鳥類の性決定分子機構

1999 Fiscal Year Annual Research Report

• Project/Area Number: 10044201
• Research Category: Grant-in-Aid for Scientific Research (A)
• Research Institution: Nagoya University
• Principal Investigator: 島田 清司 名古屋大学, 大学院・生命農学研究科, 教授 (40065579)
• Co-Investigator(Kenkyū-buntansha): 齋藤 昇 名古屋大学, 大学院・生命農学研究科, 助手 (40211924) ; 朴 民根 東京大学, 大学院・理学系研究科, 助教授 (00228694) ; 友金 弘 日本獣医畜産大学, 獣医畜産学部, 教授 (30023493) ; 小野 珠乙 信州大学, 農学部, 助教授 (10177264) ; 原田 信広 藤田保健衛生大学, 医学部, 教授 (00189705)
• Keywords: ニワトリ / SRC-1 / RT-PCR / アロマターゼ / クローニング / アンチセンスオリゴヌクレオチド / 生殖腺 / SOX9

Research Abstract

鳥類の性は、遺伝的に受精の時点で決定される。すなわち、性染色体の対合がホモ(ZZ)の個体は雄に、ヘテロ(ZW)の個体は雌に決められ、それぞれの生殖腺は精巣から卵巣に誘導される。しかしながら、性の決定に関する遺伝子発現と性分化の機序については不明な点が多い。SOX9という遺伝子が性分化に関係する遺伝子として考えられており、特に雄の性分化に関与することが哺乳類で示唆されている。ニワトリにおけるSOX9の役割を明らかにするために、SOX9アンチセンスオリゴヌクレオチドを孵卵9日胚に投与すると、SOX9のmRNA量の低下が観察された。今後、さらに他の遺伝子発現などを調べ、SOX9の性分化に対する役割を調べる。
私たちはP450aromatase(P450arom)が性分化に重要な役割をしており、雌の生殖腺におけるP450aromの発現は雄に比べ高いことを示した。初期胚にアロマターゼインヒビターを投与すると遺伝的に雌の鶏が雄に性転換することを示した。このP450aromの遺伝子発現がどのようにして調節されているか明らかではない。そして、P450aromの遺伝子発現調節に関与すると考えられる因子の一つであるsteroid receptor coactivator-1(SRC-1)をクローニングし塩基配列を決定した。SRC-1のクローニングは、ヒトのSRC-1から設計したプライマーを用いてRT-PCRにより行い、約1.0kbの塩基配列を決定した。今後、SRC-1の初期胚での遺伝子発現を調べる予定である。

