clozapine and levocabastine

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.

Topics: Adaptor Proteins, Signal Transducing; Animals; Bacterial Proteins; Basolateral Nuclear Complex; Clozapine; Extinction, Psychological; Fear; Gene Expression Regulation; Genes, Reporter; Injections, Intraventricular; Integrases; Intercellular Signaling Peptides and Proteins; Luminescent Proteins; Memory; Mice; Oligopeptides; Optogenetics; Piperidines; Prefrontal Cortex; Receptors, Neurotensin; Sensory Receptor Cells; Sequence Analysis, RNA; Stereotaxic Techniques; Thrombospondins; Thy-1 Antigens; Wnt Proteins