1997 Fiscal Year Annual Research Report

動物行動の分子生物学的解析-遺伝的睡眠障害動物作成への挑戦-

Research Project

Project/Area Number	07558108
Research Institution	Osaka Bioscience Institute
Principal Investigator	裏出良博 (財)大阪バイオサイエンス研究所, 第2研究部, 副部長 (10201360)
Co-Investigator(Kenkyū-buntansha)	金岡禧秀 (財)大阪バイオサイエンス研究所, 第2研究部, 研究員 (40271514) 松村人志 (財)大阪バイオサイエンス研究所, 第2研究部, 副部長 (50173886) 早石修 (財)大阪バイオサイエンス研究所, 第2研究部, 所長、部長 (40025507)
Keywords	睡眠 / 覚醒 / プロスタグランジンD2 / プロスタグランジンD合成酵素 / 遺伝子工学 / ノックアウトマウス / トランスジェニックマウス / X線結晶解析
Research Abstract	プロスタグランジン(PG)D2は、ラットおよびヒト脳における主要PGであり、現在までに同定された数十種類の睡眠物質の中で最も強力な睡眠誘発作用を示し、人の睡眠病や肥満細胞増多症の患者の睡眠発作にも関与する。我々は内在性睡眠物質であるPGD2の生合成を遺伝子工学の手法を用いて操作し、遺伝的な睡眠障害動物の作成を試みた。まず、PGD2の生合成反応を触媒する二種類のPGD合成酵素、すなわち中枢神経系や雄性生殖器に局在し生理的睡眠調節の鍵を握ると考えられているリポカリン型酵素と免疫系や消化器・皮膚などの末梢臓器の抗原提示細胞や肥満細胞に分布する造血器型酵素のマウスとヒトのcDNAおよび遺伝子のクローニングを行った。その結果、両酵素のアミノ酸配列には全く相同性が認められず、リポカリン型酵素は各種の分泌蛋白質で構成されるリポカリンファミリーに属する唯一の酵素であり、造血器型酵素は脊椎動物で発見された最初のシグマ型のグルタチオン転移酵素であることを見いだした。従って、両者は異なる起源から進化し、その過程で活性中心アミノ酸の空間的相同性を獲得し同じ触媒機能を果たすようになった「機能的相似」の新しい例であると考えられる。さらに大腸菌を用いて発現させた遺伝子組換え型両酵素の結晶化に成功し、X線結晶解析により造血器型酵素の三次元構造座標を決定した。又、ヒト型の両酵素を大量発現するトランスジェニックマウスおよび造血器型酵素遺伝子を欠損したノックアウトマウスの作製を行い、これら遺伝子変異マウスの睡眠量や睡眠内容の判定を行うマウス用の睡眠バイオアッセイシステムを完成した。

Research Products
(24 results)

All Other

All Publications (24 results)

