title: Acute engagement of Gq-mediated signaling in the bed nucleus of the stria terminalis induces anxiety-like behavior.
abstract: The bed nucleus of the stria terminalis (BNST) is a brain region important for regulating anxiety-related behavior in both humans and rodents. Here we used a chemogenetic strategy to investigate how engagement of G protein-coupled receptor (GPCR) signaling cascades in genetically defined GABAergic BNST neurons modulates anxiety-related behavior and downstream circuit function. We saw that stimulation of vesicular γ-aminobutyric acid (GABA) transporter (VGAT)-expressing BNST neurons using hM3Dq, but neither hM4Di nor rM3Ds designer receptors exclusively activated by a designer drug (DREADD), promotes anxiety-like behavior. Further, we identified that activation of hM3Dq receptors in BNST VGAT neurons can induce a long-term depression-like state of glutamatergic synaptic transmission, indicating DREADD-induced changes in synaptic plasticity. Further, we used DREADD-assisted metabolic mapping to profile brain-wide network activity following activation of Gq-mediated signaling in BNST VGAT neurons and saw increased activity within ventral midbrain structures, including the ventral tegmental area and hindbrain structures such as the locus coeruleus and parabrachial nucleus. These results highlight that Gq-mediated signaling in BNST VGAT neurons can drive downstream network activity that correlates with anxiety-like behavior and points to the importance of identifying endogenous GPCRs within genetically defined cell populations. We next used a microfluidics approach to profile the receptorome of single BNST VGAT neurons. This approach yielded multiple Gq-coupled receptors that are associated with anxiety-like behavior and several potential novel candidates for regulation of anxiety-like behavior. From this, we identified that stimulation of the Gq-coupled receptor 5-HT2CR in the BNST is sufficient to elevate anxiety-like behavior in an acoustic startle task. Together, these results provide a novel profile of receptors within genetically defined BNST VGAT neurons that may serve as therapeutic targets for regulating anxiety states and provide a blueprint for examining how G-protein-mediated signaling in a genetically defined cell type can be used to assess behavior and brain-wide circuit function.
authors.affiliation: Neurobiology Curriculum, University of North Carolina, Chapel Hill, NC, USA.|Laboratory of Molecular Neurophysiology, Bowles Center for Alcohol Studies, University of North Carolina School of Medicine, Chapel Hill, NC, USA.
authors.forename: C M
authors.initials: CM
authors.lastname: Mazzone
authors.affiliation: Laboratory of Molecular Neurophysiology, Bowles Center for Alcohol Studies, University of North Carolina School of Medicine, Chapel Hill, NC, USA.
authors.forename: D
authors.initials: D
authors.lastname: Pati
authors.affiliation: Fishberg Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, NY, USA.|Department of Psychiatry, Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA.|Biobehavioral Imaging and Molecular Neuropsychopharmacology Unit, Neuroimaging Research Branch, National Institute on Drug Abuse, Baltimore, MD, USA.
authors.forename: M
authors.initials: M
authors.lastname: Michaelides
authors.affiliation: Laboratory of Molecular Neurophysiology, Bowles Center for Alcohol Studies, University of North Carolina School of Medicine, Chapel Hill, NC, USA.
authors.forename: J
authors.initials: J
authors.lastname: DiBerto
authors.affiliation: Department of Psychology, University of Vermont, Burlington, VT, USA.
authors.forename: J H
authors.initials: JH
authors.lastname: Fox
authors.affiliation: Laboratory of Molecular Neurophysiology, Bowles Center for Alcohol Studies, University of North Carolina School of Medicine, Chapel Hill, NC, USA.
authors.forename: G
authors.initials: G
authors.lastname: Tipton
authors.affiliation: Center for Gastrointestinal Biology and Disease, University of North Carolina, Chapel Hill, NC, USA.
authors.forename: C
authors.initials: C
authors.lastname: Anderson
authors.affiliation: Laboratory of Molecular Neurophysiology, Bowles Center for Alcohol Studies, University of North Carolina School of Medicine, Chapel Hill, NC, USA.
