Transcriptional profiling aligned with in situ expression image analysis reveals mosaically expressed molecular markers for GABA neuron sub-groups in the ventral tegmental area.

The European Journal of Neuroscience
Eleanor J PaulMark A Ungless

Abstract

γ-Aminobutyric acid (GABA) neurons in the ventral tegmental area (VTA) provide local inhibitory control of dopamine neuron activity and send long-range projections to several target regions including the nucleus accumbens. They play diverse roles in reward and aversion, suggesting that they be comprised of several functionally distinct sub-groups, but our understanding of this diversity has been limited by a lack of molecular markers that might provide genetic entry points for cell type-specific investigations. To address this, we conducted transcriptional profiling of GABA neurons and dopamine neurons using immunoprecipitation of tagged polyribosomes (RiboTag) and RNAseq. First, we directly compared these two transcriptomes in order to obtain a list of genes enriched in GABA neurons compared with dopamine neurons. Next, we created a novel bioinformatic approach, that used the PANTHER (Protein ANalysis THrough Evolutionary Relationships) gene ontology database and VTA gene expression data from the Allen Mouse Brain Atlas, from which we obtained 6 candidate genes: Cbln4, Rxfp3, Rora, Gpr101, Trh and Nrp2. As a final step, we verified the selective expression of these candidate genes in sub-groups of GABA neurons in the VTA (and ...Continue Reading

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Citations

Dec 24, 2019·Frontiers in Neural Circuits·Chloé BouarabAbigail M Polter
Nov 23, 2020·Journal of Neurochemistry·Alan FlanaganJonathan D Johnston
May 17, 2021·Neuropharmacology·Patrick C Andrews, Shashank M Dravid

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Methods Mentioned

BETA
immunoprecipitation
RNAseq
electrophoresis
chip
immunoprecipitations

Key Resources (RRID) Mentioned

IMSR_JAX

Software Mentioned

Fiji
DESeq2 Bioconductor
HTSeq
Tophat splice junction mapper
PANTHER
Allen
DESeq2
Ensembl
RiboTag
ImageJ

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