[Publications] Yoshihiro Urada: "Lipid Mediators Cell Signalling" J.Neurosci.257-273. 12. 257-273 (1995)
[Publications] Makoto Murakami: "Interleukin-4 suppresses c-Kit ligand-induced expression of cytosolic phospholipase A2 and prostaglandin endoperoxide synthase-2,and their roles in separate pathways of eicosanoid synthesis in mouse bone marrow-derived mast sells." Proc.Natl.Acad.Sci.USA. 92. 6107-6111 (1995)
[Publications] Bettina Blodorn: "he major site of mRNA expression for the b-trace protein (prostaglandin D synthase) in human brain." Neurosci.Lett.209. 117-120 (1996)
[Publications] Hiroshi Oda: "Quantitative sandwich immunosorbent assay of human secretory prostaglandin D synthase (b-trace)." Proc.Japan Acad.72. 108-111 (1996)
[Publications] Carsten T.Beuckmann: "Lipocalin-type prostaglandin D synthase (b-trace) is located in pigmented epithelial cells of rat retina and accumulates within interphotoreceptor matrix." J.Neurosci.16. 6119-6124 (1996)
[Publications] Yoshihiro Urade: "Molecular mechanism of sleep regulation by prostaglandin D2. J." Lipid Mediators Cell Signalling. 14. 71-82 (1996)
[Publications] Dimitrios N.Melegos: "Immunofluorometric assay of prostaglandin D synthase (b-trace) and preliminary clinical applications." Clin.Chem.42. 1984-1991 (1996)
[Publications] Hiroyuki Toh: "IGlutathione-independent prostaglandin D synthase as a lead molecule for designing new functional proteins." Protein Engineering. 9. 1067-1082 (1996)
[Publications] Tetsumori Yamashima: "IProstaglandin D synthase (b-trace) in human arachnoid and meningioma cells : roles as a cell marker or in CSF absorption,tumorigenesis,and calcification process." J.Neurosci. 17. 2376-2382 (1997)
[Publications] Toshiki Tanaka: "Lipocalin-type prostaglandin D synthase (b-trace) in a newly recognized type of retinoid transporter." J.Biol.Chem.272. 15789-15795 (1997)
[Publications] Luis F.Garcia-Fernandez: "Hypothyroidism alters the expression of prostaglandin D2 synthase/β-trace in specific areas of the developing rat brain. Eur." J.Neurosci.9. 1566-1573 (1997)
[Publications] Atsushi Hiraoka: "Analysis of low-molecular-mass proteins in cerebrospinal fluid by sodium dodecyl sulfate capillary electrophoresis." J.Chromatogr.697. 141-147 (1997)
[Publications] Peter Dieter: "Arachidonic acid cascade in kupffer cells." Cells Hepatic Sinusoid. 6. 355-356 (1997)
[Publications] Yoshihide Kanaoka: "Cloning and crystal structure of hematopoietic prostaglandin D synthase." Cell. 90. 1085-1095 (1997)
[Publications] Ishtiaq Mahmud: "Prostaglandin D synthase in human megakaryoblastic cells." J.Biol.Chem.272. 28263-28266 (1997)
[Publications] Yutaka Eguchi: "Expression of lipocalin-type prostaglandin D synthase (b-trace) in human heart and its accumulation in the coronary circulation of angina patients." Proc.Natl.Acad.Sci.USA.94. 14689-14694 (1997)
[Publications] Yoshihiro Tokugawa: "Lipocalin-type prostaglandin D synthase in human male reproductive organs and seminal plasma." Biol.Reprod. 58. 600-607 (1998)
[Publications] Yoshihiro Urade: "Gene engineering studies on sleep" Jpn.Clin.Med.56. 488-492 (1998)
[Publications] Yoshihiro Urada: "Gene engineering studies on sleep" Jpn.Clin.Med.56. 488-492 (1998)
[Publications] Atsushi Hiraoka: "Sodium dodecyl sulfate capillary electrophoretic analysis of molecular mass heterogeneity of b-trace protein in cerebrospinal fluid from patients with central nervous system diseases." J.Chromatogr.(in press). (1998)
[Publications] Dmitry Gerashchenko: "Localization of lipocalin-type prostanoid D synthase (b-trace) in iris,cilliary body,and eye fluids." Invest.Ophthalmol.Vis.Sci.(in press). (1998)
[Publications] Robyn L.Gerena: "Identification of a fertility-associated protein in bull seminal plasma as lipocalin-type prostaglandin D synthase." Biol.Reprod.(in press). (1998)
[Publications] Luis F.Garcia-Fernandez: "Identification of a thyroid hormone response element in the 5'-flanking region of the rat lipocalin-type prostaglandin D2 synthase (b-trace) gene." Mol.Brain.Res.(in press). (1998)
[Publications] Dmitry Gerashchenko: "Dominant expression of rat prostanoid DP receptor mRNA in leptomeninges,inner segments of photoreceptor cells,iris epithelium,and cilliary processes." J.Neurochem. (in press). (1998)

1997 Fiscal Year Annual Research Report

動物行動の分子生物学的解析-遺伝的睡眠障害動物作成への挑戦-

Principal Investigator

裏出 良博 (財)大阪バイオサイエンス研究所, 第2研究部, 副部長 (10201360)

Research Products

[Publications] Yoshihiro Urada: "Lipid Mediators Cell Signalling" J.Neurosci.257-273. 12. 257-273 (1995)

[Publications] Bettina Blodorn: "he major site of mRNA expression for the b-trace protein (prostaglandin D synthase) in human brain." Neurosci.Lett.209. 117-120 (1996)

[Publications] Hiroshi Oda: "Quantitative sandwich immunosorbent assay of human secretory prostaglandin D synthase (b-trace)." Proc.Japan Acad.72. 108-111 (1996)

