2000 年度実績報告書

新しい脳ペプチド分離の新戦略

研究課題

研究課題/領域番号	11354010
研究機関	国立遺伝学研究所
研究代表者	藤澤敏孝国立遺伝学研究所, 個体遺伝研究系, 助教授 (60000262)
研究分担者	筒井和義広島大学, 総合科学部, 教授 (20163842) 小泉修福岡女子大学, 人間環境学部, 教授 (50094777) 岡戸信男筑波大学, 基礎医学系, 教授 (50060140) 松島治広島大学, 理学部, 教授 (80127711) 藤本正昭島根大学, 生物資源科学部, 教授 (90081728)
キーワード	脳ペプチド / 組織的同定 / フナCRFa / ゴナドトロピン放出抑制ホルモン / マダコGnRH
研究概要	本年度の主な研究実績を以下に列挙する。 1.ヒドラペプチドの組織的分離:約100のペプチドを単離し、50についてアミノ酸配列決定を行った。そのうち10種について化学合成を行い生物活性検定を行っている。 2.ヒドラ神経ペプチドHym-176に対する抗体を用いた齧歯類脳内の類似ペプチドの局在解析と単離:抗体陽性細胞体は視床下部弓状核に、陽性繊維は視床下部室傍核、背内側核、中脳中心灰白質などに存在する。ELISAを用いた抗Hym-176抗体に反応するペプチドを単離し、アミノ酸配列を決定した。現在、このペプチドの詳細な解析を行っている。 3.無脊椎動物のRFアミドに対する抗体を用いた脊椎動物の脳からの類似ペプチドの単離:フナ脳からCRFaを単離し、構造決定した。CRFaはエラによる浸透圧調整に関わる。また、抗CRFa抗体による免疫組織染色からCRFaは嗅球,眼でも発現があり、多機能であることが予想された。CRFaは最近哺乳類脳から単離されたプロラクチン放出ペプチド(PrRP)と相同であり、CRFaにも哺乳類のPrRP受容体と結合し、プロラクチンを放出する活性がある。鳥類脳からはゴナドトロピンの放出を抑制する新規ペプチド(ゴナドトロピン放出抑制ホルモン、GnIHと命名)を同定した。カエルからも同様のペプチドを同定しており、このペプチドが少なくとも脊椎動物に普遍的に存在すると考えられる。 4.無脊椎動物脳からの新規ペプチドの単離:軟体動物、マダコ脳から性腺刺激ホルモン放出ホルモン(GnRH)の同族体と考えられるペプチドを単離同定した。mRNA発現解析、競合ELISA、質量分析、免疫組織染色等からこのペプチドは食道上,下脳、嗅葉-脚葉複合体、視柄腺等に存在する。本ペプチドは神経機能のみならず内分泌ホルモンとしてマダコ生殖機能に関与するGnRHであると考えられる。

研究成果

(36件)

すべてその他

すべて文献書誌 (36件)

