title: Overexpression of 5-HT1B receptor in dorsal raphe nucleus using Herpes Simplex Virus gene transfer increases anxiety behavior after inescapable stress.
abstract: 5-HT(1B) autoreceptors have been implicated in animal models of stress and are regulated selectively by serotonin-selective reuptake inhibitors such as fluoxetine. These terminal autoreceptors regulate serotonin release from dorsal raphe nucleus (DRN) projections throughout rat forebrain. However, it has not been previously possible to manipulate 5-HT(1B) autoreceptor activity selectively without also changing 5-HT(1B) activity in other neurons mediating different behavioral responses. Therefore, we have developed a viral-mediated gene transfer strategy to express hemagglutinin-tagged 5-HT(1B) and manipulate these autoreceptors in DRN. Green fluorescent protein (GFP) was coexpressed from a separate transcriptional unit on the same amplicon to assist in monitoring infection and expression. We confirmed the expression and biological activity of both transgenic proteins in vitro. When injected directly into DRN using stereotaxic procedure, HA-5-HT(1B) receptors were expressed in serotonergic neurons and translocated to the forebrain. The effect of DRN expression of HA-5-HT(1B) on stress-induced behaviors was compared with control rats that received GFP-only amplicons. There was no change in immobility in the forced swim test. However, HA-5-HT(1B) expression significantly reduced entrances into the central region of an open-field arena after water-restraint stress without altering overall locomotor activity, but not in the absence of stress exposure. HA-5-HT(1B) expression also reduced entries into the open arms of the elevated plus maze after water restraint. Because these tests are sensitive to increases in anxiety-like behavior, our results suggest that overactivity of 5-HT(1B) autoreceptors in DRN neurons may be an important mediator of pathological responses to stressful events.
authors.affiliation: Department of Psychiatry and Behavioral Sciences and Harborview Medical Center, University of Washington, Seattle, Washington 98195, USA.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Clark
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Sexton
authors.forename: Molly
authors.initials: M
authors.lastname: McClain
authors.forename: Daniel
authors.initials: D
authors.lastname: Root
authors.forename: Ruth
authors.initials: R
authors.lastname: Kohen
authors.forename: John F
authors.initials: JF
authors.lastname: Neumaier
mesh_terms: D000818:Animals; D001007:Anxiety / Q000235:genetics / Q000503:physiopathology*; D001522:Behavior, Animal / Q000187:drug effects / Q000502:physiology; D019556:COS Cells; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism; D004333:Drug Administration Routes; D004742:Enhancer Elements, Genetic; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D049452:Green Fluorescent Proteins; D006388:Hemagglutinins / Q000235:genetics; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D011903:Raphe Nuclei / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D044302:Receptor, Serotonin, 5-HT1B; D011985:Receptors, Serotonin / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D018139:Simplexvirus / Q000235:genetics*; D013037:Spatial Behavior / Q000187:drug effects / Q000502:physiology; D013312:Stress, Physiological / Q000503:physiopathology*; D014162:Transfection
chemical_list: C470302:HTR1B protein, human; D006388:Hemagglutinins; D008164:Luminescent Proteins; D044302:Receptor, Serotonin, 5-HT1B; D011985:Receptors, Serotonin; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D000242:Cyclic AMP