1997 年度実績報告書

新しいカルシウムーリン脂質結合蛋白質、アネキシンVの生理機能に関する研究-視床下部-下垂体-卵巣系をモデルとした細胞生物学的研究-

研究課題

研究課題/領域番号	07456136
研究機関	北里大学
研究代表者	汾陽光盛北里大学, 獣医畜産学部, 助教授 (00153007)
研究分担者	西原真杉東京大学, 農学部, 助教授 (90145673) 長谷川喜久北里大学, 獣医畜産学部, 教授 (40092001) 久留主志朗北里大学, 獣医畜産学部, 助手 (50215076)
キーワード	アネキシンV / 黄体形成ホルモン(LH) / 下垂体 / 遺伝子発現 / プロラクチン / 黄体
研究概要	本研究計画は、アネキシンVが、下垂体、卵巣機能の新たな調節因子である可能性を追求したものである。今年度に得られた結果は以下のように要約される。 1.アネキシンVの遺伝子発現の解析卵巣摘出によって下垂体の性腺刺激ホルモン分泌の亢進しているときに、アネキシンVmRNAの著しく増加することを発見した。この現象は、免疫組織化学的にも、去勢細胞に大量のアネキシンVの出現することで確かめられ、ゴナドトロフの機能亢進にアネキシンVが何らかの役割を担っていることが推定された。そこで、下垂体細胞初代培養系を用いて、LHRHのアネキシンVmRNAに対する、組換えアネキシンVのLHβサブユニットmRNAに対する影響を調べると、アネキシンV合成がLHRHによって促進されること、LHβサブユニット合成がアネキシンVによって促進されることを見出した。 2.In situ hybridization法を用いた、アネキシンV産生細胞の同定 in situ hybridization法を用いて、アネキシンVmRNAの組織内分布を明らかにすることを試みた。卵巣でまず実験を行い、黄体細胞の中でも大型の細胞にmRNAの出現すること、顆粒層細胞にはアネキシンVmRNAの存在しないこと、黄体のアネキシンV合成活性が性周期中に変動すること、ノーザンブロット解析の結果、発情前期の卵巣でアネキシンVmRNAの著増することが明らかとなった。 3.アネキシンVの細胞作用の検討下垂体ホルモン(LHとプロラクチン)の時間分解免疫測定法を確立し、初代培養系で組換えアネキシンV、抗アネキシンV抗体の効果を検討し、アネキシンVがプロラクチン分泌には抑制的にLH分泌には促進的に作用することを明らかにした。特にLH分泌に対する作用は顕著で、アネキシンVが下垂体において少なくともゴナドトロフの機能に関与していることが明らかとなった。

研究成果

(24件)

すべてその他

すべて文献書誌 (24件)