authors.forename: K
authors.initials: K
authors.lastname: Duffy
authors.affiliation: Laboratory of Molecular Neurophysiology, Bowles Center for Alcohol Studies, University of North Carolina School of Medicine, Chapel Hill, NC, USA.
authors.forename: J M
authors.initials: JM
authors.lastname: McKlveen
authors.affiliation: Laboratory of Molecular Neurophysiology, Bowles Center for Alcohol Studies, University of North Carolina School of Medicine, Chapel Hill, NC, USA.
authors.forename: J A
authors.initials: JA
authors.lastname: Hardaway
authors.affiliation: Department of Medicine, University of North Carolina, Chapel Hill, NC, USA.|Department of Cell Biology and Physiology, and Biomedical Engineering, University of North Carolina, Chapel Hill, NC, USA.
authors.forename: S T
authors.initials: ST
authors.lastname: Magness
authors.affiliation: Department of Psychology, University of Vermont, Burlington, VT, USA.
authors.forename: W A
authors.initials: WA
authors.lastname: Falls
authors.affiliation: Department of Psychology, University of Vermont, Burlington, VT, USA.
authors.forename: S E
authors.initials: SE
authors.lastname: Hammack
authors.affiliation: Department of Psychiatry, University of North Carolina, Chapel Hill, NC, USA.
authors.forename: Z A
authors.initials: ZA
authors.lastname: McElligott
authors.affiliation: Fishberg Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, NY, USA.|Department of Psychiatry, Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA.
authors.forename: Y L
authors.initials: YL
authors.lastname: Hurd
authors.affiliation: Laboratory of Molecular Neurophysiology, Bowles Center for Alcohol Studies, University of North Carolina School of Medicine, Chapel Hill, NC, USA.|Department of Pharmacology, School of Medicine, University of North Carolina at Chapel Hill, NC, USA.
authors.forename: T L
authors.initials: TL
authors.lastname: Kash
mesh_terms: D000818:Animals; D014151:Anti-Anxiety Agents / Q000627:therapeutic use; D001007:Anxiety / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology*; D001931:Brain Mapping; D063387:Cannabinoid Receptor Antagonists / Q000494:pharmacology; D003024:Clozapine / Q000031:analogs & derivatives / Q000494:pharmacology; D003623:Dark Adaptation / Q000187:drug effects / Q000235:genetics; D004195:Disease Models, Animal; D004963:Estrenes / Q000494:pharmacology; D019706:Excitatory Postsynaptic Potentials / Q000187:drug effects / Q000235:genetics; D005106:Exploratory Behavior / Q000187:drug effects; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D066298:In Vitro Techniques; D008297:Male; D018782:Maze Learning / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D010879:Piperazines / Q000494:pharmacology; D011760:Pyrrolidinones / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D011955:Receptors, Drug / Q000187:drug effects / Q000502:physiology; D000077285:Rimonabant / Q000494:pharmacology; D012686:Septal Nuclei / Q000378:metabolism / Q000473:pathology*; D017366:Serotonin Receptor Agonists / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D026941:Sodium Channel Blockers / Q000494:pharmacology; D013779:Tetrodotoxin / Q000627:therapeutic use; D050495:Vesicular Inhibitory Amino Acid Transport Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D014151:Anti-Anxiety Agents; D063387:Cannabinoid Receptor Antagonists; D004963:Estrenes; D010726:Phosphodiesterase Inhibitors; D010879:Piperazines; D011760:Pyrrolidinones; D012333:RNA, Messenger; D011955:Receptors, Drug; D017366:Serotonin Receptor Agonists; D026941:Sodium Channel Blockers; D050495:Vesicular Inhibitory Amino Acid Transport Proteins; C492244:Viaat protein, mouse; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; D049452:Green Fluorescent Proteins; D013779:Tetrodotoxin; D043802:GTP-Binding Protein alpha Subunits, Gq-G11; D003024:Clozapine; C079149:clozapine N-oxide; C015068:1-(3-chlorophenyl)piperazine; D000077285:Rimonabant