Abstract
Molecular characterization of neuron populations, particularly those controlling threat responses, is essential for understanding the cellular basis of behaviour and identifying pharmacological agents acting selectively on fear-controlling circuitry. Here we demonstrate a comprehensive workflow for identification of pharmacologically tractable markers of behaviourally characterized cell populations. Thy1-eNpHR-, Thy1-Cre- and Thy1-eYFP-labelled neurons of the BLA consistently act as fear inhibiting or 'Fear-Off' neurons during behaviour. We use cell-type-specific optogenetics and chemogenetics (DREADDs) to modulate activity in this population during behaviour to block or enhance fear extinction. Dissociated Thy1-eYFP neurons are isolated using FACS. RNA sequencing identifies genes strongly upregulated in RNA of this population, including Ntsr2, Dkk3, Rspo2 and Wnt7a. Pharmacological manipulation of neurotensin receptor 2 confirms behavioural effects observed in optogenetic and chemogenetic experiments. These experiments identify and validate Ntsr2-expressing neurons within the BLA, as a putative 'Fear-Off' population.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Adaptor Proteins, Signal Transducing
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Animals
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Bacterial Proteins / genetics
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Bacterial Proteins / metabolism
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Basolateral Nuclear Complex / cytology
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Basolateral Nuclear Complex / drug effects
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Basolateral Nuclear Complex / metabolism*
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Clozapine / analogs & derivatives
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Clozapine / pharmacology
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Extinction, Psychological / drug effects
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Extinction, Psychological / physiology*
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Fear / drug effects
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Fear / physiology*
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Gene Expression Regulation
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Genes, Reporter
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Injections, Intraventricular
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Integrases / genetics
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Integrases / metabolism
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Intercellular Signaling Peptides and Proteins / genetics
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Intercellular Signaling Peptides and Proteins / metabolism
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Luminescent Proteins / genetics
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Luminescent Proteins / metabolism
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Memory / drug effects
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Memory / physiology*
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Mice
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Oligopeptides / pharmacology
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Optogenetics / methods
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Piperidines / pharmacology
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Prefrontal Cortex / cytology
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Prefrontal Cortex / drug effects
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Prefrontal Cortex / metabolism
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Receptors, Neurotensin / genetics
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Receptors, Neurotensin / metabolism
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Sensory Receptor Cells / cytology
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Sensory Receptor Cells / drug effects
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Sensory Receptor Cells / metabolism
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Sequence Analysis, RNA
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Stereotaxic Techniques
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Thrombospondins / genetics
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Thrombospondins / metabolism
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Thy-1 Antigens / genetics*
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Thy-1 Antigens / metabolism
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Wnt Proteins / genetics
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Wnt Proteins / metabolism
Substances
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Adaptor Proteins, Signal Transducing
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Bacterial Proteins
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Dkk3 protein, mouse
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Intercellular Signaling Peptides and Proteins
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Luminescent Proteins
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Ntsr2 protein, mouse
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Oligopeptides
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Piperidines
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RSPO2 protein, mouse
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Receptors, Neurotensin
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Thrombospondins
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Thy-1 Antigens
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Wnt Proteins
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Wnt7a protein, mouse
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beta-lactotensin
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yellow fluorescent protein, Bacteria
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Cre recombinase
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Integrases
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levocabastine
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Clozapine
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clozapine N-oxide