[文献書誌] Noguchi J: "Ontogeny of inhibin secretion in the testis:secretion of inhibin-related proteins from fetal Leydig cells and of bioactive inhibin from Sertoli cells." J.Endocrinology. 155. 27-34 (1997)
[文献書誌] Kaneko H: "Inhibin is involved in the suppression of FSH secretion in the growth phase of the dominantfollicle during the early luteal phase in cows." Domestic Animal Endocrinology. 14(4). 263-271 (1997)
[文献書誌] Madarame H: "Virulent and avirulent Rhodococcus equi infection in T-cell deficient athymic nude mice:pathologic,bacteriologic and immunologic responces." FEMS Immunology and MedicalMicrobiology. 17. 251-262 (1997)
[文献書誌] Hasegawa Y: "Isolation and characterization of inhibin andinhibin-related proteins." J.Reprod.Develop.42(Suppl.). 23-25 (1997)
[文献書誌] Kaneko H: "Follicular growth and FSH secretion incattle." J.Reprod.Develop.42(Suppl.). 37-40 (1997)
[文献書誌] Yoshida S: "A novel steroid inhibitor for a rat 20α-hydroxysteroid dehydrogenase isozyme(HSD1)" Biol.Reprod.57. 1433-1437 (1997)
[文献書誌] Yamanouchi K: "Inhibin α-subunit mRNA is expressed in the corpus luteum of the pregnant mare." J.Reprod.Develop.43. 267-270 (1997)
[文献書誌] Ikeda A: "Different female reproductive phenotypes determined by human growth hormone(hGH)levels in hGH-transgenic rats." Biol.Reprod.56. 847-851 (1997)
[文献書誌] Yamanouchi K: "Equine inhibin-activin βA-subunit mRNA is expressed in the endometrial gland,but not in the trophoblast,during pregnancy." Mol.Repod.Develop.47. 1-7 (1997)
[文献書誌] Ordog T: "On the role of gonadotropin-releasing hormone(GnRH)in the operation of the GnRH pulse generator in the rhesus monkey." Neuroendocrinology. 65. 307-313 (1997)
[文献書誌] 高橋迪雄: "生殖機能と下垂体前葉機能調節系" ヒューマン・サイエンス. 9. 77-85 (1997)
[文献書誌] Yoo MJ: "Involvement of prostaglandins in suppression of gonadotropin-releasing hormone pulse generator activity by tumor necrosis factor-a." J.Reprod.Develop.43. 181-187 (1997)
[文献書誌] Yoo MJ: "Tumor necrosis factor-a mediates endotoxin induced suppression of gonadotropin-releasing hormone pulse generator activity in the rat." Endocrine J.44. 141-148 (1997)
[文献書誌] Kanuka H: "Prolactin expresses differential effect on apoptotic cell death of luteal cells in vivo and in vitro." Endocrine J.44. 11-22 (1997)
[文献書誌] Kurusu S: "Role of cytosolic phospholipase A2 in eicosanoid generation by corpora lutea of pseudopregnant rat:effects of its specific inhibitor" Prostaglandins,Leukotrienes and Essential Fatty Acids. 57(2). 119-124 (1997)
[文献書誌] Kawaminami M: "Immunocytochemical localization of annexin 5,a calcium dependent phospholipid binding protein,in rat endocrine organs" Cell and Tissue Research. (in press.). (1998)
[文献書誌] Suzuki M: "Identification of sex steroid-inducible genes in the hypothalamus of neonatal rats by cDNA subtraction." Neurosci.Lett.(in press.). (1998)
[文献書誌] Bannai M: "Effect of injection of antisense oligonucleotides of GAD isozymes into rat VMH on food intake and locomotor activity." Brain.Res.(in press.). (1998)
[文献書誌] Kawaminami,M: "Ovariectomy enhances the expression of annexin 5 in the anterior pituitary gland and induces nuclear translocation in rat gonadotrophs" Moll.Cell.Endoc.(in press.). (1998)
[文献書誌] Hasegawa Y: "Two-site immunoassay for native inhibin A.in"Inhibin,Activin and Follistatin."" Eds.Aono T,Sugino H and Walww,Springer-Verlag,New York,104-117 (1997)
[文献書誌] Mizunuma H: "Evaluation of maternal serum inhibin as a marker for complicated pregnancy.in“Inhibin,Activin and Follistatin."" Eds.Aono T,Sugino H and Walww,Springer-Verlag,New York,151-161 (1997)
[文献書誌] Nishihara M: "Controlling mechanism of GnRH pulse generator activity in the rat.In:Neural Control of the Reproduction:Physiology and Behavior." Maeda K,Tsukamura H,Yokoyama A(eds)Japan Scientific Societies Press,Tokyo/Karger,Basel,31-55 (1997)
[文献書誌] Suzuki M: "Regulation of the expression of gonadotropin-releasing hormone gene in the hypothalamus.In:Reproductive Biology Update:Novel Tools for Assesment of Environmental Toxicity." Miyamoto H,Manabe N(eds),Nakanishi Printing Co.,Kyoto,261-269 (1998)
[文献書誌] 西原真杉: "脳と生殖-GnRH神経系の進化と適応-生殖の神経内分泌調節" 学会出版センター(印刷中), (1998)

1997 年度 実績報告書

新しいカルシウムーリン脂質結合蛋白質、アネキシンVの生理機能に関する研究-視床下部-下垂体-卵巣系をモデルとした細胞生物学的研究-

研究代表者

汾陽 光盛 北里大学, 獣医畜産学部, 助教授 (00153007)

研究成果

[文献書誌] Noguchi J: "Ontogeny of inhibin secretion in the testis:secretion of inhibin-related proteins from fetal Leydig cells and of bioactive inhibin from Sertoli cells." J.Endocrinology. 155. 27-34 (1997)

[文献書誌] Kaneko H: "Inhibin is involved in the suppression of FSH secretion in the growth phase of the dominantfollicle during the early luteal phase in cows." Domestic Animal Endocrinology. 14(4). 263-271 (1997)

[文献書誌] Madarame H: "Virulent and avirulent Rhodococcus equi infection in T-cell deficient athymic nude mice:pathologic,bacteriologic and immunologic responces." FEMS Immunology and MedicalMicrobiology. 17. 251-262 (1997)

