title: Functional effects of polymorphisms on glucocorticoid receptor modulation of human anxiogenic substance-P gene promoter activity in primary amygdala neurones.
abstract: Expression or introduction of the neuropeptide substance-P (SP; encoded by the TAC1 gene in humans and Tac1 in rodents) in the amygdala induces anxiety related behaviour in rodents. In addition, pharmacological antagonism of the main receptor of SP in humans; NK1, is anxiolytic. In the current study, we show that the Tac1 locus is up-regulated in primary rat amygdala neurones in response to activation of the glucocorticoid receptor (GR); a classic component of the stress response. Using a combination of bioinformatics, electrophoretic mobility shift assays (EMSA) and reporter plasmid magnetofection into rat primary amygdala neurones we identified a highly conserved GR response sequence (2GR) in the human TAC1 promoter that binds GR in response to dexamethasone (Dex) or forskolin. We also identified a second GR binding site in the human promoter that was polymorphic and whose T-allele is only found in Japanese and Chinese populations. We present evidence that the T-allele of SNPGR increases the activity of the TAC1 promoter through de-sequestration or de-repression of 2GR. The identification of Dex/forskolin response elements in the TAC1 promoter in amygdala neurones suggests a possible link in the chain of molecular events connecting GR activation and anxiety. In addition, the discovery of a SNP which can alter this response may have implications for our understanding of the role of regulatory variation in susceptibility to stress in specific populations.
authors.affiliation: School of Medical Sciences, Institute of Medical Sciences, University of Aberdeen, Aberdeen AB39 3UW, Scotland, UK.
authors.forename: Colin W
authors.initials: CW
authors.lastname: Hay
authors.affiliation: School of Medical Sciences, Institute of Medical Sciences, University of Aberdeen, Aberdeen AB39 3UW, Scotland, UK.
authors.forename: Lynne
authors.initials: L
authors.lastname: Shanley
authors.affiliation: School of Medical Sciences, Institute of Medical Sciences, University of Aberdeen, Aberdeen AB39 3UW, Scotland, UK.
authors.forename: Scott
authors.initials: S
authors.lastname: Davidson
authors.affiliation: School of Medical Sciences, Institute of Medical Sciences, University of Aberdeen, Aberdeen AB39 3UW, Scotland, UK.
authors.forename: Philip
authors.initials: P
authors.lastname: Cowie
authors.affiliation: School of Medical Sciences, Institute of Medical Sciences, University of Aberdeen, Aberdeen AB39 3UW, Scotland, UK.
authors.forename: Marissa
authors.initials: M
authors.lastname: Lear
authors.affiliation: MRC SGDP Centre, Institute of Psychiatry, King's College London, DeCrespigny Park, London SE5 8AF3, UK.
authors.forename: Peter
authors.initials: P
authors.lastname: McGuffin
authors.affiliation: School of Medical Sciences, Institute of Medical Sciences, University of Aberdeen, Aberdeen AB39 3UW, Scotland, UK.
authors.forename: Gernot
authors.initials: G
authors.lastname: Riedel
authors.affiliation: School of Medical Sciences, Institute of Medical Sciences, University of Aberdeen, Aberdeen AB39 3UW, Scotland, UK.
authors.forename: Iain J
authors.initials: IJ
authors.lastname: McEwan
authors.affiliation: School of Medical Sciences, Institute of Medical Sciences, University of Aberdeen, Aberdeen AB39 3UW, Scotland, UK. Electronic address: alasdair.mackenzie@abdn.ac.uk.
authors.forename: Alasdair
authors.initials: A
authors.lastname: MacKenzie
mesh_terms: D000679:Amygdala / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D000818:Animals; D000831:Animals, Newborn; D001007:Anxiety / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D008969:Molecular Sequence Data; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D020641:Polymorphism, Single Nucleotide; D061251:Primary Cell Culture; D011401:Promoter Regions, Genetic / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D011965:Receptors, Glucocorticoid / Q000819:agonists / Q000502:physiology*; D012689:Sequence Homology, Nucleic Acid; D013373:Substance P / Q000235:genetics* / Q000378:metabolism
chemical_list: D011965:Receptors, Glucocorticoid; D005576:Colforsin; D013373:Substance P; D003907:Dexamethasone
keywords: Gene regulation; Glucocorticoid receptor; Single nucleotide polymorphism; Stress and anxiety; TAC1 promoter