title: Visualisation and characterisation of oestrogen receptor α-positive neurons expressing green fluorescent protein under the control of the oestrogen receptor α promoter.
abstract: Oestrogen receptor (ER)α plays important roles in the development and function of various neuronal systems through activation by its ligands, oestrogens. To visualise ERα-positive neurons, we generated transgenic (tg) mice expressing green fluorescent protein (GFP) under the control of the ERα promoter. In three independent tg lines, GFP-positive neurons were observed in areas previously reported to express ERα mRNA, including the lateral septum, bed nucleus of the stria terminalis, medial preoptic nucleus (MPO), hypothalamus, and amygdala. In these areas, GFP signals mostly overlapped with ERα immunoreactivity. GFP fluorescence was seen in neurites and cell bodies of neurons. In addition, the network and detailed structure of neurites were visible in dissociated and slice cultures of hypothalamic neurons. We examined the effect of oestrogen deprivation by ovariectomy on the structure of the GFP-positive neurons. The area of ERα-positive cell bodies in the bed nucleus of the stria terminalis and MPO was measured by capturing the GFP signal and was found to be significantly smaller in ovariectomy mice than in control mice. When neurons in the MPO were infected with an adeno-associated virus that expressed small hairpin RNA targeting the ERα gene, an apparent induction of GFP was observed in this area, suggesting a negative feedback mechanism in which ERα controls expression of the ERα gene itself. Thus, the ERα promoter-GFP tg mice will be useful to analyse the development and plastic changes of the structure of ERα-expressing neurons and oestrogen and its receptor-mediated neuronal responses.
authors.affiliation: Department of Anatomy and Neurobiology, Kyoto Prefectural University of Medicine Graduate School of Medical Science, 465 Kawaramachi Hirokoji, Kamigyoku, Kyoto 602-8566, Japan. matsuken@koto.kpu-m.ac.jp
authors.forename: Ken Ichi
authors.initials: KI
authors.lastname: Matsuda
authors.forename: Miho
authors.initials: M
authors.lastname: Yanagisawa
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Sano
authors.forename: Ikuo
authors.initials: I
authors.lastname: Ochiai
authors.forename: Sergei
authors.initials: S
authors.lastname: Musatov
authors.forename: Kota
authors.initials: K
authors.lastname: Okoshi
authors.forename: Shinji
authors.initials: S
authors.lastname: Tsukahara
authors.forename: Sonoko
authors.initials: S
authors.lastname: Ogawa
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Kawata
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology / Q000378:metabolism*; D002478:Cells, Cultured; D047628:Estrogen Receptor alpha / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D005260:Female; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009474:Neurons / Q000378:metabolism*; D010052:Ovariectomy; D011401:Promoter Regions, Genetic
chemical_list: D047628:Estrogen Receptor alpha; D049452:Green Fluorescent Proteins
keywords: adeno-associated virus; green fluorescent protein; ovarian steroid hormone; small hairpin RNA; transgenic mouse