title: Identification of hypothalamic arcuate nucleus-specific enhancer region of Kiss1 gene in mice.
abstract: Pulsatile secretion of GnRH plays a pivotal role in follicular development via stimulating tonic gonadotropin secretion in mammals. Kisspeptin neurons, located in the arcuate nucleus (ARC), are considered to be an intrinsic source of the GnRH pulse generator. The present study aimed to determine ARC-specific enhancer(s) of the Kiss1 gene by an in vivo reporter assay. Three green fluorescent protein (GFP) reporter constructs (long, medium length, and short) were generated by insertion of GFP cDNA at the Kiss1 locus. Transgenic female mice bearing the long and medium-length constructs showed apparent GFP signals in kisspeptin-immunoreactive cells in both the ARC and anteroventral periventricular nucleus, in which another population of kisspeptin neurons are located. On the other hand, transgenic mice bearing 5'-truncated short construct showed few GFP signals in the ARC kisspeptin-immunoreactive cells, whereas they showed colocalization of GFP- and kisspeptin-immunoreactivities in the anteroventral periventricular nucleus. In addition, chromatin immunoprecipitation and chromosome conformation capture assays revealed recruitment of unoccupied estrogen receptor-α in the 5'-upstream region and intricate chromatin loop formation between the 5'-upstream and promoter regions of Kiss1 locus in the ARC. Taken together, the present results indicate that 5'-upstream region of Kiss1 locus plays a critical role in Kiss1 gene expression in an ARC-specific manner and that the recruitment of estrogen receptor-α and formation of a chromatin loop between the Kiss1 promoter and the 5' enhancer region may be required for the induction of ARC-specific Kiss1 gene expression. These results suggest that the 5'-upstream region of Kiss1 locus functions as an enhancer for ARC Kiss1 gene expression in mice.
authors.affiliation: Graduate School of Bioagricultural Sciences (T.G., J.T., K.I., S.M., H.A., T.F., K.Ta., H.T., Y.U.), Nagoya University, Nagoya 464-8601, Japan; National Institute for Physiological Sciences (T.G., M.S., K.To., M.H.), Okazaki 444-8585, Japan; Graduate School of Medical Sciences (T.I.), Kyushu University, Fukuoka 812-8581, Japan; Kochi Medical School (K.To.), Kochi University, Kochi 783-8505, Japan; and Graduate School of Agricultural and Life Sciences (K.M.), The University of Tokyo, Tokyo 113-8657, Japan.
authors.forename: Teppei
authors.initials: T
authors.lastname: Goto
authors.forename: Junko
authors.initials: J
authors.lastname: Tomikawa
authors.forename: Kana
authors.initials: K
authors.lastname: Ikegami
authors.forename: Shiori
authors.initials: S
authors.lastname: Minabe
authors.forename: Hitomi
authors.initials: H
authors.lastname: Abe
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Fukanuma
authors.forename: Takuya
authors.initials: T
authors.lastname: Imamura
authors.forename: Kenji
authors.initials: K
authors.lastname: Takase
authors.forename: Makoto
authors.initials: M
authors.lastname: Sanbo
authors.forename: Koichi
authors.initials: K
authors.lastname: Tomita
authors.forename: Masumi
authors.initials: M
authors.lastname: Hirabayashi
authors.forename: Kei-ichiro
authors.initials: K
authors.lastname: Maeda
authors.forename: Hiroko
authors.initials: H
authors.lastname: Tsukamura
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Uenoyama
mesh_terms: D000818:Animals; D001111:Arcuate Nucleus of Hypothalamus / Q000378:metabolism*; D001665:Binding Sites / Q000235:genetics; D002843:Chromatin / Q000235:genetics; D047369:Chromatin Immunoprecipitation; D004268:DNA-Binding Proteins / Q000378:metabolism; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D047628:Estrogen Receptor alpha / Q000235:genetics*; D005260:Female; D015870:Gene Expression; D005786:Gene Expression Regulation / Q000235:genetics*; D017930:Genes, Reporter / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D007032:Hypothalamus, Anterior / Q000378:metabolism; D059648:Kisspeptins / Q000096:biosynthesis / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic / Q000235:genetics; D011487:Protein Conformation
chemical_list: D002843:Chromatin; D004268:DNA-Binding Proteins; D047628:Estrogen Receptor alpha; C491407:Kiss1 protein, mouse; D059648:Kisspeptins; D049452:Green Fluorescent Proteins