Research Products

• [Publications] 島田清司: "雄と雌が決まる仕組み;魚から鳥、哺乳類まで"生命誌. 7. 4-9 (1999)
• [Publications] Bungo T.: "Effects of naloxone on the anorexic action by glucagon-like peptide-1(7-36) in the neonatal chick."Japanese Poultry Science. 36. 109-115 (1999)
• [Publications] Furuse M.: "Central gastrin inhibits feeding behavior and food passage in neonatal chicks."Life Sciences. 65. 305-311 (1999)
• [Publications] Bungo T.: "Intraventricular injection of glucagon-like peptide-1 inhibits water intake of the neonatal chick"Japanese Poultry Science. 36. 229-235 (1999)
• [Publications] Saito N.: "Gene expression of arginin vasotocin in ovarian and uterine tissues of the chicken."Asian-Australasian Journal of Animal Science. 12. 695-701 (1999)
• [Publications] Saito N.: "Effects of naloxone on neurohypophysial peptides release by hypertonic stimulation in chicks."General and Comparative Endocrinology. 115. 228-235 (1999)
• [Publications] Bungo T.: "Influence of intracerebroventricular administration of morphine and kyotrophin on food intake of the neonatal chick."Japanese Poultry Science. 36. 319-324 (1999)
• [Publications] Ieda T.: "Effect of low calcium diet on messenger ribonucleic acid levels of calbindin-D28K of intestine and shell gland in laying hens in relation to egg shell quality."Japanese Poultry Science. 36. 295-303 (1999)
• [Publications] Bungo T.: "Effects of substitution of N-terminal amino acid of glucagon-like peptide-1(7-36) amide on food intake of the neonatal chick"Life Sciences. 25. PL295-PL299 (1999)
• [Publications] Bungo T.: "Intracerebroventricularly administration of glucagon-like peptide-1 induces sleep-like behavior in the neonatal chick."Japanese Poultry Science. 36. 377-381 (1999)
• [Publications] Kansaku N.: "Expression of messenger ribonucleic acids of luteinizing hormone receptor, P450 17α-hydroxylase and P450 aromatase in isolated granulosa, theca interna and theca externa layers of chicken ovarian follicles during follicular growth"Animal Science Journal. 70. 12-17 (1999)
• [Publications] Bungo T.: "Effects of substitution of N-terminal amino acid of glucagon-like peptide-1(7-36) amide on food intake of the neonatal chick"Life Sciences. 25. PL295-PL299 (1999)
• [Publications] Diano S.: "Aromatase and estrogen receptor immunoreactivity in the coronary arteries of monkeys and human subjects."Menopause. 6. 21-28 (1999)
• [Publications] Utsumi T.: "Steroid sulfatase expression is an independent predictor of reccurrence in human breast cancer."Cancer Res.. 59. 377-381 (1999)
• [Publications] Harada N.: "Aromatase inhibitors and enzyme stability."Endocrine-Related Cancer. 6. 211-218 (1999)
• [Publications] Honda S.: "Identification of cis-acting elements in the proximal promoter region for the brain specific exon 1 of the mouse aromatase gene."Mol.Brain Res.. 66. 122-132 (1999)
• [Publications] Sasano H.: "Aromatase and sex steroid receptors in human vena cava."Endocr.J.. 46. 233-242 (1999)
• [Publications] Harada N.: "Localized expression of aromatase in human vascular tissues."Circ.Res.. 84. 1285-1291 (1999)
• [Publications] Kongo M.: "An association between lipid peroxidation and a-naphthylisothiocyanate-induced liver injury in rats."Toxicology Lett.. 105. 103-110 (1999)
• [Publications] Utsumi T.: "Significance of steroid sulfatase expression in human breast cancer."Breast Cancer. 6. 298-300 (1999)
• [Publications] Harada, N.: "Aromatase and intracrinology of estrogen in hormone-dependent tumors."Oncology. 57. 7-15 (1999)
• [Publications] Yamada,K.: "A GCM motif protein is involved in placenta-specific expression of human aromatase gene."J.Biol.Chem.. 274. 32279-32286 (1999)
• [Publications] Deladoey J.: "Aromatase deficiency caused by a novel P450arom gene mutation: Impact of absent estrogen production on serum gonadotropin concentration in a body."J.Clin.Endocrinol.Metab.. 84. 4050-4054 (1999)
• [Publications] Mano,T.: "Accelerated conversion of dehydroepiandrosterone sulfate to estrogen in a patient with Crow-Fukase syndrome and diabetes mellitus."Endocr.Res.. 25. 371-380 (1999)