[文献書誌] Mochizuki K.: "Universal occurrence of the vasa-related genes among metazoans and their germline expression in Hydra."Development, Genes and Evolution. (In press).
[文献書誌] Bosch T.C.G.: "Polyps, peptides and patterning."Bioessays. (In press).
[文献書誌] Takahashi T.: "Identification of a new member of the Lwamide peptide family : Physiological activity and cellular localization in Cnidaian polyps."Biological Bulletin. (In press).
[文献書誌] Harafuji N.: "Enhancement of foot formation in Hydra by a novel epitheliopeptide, Hym-323."Development. 210. 437-446 (2001)
[文献書誌] Cardenas M.: "Selective protein kinase inhibitors block head specific differentiation in hydra."Cellular Signaling. 12. 649-658 (2000)
[文献書誌] Mochizuki K.: "Expression and evolutionary conservation of nanos-related genes in Hydra."Development, Genes and Evolution. 210. 591-602 (2000)
[文献書誌] Ukena K.: "Developmental changes in progesterone biosynthesis and metabolism in the quail brain."Brain Res.. (In press).
[文献書誌] Matsunaga M.: "Expression and localization of cytochrome P45017 α-hydroxylase/c17,20lyase in the avian brain."Brain Res.. (In press).
[文献書誌] Ukena K: "Distribution of novel RFamide-related peptide-like immunoreactivity in the mouse central nervous system."Neurosci.Lett.,. (In press).
[文献書誌] Satake H.: "Characterization of a cDNA encoding a novel avian hypothalamic neuropeptide exerting an inhibitory effect on gonadotropin release."Biochemical J. (In press).
[文献書誌] Lea R.W.: "Central steroid receptor expression, steroid synthesis and dopaminergic activity related to photoperiod and reproduction in the avian brain."Neurosci.Res.. 36. 261-273 (2000)
[文献書誌] Sakamoto H.: "Existence of galanin in lumbosacral sympathetic ganglionic neurons that project to the quail uterine oviduct."Endocrinology. 141. 4402-4412 (2000)
[文献書誌] Tsutsui K.: "A novel avian hypothalamic peptide inhibiting gonadotropin release."Biochem.Biophys.Res.Commun.. 275. 661-667 (2000)
[文献書誌] Kohchi C.: "Avian galanin : Cloning of complementary DNAs and characterization of transcripts in different tissues."J.Exp.Zool.. 287. 183-190 (2000)
[文献書誌] Tsutsui K.: "Novel cerebellar function : Biosynthesis and actions of neurosteroids in the Purkinje neuron."Int.J.Mol.Med.,. 6. 218-219 (2000)
[文献書誌] Narita N.: "Serotonin transporter gene variation is a risk factor for sudden infant death syndtrome in Japanese population."Pediatrics. (In press).
[文献書誌] Inami S.: "Immunohistochemical demonstration of nerve fibers in the synovial fold of the human cervical facet joint."J Ortho Res.. (In press).
[文献書誌] Okado N.: "A biogenic amine-synapse mechanism for mental retardation and developmental disabilities."Brain Dev.. (In press).
[文献書誌] Maxima M.: "Serotonin 2A receptor-like immunoreactivity is detected in astrocytes, but not in oligodendrocytes"Brain Res.,. 889. 270-273 (2001)
[文献書誌] Shutoh,F: "Long term depletion of serotonin leads to selective changes in glutamate receptor subunits."Neurosci.Res.,. 38. 365-371 (2000)
[文献書誌] Yamaguchi M.: "Time-course effects of a single administration of cocaine on receptor binding and subunit mRNAs of GABA (A) receptors."Mol Brain Res.. 81. 155-163 (2000)
[文献書誌] Masuda,T.: "The involvement of axonin-1/SC2 in mediating notochord-derived chemorepulsive activities for dorsal root ganglion neurites."Dve.Biol.. 224. 112-121 (2000)
[文献書誌] Ikemoto,K.: "Human midbrain dopamine neurons express serotonin 2A receptor : an immunohistochemical demonstration."Brain Res.. 853. 377-380 (2000)
[文献書誌] Asai,D.: "Preparation of the antibody which discriminates the conformations of ligand-bound and ligand-free estrogen receptors : Applications to the quantitative analysis of ligand bindings."Peptide Chemistry. (In press).
[文献書誌] Kawada T.: "Identification of multiple urechistachykinin petides, gene expression, pharmacological activity, and detection using mass spectrometric analyses."Peptides. 21. 1777-1783 (2000)
[文献書誌] Muneoka Y.: "Comparative aspects of invertebrate neuropeptides."Acta Biol Hung.. 51. 111-132 (2000)
[文献書誌] Wang,X.: "Carassius RFamide, a novel FMRFa-related peptide, is produced within the retina and involved in retinal information processing in cyprinid fish."Neurosci.Lett.. 289. 115-118 (2000)
[文献書誌] Csoknya M.: "Distribution of Eisenia tetradecapeptide immunoreactive neurons in the nervous system of earthworms."Acta Biol.Hungarica. 51. 395-408 (2000)
[文献書誌] Wang X.: "Immunohistochemical localization of C-RFamide, FMRF-related peptide, in the brain of the goldfish, Carassius auratus."Zool.Sci.,. 17. 1067-1074 (2000)
[文献書誌] Morishita F.: "NdWFamide : a novel excitatory peptide for the cardiovascular regulation of Aplysia."Peptides. 22. 183-189 (2000)
[文献書誌] Matsushima O.: "Annelid bioactive peptides : A comparative view."Trends in Comp.Biochem.Physiol.. (In press).
[文献書誌] Wang X.: "A candidate for fish prolactin-releasing peptide isolated from Japanese crucian carp."Proc.4th Congr.Asia Oseania Soc.Comp.Endocrinol.. (In press).
[文献書誌] 筒井和義: "小脳における神経ステロイドの合成と作用「ステロイドホルモンと脳科学:性・ストレス・脳をめぐって」"第14回「大学と科学」公開シンポジウム組織委員会編. 8 (2000)
[文献書誌] Tsutsui K.: "Neural Plasticity, Development and Steroid Hormone Action, R.J.Handa et al.(eds.)"CRC Press, U.S.A.(In press).
[文献書誌] Tsutsui K.: "Steroidogenesis in the avian brain. In Avian Endocrinology (A.Dawson and C.M.Chaturvedi, eds)"Journal of Endocrinology Ltd.. 15 (2000)
[文献書誌] Tsutsui K.: "Neuronal neurosteroidogenesis in the cerebellum. Molecular Steroidogenesis, M.Okamoto,Y.Ishimura and H.Nawata (eds.)"Universal Academy Press, Tokyo. 4

