Molecular characterization of Thy1 expressing fear-inhibiting neurons within the basolateral amygdala

Nat Commun. 2016 Oct 21:7:13149. doi: 10.1038/ncomms13149.

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

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adaptor Proteins, Signal Transducing
  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Basolateral Nuclear Complex / cytology
  • Basolateral Nuclear Complex / drug effects
  • Basolateral Nuclear Complex / metabolism*
  • Clozapine / analogs & derivatives
  • Clozapine / pharmacology
  • Extinction, Psychological / drug effects
  • Extinction, Psychological / physiology*
  • Fear / drug effects
  • Fear / physiology*
  • Gene Expression Regulation
  • Genes, Reporter
  • Injections, Intraventricular
  • Integrases / genetics
  • Integrases / metabolism
  • Intercellular Signaling Peptides and Proteins / genetics
  • Intercellular Signaling Peptides and Proteins / metabolism
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Memory / drug effects
  • Memory / physiology*
  • Mice
  • Oligopeptides / pharmacology
  • Optogenetics / methods
  • Piperidines / pharmacology
  • Prefrontal Cortex / cytology
  • Prefrontal Cortex / drug effects
  • Prefrontal Cortex / metabolism
  • Receptors, Neurotensin / genetics
  • Receptors, Neurotensin / metabolism
  • Sensory Receptor Cells / cytology
  • Sensory Receptor Cells / drug effects
  • Sensory Receptor Cells / metabolism
  • Sequence Analysis, RNA
  • Stereotaxic Techniques
  • Thrombospondins / genetics
  • Thrombospondins / metabolism
  • Thy-1 Antigens / genetics*
  • Thy-1 Antigens / metabolism
  • Wnt Proteins / genetics
  • Wnt Proteins / metabolism

Substances

  • Adaptor Proteins, Signal Transducing
  • Bacterial Proteins
  • Dkk3 protein, mouse
  • Intercellular Signaling Peptides and Proteins
  • Luminescent Proteins
  • Ntsr2 protein, mouse
  • Oligopeptides
  • Piperidines
  • RSPO2 protein, mouse
  • Receptors, Neurotensin
  • Thrombospondins
  • Thy-1 Antigens
  • Wnt Proteins
  • Wnt7a protein, mouse
  • beta-lactotensin
  • yellow fluorescent protein, Bacteria
  • Cre recombinase
  • Integrases
  • levocabastine
  • Clozapine
  • clozapine N-oxide