[Publications] Carsten T.Beuckmann: "Lipocalin-type prostaglandin D synthase (b-trace) is located in pigmented epithelial cells of rat retina and accumulates within interphotoreceptor matrix." J.Neurosci.16. 6119-6124 (1996)

[Publications] Yoshihiro Urade: "Molecular mechanism of sleep regulation by prostaglandin D2. J." Lipid Mediators Cell Signalling. 14. 71-82 (1996)

[Publications] Dimitrios N.Melegos: "Immunofluorometric assay of prostaglandin D synthase (b-trace) and preliminary clinical applications." Clin.Chem.42. 1984-1991 (1996)

[Publications] Hiroyuki Toh: "IGlutathione-independent prostaglandin D synthase as a lead molecule for designing new functional proteins." Protein Engineering. 9. 1067-1082 (1996)

[Publications] Tetsumori Yamashima: "IProstaglandin D synthase (b-trace) in human arachnoid and meningioma cells : roles as a cell marker or in CSF absorption,tumorigenesis,and calcification process." J.Neurosci. 17. 2376-2382 (1997)

[Publications] Toshiki Tanaka: "Lipocalin-type prostaglandin D synthase (b-trace) in a newly recognized type of retinoid transporter." J.Biol.Chem.272. 15789-15795 (1997)

[Publications] Luis F.Garcia-Fernandez: "Hypothyroidism alters the expression of prostaglandin D2 synthase/β-trace in specific areas of the developing rat brain. Eur." J.Neurosci.9. 1566-1573 (1997)

[Publications] Atsushi Hiraoka: "Analysis of low-molecular-mass proteins in cerebrospinal fluid by sodium dodecyl sulfate capillary electrophoresis." J.Chromatogr.697. 141-147 (1997)

[Publications] Peter Dieter: "Arachidonic acid cascade in kupffer cells." Cells Hepatic Sinusoid. 6. 355-356 (1997)

[Publications] Yoshihide Kanaoka: "Cloning and crystal structure of hematopoietic prostaglandin D synthase." Cell. 90. 1085-1095 (1997)

[Publications] Ishtiaq Mahmud: "Prostaglandin D synthase in human megakaryoblastic cells." J.Biol.Chem.272. 28263-28266 (1997)

[Publications] Yutaka Eguchi: "Expression of lipocalin-type prostaglandin D synthase (b-trace) in human heart and its accumulation in the coronary circulation of angina patients." Proc.Natl.Acad.Sci.USA.94. 14689-14694 (1997)

[Publications] Yoshihiro Tokugawa: "Lipocalin-type prostaglandin D synthase in human male reproductive organs and seminal plasma." Biol.Reprod. 58. 600-607 (1998)

[Publications] Yoshihiro Urade: "Gene engineering studies on sleep" Jpn.Clin.Med.56. 488-492 (1998)

[Publications] Yoshihiro Urada: "Gene engineering studies on sleep" Jpn.Clin.Med.56. 488-492 (1998)

[Publications] Atsushi Hiraoka: "Sodium dodecyl sulfate capillary electrophoretic analysis of molecular mass heterogeneity of b-trace protein in cerebrospinal fluid from patients with central nervous system diseases." J.Chromatogr.(in press). (1998)

[Publications] Dmitry Gerashchenko: "Localization of lipocalin-type prostanoid D synthase (b-trace) in iris,cilliary body,and eye fluids." Invest.Ophthalmol.Vis.Sci.(in press). (1998)

[Publications] Robyn L.Gerena: "Identification of a fertility-associated protein in bull seminal plasma as lipocalin-type prostaglandin D synthase." Biol.Reprod.(in press). (1998)

[Publications] Luis F.Garcia-Fernandez: "Identification of a thyroid hormone response element in the 5'-flanking region of the rat lipocalin-type prostaglandin D2 synthase (b-trace) gene." Mol.Brain.Res.(in press). (1998)

[Publications] Dmitry Gerashchenko: "Dominant expression of rat prostanoid DP receptor mRNA in leptomeninges,inner segments of photoreceptor cells,iris epithelium,and cilliary processes." J.Neurochem. (in press). (1998)

裏出良博 (財)大阪バイオサイエンス研究所, 第2研究部, 副部長 (10201360)