秒の時間領域から分子進化レベルにわたる生物の光環境応答の解析

1995 Fiscal Year Annual Research Report

• Project/Area Number: 07309020
• Research Category: Grant-in-Aid for Co-operative Research (A)
• Research Institution: The University of Tokyo
• Principal Investigator: 深田 吉孝 東京大学, 大学院・理学系研究科, 教授 (80165258)
• Co-Investigator(Kenkyū-buntansha): 富岡 憲治 山口大学, 理学部, 教授 (30136163) ; 河村 悟 大阪大学, 理学部, 教授 (80138122) ; 津田 基之 姫路工業大学, 理学部, 教授 (60045458) ; 徳永 史生 大阪大学, 理学部, 教授 (80025452) ; 塚原 保夫 東北大学, 大学院・情報科学研究科, 教授 (60004587)
• Keywords: 松果体 / 桿体 / 錐体 / 光受容蛋白質 / G蛋白質 / Caイオン / 概日リズム / 光情報伝達

Research Abstract

交付申請書に記載の研究実施計画に沿って研究を進め、以下の知見を得た。
《秒から分の領域》 桿体型の錐体型の光受容体では、G蛋白質を活性化する中間体の寿命が異なり、光受容体のヘリックス1〜3の数アミノ酸残基の違いにより、この差異が生じることが判明した。つまり、桿体と錐体の光応答の特徴が受容体レベルでかなり説明できた。また、光受容体のリン酸化反応を制御するCa^<2+>結合蛋白質S-モデュリン/リカバリンがロドプシンキナーゼと相互作用して活性を阻害し、視細胞の明暗順応に寄与することが明らかになった。一方、光受容体からG蛋白質への情報変換過程において、G蛋白質の脂質修飾が重要な役割を果たすこと、また脂肪酸構造のみが異なる4種類のGtαは一つの細胞に共存することが示された。同時に、視細胞のcGMP分解酵素の制御サブユニットをリン酸化する酵素を同定し、またタコ網膜よりG蛋白質GiPおよびGqpを単離することに成功した。
《時間から日の領域》 ニワトリ松果体より数種類のG蛋白質αサブユニットの遺伝子クローニングに成功し、概日時計に入力する光情報伝達経路の究明に新たな一歩が踏み出せた。また、カワヤツメ松果体およびマウス網膜において、概日時計へ光信号を入力する光受容体が500nm付近に吸収極大を持つことが示された。一方、コオロギの概日時計は視覚系に影響を与えており、光感度が日周変動すると共に光応答性が昼夜で興奮性から抑制性に逆転する現象が観察された。
《分子進化レベル》 光受容体・G蛋白質なシグナル伝達蛋白質の遺伝子属について分子系統樹を推定・解析した結果、表現系レベルの進化と分子レベルの進化と分子レベルの進化との関連性が強く示唆された。
なお、平成7年12月に2日間にわたって班員全員参加のもとで班会議を開催し、研究成果の報告と活発な討論を行い、今後の研究方針についても共同研究の可能性を含めて綿密に検討した。