2000 年度 実績報告書

新しい脳ペプチド分離の新戦略

研究代表者

藤澤 敏孝 国立遺伝学研究所, 個体遺伝研究系, 助教授 (60000262)

研究成果

[文献書誌] Mochizuki K.: "Universal occurrence of the vasa-related genes among metazoans and their germline expression in Hydra."Development, Genes and Evolution. (In press).

[文献書誌] Bosch T.C.G.: "Polyps, peptides and patterning."Bioessays. (In press).

[文献書誌] Takahashi T.: "Identification of a new member of the Lwamide peptide family : Physiological activity and cellular localization in Cnidaian polyps."Biological Bulletin. (In press).

[文献書誌] Harafuji N.: "Enhancement of foot formation in Hydra by a novel epitheliopeptide, Hym-323."Development. 210. 437-446 (2001)

[文献書誌] Cardenas M.: "Selective protein kinase inhibitors block head specific differentiation in hydra."Cellular Signaling. 12. 649-658 (2000)

[文献書誌] Mochizuki K.: "Expression and evolutionary conservation of nanos-related genes in Hydra."Development, Genes and Evolution. 210. 591-602 (2000)

[文献書誌] Ukena K.: "Developmental changes in progesterone biosynthesis and metabolism in the quail brain."Brain Res.. (In press).

[文献書誌] Matsunaga M.: "Expression and localization of cytochrome P45017 α-hydroxylase/c17,20lyase in the avian brain."Brain Res.. (In press).

[文献書誌] Ukena K: "Distribution of novel RFamide-related peptide-like immunoreactivity in the mouse central nervous system."Neurosci.Lett.,. (In press).

[文献書誌] Satake H.: "Characterization of a cDNA encoding a novel avian hypothalamic neuropeptide exerting an inhibitory effect on gonadotropin release."Biochemical J. (In press).

[文献書誌] Lea R.W.: "Central steroid receptor expression, steroid synthesis and dopaminergic activity related to photoperiod and reproduction in the avian brain."Neurosci.Res.. 36. 261-273 (2000)

[文献書誌] Sakamoto H.: "Existence of galanin in lumbosacral sympathetic ganglionic neurons that project to the quail uterine oviduct."Endocrinology. 141. 4402-4412 (2000)

[文献書誌] Tsutsui K.: "A novel avian hypothalamic peptide inhibiting gonadotropin release."Biochem.Biophys.Res.Commun.. 275. 661-667 (2000)

[文献書誌] Kohchi C.: "Avian galanin : Cloning of complementary DNAs and characterization of transcripts in different tissues."J.Exp.Zool.. 287. 183-190 (2000)

[文献書誌] Tsutsui K.: "Novel cerebellar function : Biosynthesis and actions of neurosteroids in the Purkinje neuron."Int.J.Mol.Med.,. 6. 218-219 (2000)

[文献書誌] Narita N.: "Serotonin transporter gene variation is a risk factor for sudden infant death syndtrome in Japanese population."Pediatrics. (In press).