[文献書誌] Hasegawa Y: "Isolation and characterization of inhibin andinhibin-related proteins." J.Reprod.Develop.42(Suppl.). 23-25 (1997)

[文献書誌] Kaneko H: "Follicular growth and FSH secretion incattle." J.Reprod.Develop.42(Suppl.). 37-40 (1997)

[文献書誌] Yoshida S: "A novel steroid inhibitor for a rat 20α-hydroxysteroid dehydrogenase isozyme(HSD1)" Biol.Reprod.57. 1433-1437 (1997)

[文献書誌] Yamanouchi K: "Inhibin α-subunit mRNA is expressed in the corpus luteum of the pregnant mare." J.Reprod.Develop.43. 267-270 (1997)

[文献書誌] Ikeda A: "Different female reproductive phenotypes determined by human growth hormone(hGH)levels in hGH-transgenic rats." Biol.Reprod.56. 847-851 (1997)

[文献書誌] Yamanouchi K: "Equine inhibin-activin βA-subunit mRNA is expressed in the endometrial gland,but not in the trophoblast,during pregnancy." Mol.Repod.Develop.47. 1-7 (1997)

[文献書誌] Ordog T: "On the role of gonadotropin-releasing hormone(GnRH)in the operation of the GnRH pulse generator in the rhesus monkey." Neuroendocrinology. 65. 307-313 (1997)

[文献書誌] 高橋迪雄: "生殖機能と下垂体前葉機能調節系" ヒューマン・サイエンス. 9. 77-85 (1997)

[文献書誌] Yoo MJ: "Involvement of prostaglandins in suppression of gonadotropin-releasing hormone pulse generator activity by tumor necrosis factor-a." J.Reprod.Develop.43. 181-187 (1997)

[文献書誌] Yoo MJ: "Tumor necrosis factor-a mediates endotoxin induced suppression of gonadotropin-releasing hormone pulse generator activity in the rat." Endocrine J.44. 141-148 (1997)

[文献書誌] Kanuka H: "Prolactin expresses differential effect on apoptotic cell death of luteal cells in vivo and in vitro." Endocrine J.44. 11-22 (1997)

[文献書誌] Kurusu S: "Role of cytosolic phospholipase A2 in eicosanoid generation by corpora lutea of pseudopregnant rat:effects of its specific inhibitor" Prostaglandins,Leukotrienes and Essential Fatty Acids. 57(2). 119-124 (1997)

[文献書誌] Kawaminami M: "Immunocytochemical localization of annexin 5,a calcium dependent phospholipid binding protein,in rat endocrine organs" Cell and Tissue Research. (in press.). (1998)

[文献書誌] Suzuki M: "Identification of sex steroid-inducible genes in the hypothalamus of neonatal rats by cDNA subtraction." Neurosci.Lett.(in press.). (1998)

[文献書誌] Bannai M: "Effect of injection of antisense oligonucleotides of GAD isozymes into rat VMH on food intake and locomotor activity." Brain.Res.(in press.). (1998)

[文献書誌] Kawaminami,M: "Ovariectomy enhances the expression of annexin 5 in the anterior pituitary gland and induces nuclear translocation in rat gonadotrophs" Moll.Cell.Endoc.(in press.). (1998)

[文献書誌] Hasegawa Y: "Two-site immunoassay for native inhibin A.in"Inhibin,Activin and Follistatin."" Eds.Aono T,Sugino H and Walww,Springer-Verlag,New York,104-117 (1997)

[文献書誌] Mizunuma H: "Evaluation of maternal serum inhibin as a marker for complicated pregnancy.in“Inhibin,Activin and Follistatin."" Eds.Aono T,Sugino H and Walww,Springer-Verlag,New York,151-161 (1997)

[文献書誌] Nishihara M: "Controlling mechanism of GnRH pulse generator activity in the rat.In:Neural Control of the Reproduction:Physiology and Behavior." Maeda K,Tsukamura H,Yokoyama A(eds)Japan Scientific Societies Press,Tokyo/Karger,Basel,31-55 (1997)

[文献書誌] Suzuki M: "Regulation of the expression of gonadotropin-releasing hormone gene in the hypothalamus.In:Reproductive Biology Update:Novel Tools for Assesment of Environmental Toxicity." Miyamoto H,Manabe N(eds),Nakanishi Printing Co.,Kyoto,261-269 (1998)

[文献書誌] 西原真杉: "脳と生殖-GnRH神経系の進化と適応-生殖の神経内分泌調節" 学会出版センター(印刷中), (1998)

1997 年度実績報告書

汾陽光盛北里大学, 獣医畜産学部, 助教授 (00153007)