic activation of ventromedial hypothalamic (VMH) terminals. PRTs were classified and spatially resolved in the entire VMH-PAG connectome. We built an onsite analysis tool named VoltView to navigate soma harvesting towards target PRTs guided by a classifier that used the VMH-PAG connectome database as a reference. We demonstrated Voltage-seq by locating VMH-driven γ-aminobutyric acid-ergic neurons in the PAG, guided solely by the onsite classification in VoltView.

Citing Articles

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

Csillag V, Noble J, Calvigioni D, Reinius B, Fuzik J Nat Protoc. 2024; 19(10):2863-2890.

PMID: 38834919 DOI: 10.1038/s41596-024-01005-y.

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