Opto-seq reveals input-specific immediate-early gene induction in ventral tegmental area cell types

Neuron. 2024 Aug 21;112(16):2721-2731.e5. doi: 10.1016/j.neuron.2024.05.026. Epub 2024 Jun 19.

Abstract

The ventral tegmental area (VTA) is a critical node in circuits governing motivated behavior and is home to diverse populations of neurons that release dopamine, gamma-aminobutyric acid (GABA), glutamate, or combinations of these neurotransmitters. The VTA receives inputs from many brain regions, but a comprehensive understanding of input-specific activation of VTA neuronal subpopulations is lacking. To address this, we combined optogenetic stimulation of select VTA inputs with single-nucleus RNA sequencing (snRNA-seq) and highly multiplexed in situ hybridization to identify distinct neuronal clusters and characterize their spatial distribution and activation patterns. Quantification of immediate-early gene (IEG) expression revealed that different inputs activated select VTA subpopulations, which demonstrated cell-type-specific transcriptional programs. Within dopaminergic subpopulations, IEG induction levels correlated with differential expression of ion channel genes. This new transcriptomics-guided circuit analysis reveals the diversity of VTA activation driven by distinct inputs and provides a resource for future analysis of VTA cell types.

Keywords: circuit connectivity; dopamine; immediate-early genes; ion channels; optogenetics; snRNA-seq; transcriptomics; ventral tegmental area.

MeSH terms

  • Animals
  • Dopaminergic Neurons / metabolism
  • Genes, Immediate-Early*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Neurons / metabolism
  • Optogenetics* / methods
  • Sequence Analysis, RNA / methods
  • Ventral Tegmental Area* / cytology
  • Ventral Tegmental Area* / metabolism