[文献書誌] Inami S.: "Immunohistochemical demonstration of nerve fibers in the synovial fold of the human cervical facet joint."J Ortho Res.. (In press).

[文献書誌] Okado N.: "A biogenic amine-synapse mechanism for mental retardation and developmental disabilities."Brain Dev.. (In press).

[文献書誌] Maxima M.: "Serotonin 2A receptor-like immunoreactivity is detected in astrocytes, but not in oligodendrocytes"Brain Res.,. 889. 270-273 (2001)

[文献書誌] Shutoh,F: "Long term depletion of serotonin leads to selective changes in glutamate receptor subunits."Neurosci.Res.,. 38. 365-371 (2000)

[文献書誌] Yamaguchi M.: "Time-course effects of a single administration of cocaine on receptor binding and subunit mRNAs of GABA (A) receptors."Mol Brain Res.. 81. 155-163 (2000)

[文献書誌] Masuda,T.: "The involvement of axonin-1/SC2 in mediating notochord-derived chemorepulsive activities for dorsal root ganglion neurites."Dve.Biol.. 224. 112-121 (2000)

[文献書誌] Ikemoto,K.: "Human midbrain dopamine neurons express serotonin 2A receptor : an immunohistochemical demonstration."Brain Res.. 853. 377-380 (2000)

[文献書誌] Asai,D.: "Preparation of the antibody which discriminates the conformations of ligand-bound and ligand-free estrogen receptors : Applications to the quantitative analysis of ligand bindings."Peptide Chemistry. (In press).

[文献書誌] Kawada T.: "Identification of multiple urechistachykinin petides, gene expression, pharmacological activity, and detection using mass spectrometric analyses."Peptides. 21. 1777-1783 (2000)

[文献書誌] Muneoka Y.: "Comparative aspects of invertebrate neuropeptides."Acta Biol Hung.. 51. 111-132 (2000)

[文献書誌] Wang,X.: "Carassius RFamide, a novel FMRFa-related peptide, is produced within the retina and involved in retinal information processing in cyprinid fish."Neurosci.Lett.. 289. 115-118 (2000)

[文献書誌] Csoknya M.: "Distribution of Eisenia tetradecapeptide immunoreactive neurons in the nervous system of earthworms."Acta Biol.Hungarica. 51. 395-408 (2000)

[文献書誌] Wang X.: "Immunohistochemical localization of C-RFamide, FMRF-related peptide, in the brain of the goldfish, Carassius auratus."Zool.Sci.,. 17. 1067-1074 (2000)

[文献書誌] Morishita F.: "NdWFamide : a novel excitatory peptide for the cardiovascular regulation of Aplysia."Peptides. 22. 183-189 (2000)

[文献書誌] Matsushima O.: "Annelid bioactive peptides : A comparative view."Trends in Comp.Biochem.Physiol.. (In press).

[文献書誌] Wang X.: "A candidate for fish prolactin-releasing peptide isolated from Japanese crucian carp."Proc.4th Congr.Asia Oseania Soc.Comp.Endocrinol.. (In press).

[文献書誌] 筒井和義: "小脳における神経ステロイドの合成と作用「ステロイドホルモンと脳科学:性・ストレス・脳をめぐって」"第14回「大学と科学」公開シンポジウム組織委員会編. 8 (2000)

[文献書誌] Tsutsui K.: "Neural Plasticity, Development and Steroid Hormone Action, R.J.Handa et al.(eds.)"CRC Press, U.S.A.(In press).

[文献書誌] Tsutsui K.: "Steroidogenesis in the avian brain. In Avian Endocrinology (A.Dawson and C.M.Chaturvedi, eds)"Journal of Endocrinology Ltd.. 15 (2000)

[文献書誌] Tsutsui K.: "Neuronal neurosteroidogenesis in the cerebellum. Molecular Steroidogenesis, M.Okamoto,Y.Ishimura and H.Nawata (eds.)"Universal Academy Press, Tokyo. 4

2000 年度実績報告書

藤澤敏孝国立遺伝学研究所, 個体遺伝研究系, 助教授 (60000262)