• [Publications] Tanaka,M.: "cDNA cloning and developmental alterations in gene expression of the two Pit-1/GHF-1 transcription factors in the chicken pituitary."Gen.Comp.Endocrinol.. 114. 441-448 (1999)
• [Publications] Tsukada,A.: "Effects of thyroid hormone on serum insulin like growth factor binding proteins (IGFBPs) and IGFBP-2 mRNA expression in chickens."Jpn.Poult.Sci.. 36. 155-166 (1999)
• [Publications] Tsutsui K.: "Neurosteroids in vertebrate brains."Comp.Biochem.Physiol.. C124. 121-129 (1999)
• [Publications] 筒井和義: "脳とニューステロイド"医学のあゆみ. 別冊Ver1. 75-77 (1999)
• [Publications] 筒井和義: "脳がステロイドを合成する"ファルマシア. 35. 112-117 (1999)
• [Publications] 筒井和義: "鳥類の微小脳の繁殖戦略-産卵誘起ペプチドとその作用機構"比較生理生化学. 16. 191-199 (1999)
• [Publications] Tsutsui K.: "Production and function of neurosteroids in the cerebellar Purkinje neuron."Internat.J.Mol.Medicine. 4. 49-56 (1999)
• [Publications] Ukena K.: "Expression and activity of of 3 β-hydroxysteroid dehydrogenase/△5-△4-isomerase in different regions of the avian brain."Brain.Res.. 818. 536-542 (1999)
• [Publications] Takase M.: "Pregnenolone, pregnenolone sulfate and cytochrome P450 side-chain cleavage (P450scc) in the amphibian brain and their seasonal changes."Endocrinology. 140. 1936-1944 (1999)
• [Publications] 筒井和義: "脳で新しく見出されたニューロステロイドの合成と作用"Hormone Frontier in Gynecology. 6. 1-6 (1999)
• [Publications] Ukena K.: "Expression and activity of 3 β-hydroxysteroid dehydrogenase/△5-△4-isomerase in the rat Purkinje neuron during neonatal life."Endocrinology. 140. 805-813 (1999)
• [Publications] Kogo,H.: "Gonadotropin-releasing hormone receptor mRNA expression in the ovaries of neonatal and adult rats."Cells Tissues Organs. 164. 14-22 (1999)
• [Publications] 友金 弘: "猫を科学する (9) III. 猫の繁殖 (1)"畜産の研究. 53. 523-527 (1999)
• [Publications] 友金 弘: "猫を科学する (10)III. 猫の繁殖 (2)"畜産の研究. 53. 625-629 (1999)
• [Publications] 友金 弘: "猫を科学する (11)III. 猫の繁殖 (3)"畜産の研究. 53. 717-720 (1999)
• [Publications] 友金 弘: "猫を科学する (12)III. 猫の繁殖 (4)"畜産の研究. 53. 823-826 (1999)
• [Publications] Utsumi T.: "Correlation of cyclin D1 mRNA levels with clinicopathological parameters and clinical outcome in human breast carcinoma."Int.J.Cancer. (in press). (2000)
• [Publications] Tsutsi K.: "Novel brain function: biosynthesis and actions of neurosteroids in neurons."Neurosci.Res.. (in press). (2000)
• [Publications] Tan F.-I: "Molecular cloning and functional characterization of a vasotocin receptor subtype that is expressed in the shell gland and brain of the domestic chicken."Biology of Reproduction. 62. 8-15 (2000)
• [Publications] Nishikimi H.: "Sex differentiation and mRNA expression of P450c17, P450arom and AMH in gonads of the chicken."Molecular Reproduction and Development. 55. 20-30 (2000)
• [Publications] Suzuki T.: "Cytogenetical assignment of 29 functional genes on chicken microchromosomes by FISH."Cytogenetics Cell Genet.. (in press). (2000)
• [Publications] Suzuki T.: "Comparative FISH mapping on Z chromosomes of chicken and Japanese quail"Cytogenetics Cell Genet.. 87. 22-26 (1999)
• [Publications] Sasaki T.: "Regulation of opioid peptides on the release of arginine vasotocin in the hen."Journal of Experimental Zoology. (in press). (2000)
• [Publications] Ieda T.: "Changes in parathyroid hormone related peptide receptor binding in the shell gland of laying hens (Gallus domesticus) during the oviposition cycle."General and Comparative Endocrinology. (in press). (2000)
• [Publications] Suzuki T.: "Cytogenetical mapping of thirty-one functional genes on chicken chromosomes by the direct R-banding FISH."Cytogenetics Cell Genet.. 87. 32-40 (1999)
• [Publications] Kikuchi M.: "Recent Progress in Molecular and Comparative Endocrinology"Hormone Research Center. 625 (1999)
• [Publications] Kikuchi M.: "Recent Progress in Molecular and Comparative Endocrinology"Hormone Research Center. 625 (1999)
• [Publications] Kikuchi M.: "Recent Progress in Molecular and Comparative Endocrinology"Hormone Research Center. 625 (1999)
• [Publications] Ono,T.: "Developmental Biology Protocols.Vol.I"Humana Press Inc,. 616 (1999)
• [Publications] Park,M.K.: "Recent Progress in Molecular and Comparative Endocrinology"Hormone Research Center. (1999)
• [Publications] 友金 弘: "哺乳類の生殖生物学"学窓社. (1999)