Research Products

• [Publications] Daisuke Kojima: "Purification and low temperature spectroscopy of gecko visual pigments green and blue." Biochemistry. 34. 1096-1106 (1995)
• [Publications] Kamon Sanada: "Role of heterogeneous N-terminal acylation of recoverin in rhodopsin phosphorylation." J. Biol. Chem.270. 15459-15462 (1995)
• [Publications] Rika Morishita: "Primary structure of a γ subunit of G protein, γ12, and its phosphorylation by protein kinase C." J. Biol. Chem.270. 29469-29475 (1995)
• [Publications] Toshiyuki Okano: "Molecular basis for tetrachromatic color vision." Comp. Biochem. Physiol.112B. 405-414 (1995)
• [Publications] Daisuke Kojima: "Molecular properties of chimerical of Gecko blue and bovine rhodopsin." Biochemistry. (in press). (1996)
• [Publications] Osamu Shono: "Preparation and characterization of monoclonal antibodies specific for lauroylated isoform of bovine transducin α-subunit." J. Neurochem.(in press). (1996)
• [Publications] 深田吉孝: "生物時計の時刻合わせをする脳内タンパク質" 生物の科学 遺伝. 49. 8-9 (1995)
• [Publications] 深田吉孝: "分子レベルにおける生物時計機構の解析" 生化学. 67. 1137-1142 (1995)
• [Publications] 岡野俊行: "脳は光を感じる「第三の目」" ニュートン. 15. 102-107 (1995)
• [Publications] 深田吉孝: "視物質の多様化と視覚の進化" 細胞工学. 14. 1007-1014 (1995)
• [Publications] 岡野俊行: "光受容タンパク質の分子進化" 生物の科学 遺伝. 49. 28-34 (1995)
• [Publications] 深田吉孝: "光受容体の構造と機能調節" 実験医学. 14. 63-67 (1996)
• [Publications] 真田佳門: "シグナル伝達蛋白質における脂質修飾の役割" 膜. (印刷中). (1996)
• [Publications] Kamon Sanada: "Unique lipids and unique properties of retinal proteins." Behavioral and Brain Sciences. 18. 486-487 (1995)
• [Publications] Tsuda, T.: "Influences of season and of temperature, photoperiod, and subcutaneous melatonin infusion on the glomerular filtration rate of ewes." J. Pineal Res.(in press). (1996)
• [Publications] IIgo, M.: "Circadian rhythms of locomotor activity in the goldfish, Carassius auratus." Physiology and Behavior. (in press). (1996)
• [Publications] Iigo, M.: "Effects of season, temperature , and photoperiod on plasma melatonin rhythms in the goldfish, Carassius auratus." J. Pineal Res.18. 62-68 (1995)
• [Publications] Uchida, K.: "Perioperative plasma melatonin levels in patients with cardiac and esophageal surgery." St. Marianna Medical Journal. 23. 353-360 (1995)
• [Publications] Iigo, M: "Effects of pinealectomy and constant light exposure on day-night changes of melatonin binding sites in the goldfish brain." Neurosci. Lett.197. 61-64 (1995)
• [Publications] Fujisawa, Y.: "Day-night changes in melatonin contents in the vitreous body and in vitro melatonin release from the retina of Japanese quail." St. Marianna Medical Journal. 23. 833-842 (1995)
• [Publications] Yukizane, M., Tomioka, K.: "Neural pathways involved in mutual interactions between optic lobe circadian pacemakers in the cricket Gryllus bimaculatus." Journal of Comparative Physiology A. 176. 601-610 (1995)
• [Publications] Tomioka, K., Agui, N., Bollenbacher, W. E.: "Electrical properties of the cerebral prothoracicotropic hormone cells in diapausing and non-diapausing pupae of the tobacco born worm, Manduca sexta." Zoological Science. 12. 165-173 (1995)
• [Publications] Murata, T., Matsumoto, A., Tomioka, K., Chiba, Y.: "Ritsu : a clock mutant from natural population of Drosophila melanogaster." Journal of Neurogenetics. 9. 239-249 (1995)
• [Publications] 富岡憲治: "コオロギの体内時計" 昆虫と自然. 30. 23-28 (1995)
• [Publications] Araki, M.: "High potassium promotes differentiation of retinal neurous but does not favor rod differentiation" Developmantal Brain Research. 89. 103-114 (1995)
• [Publications] 荒木正介: "神経の発生における分化の多能性とその制御" 京都府立医科大学雑誌. 104. 233-247 (1995)
• [Publications] 荒木正介: "視細胞の分化と細胞メカニズム" 電子顕微鏡. 30. 4-12 (1995)
• [Publications] 荒木正介: "ラット網膜の培養と神経細胞・視細胞の分化" 神経眼科. 12. 261-265 (1995)
• [Publications] Kishigami, A.: "The traffic of particles in the axonic process of vertebrae cone-type photoreceptor cells." Zool. Sci.12. 349-350 (1995)
• [Publications] Suzuki, T.: "Squid photoreceptor phospholipase C is stimurated by membrane Gq but not by soluble Gq" FEBS Letters. 377. 333-337 (1995)
• [Publications] De Castro, E: "Regulation of rhodopsin phosphorylation by a family of neuronal calcium sensors." Biochem. Biophys. Res. Commun.216. 133-140 (1995)
• [Publications] 津田基之: "ものが見えるとはどういうことか 視覚初期過程分子メカニズム" パリティー. 3. 8-15 (1996)
• [Publications] Kuma, K., Nikoh, N., Iwabe, N., and Miyata, T.: "Phylogenetic position of Dictyostelium inferred from mulitple protein data sets." J. Mol. Evol.41. 238-246 (1995)
• [Publications] Zhang, R., Yatsuki, H., Kusakabe, T., Iwabe, N., Miyata, T., Imai, T., Yoshida, M., and Hori, K.: "Structures of cDNAs encoding the muscle-type and non-muscle-type isozyme of lampley fructose bisphosphate aldolases and the evolution of aldolase genes." J. Biochem.177. 545-553 (1995)
• [Publications] Iwabe, N., Kuma, K., Nikoh, N., and Miyata, T.: "Molecular clock for dating of divergence between animal phyla." Jpn. J. Genet.70. 687-792 (1995)
• [Publications] Iwabe, N., Kuma, K., and Miyata, T.: "Evolution of gene families and relationship with organismal evolution : rapid divergence of tissue-specific genes in the early evolution of chordates." Mol. Biol. Evol.13 (in press). (1996)
• [Publications] 岩部直之、宮田隆: "膜電位依存性イオンチャンネルの分子進化" 蛋白質核酸酵素. 40. 2311-2318 (1995)
• [Publications] 宮田隆、岩部直之、隅啓一、菅裕、二河成男: "真核生物遺伝子の断続的多様化と生物進化との関連" 比較生理生化学. 12. 378-389 (1995)
• [Publications] Teraktia A.: "Light-modulated subcellular localization of the alpha-subunit of GTP-binding protein Gq in caryfish photoreceptors." Visual Neuroscience. (in press). (1996)
• [Publications] Tamotsu S.: "Norepinephrine-induced phosphorylation of the transcription factor CREB in isolated rat pinealocytes : an immunocytochemical study." Cell and Tissue Research. 282. 219-226 (1995)
• [Publications] H. Sano et al.: "Effects of suprachiasmatic lesions on circadian rhythms of blood pressure, heart rate and locomotor activity in the rat." Japanese Circulation Journal. 59. 565-573 (1995)
• [Publications] A. Adachi, M. Hasegawa and S. Ebihara: "Measurement of circadian rhythms of ocular melatonin in the pigeon by in vivo microdialysis." NeuroReport. 7 (in press). (1995)
• [Publications] T. Yoshimura and S. Ebihara: "Spectral sensitivity of photoreceptors mediating phase-shifts of circadian rhythms in retinally degenerate CBA/J (rd/rd)・・・" Journal of Comparative Physiology A. 178 (in press). (1996)
• [Publications] Sasaki, M., Masuda, A., Oishi, T.: "Circadian rhythms of corneal mitotic rate, retinal melatonin and immunoreactive visual pigments and the effects of melatonin in the rhythms on the rhythms in Japanses quail." Journal of Comparative Physiology. 176. 465-471 (1995)
• [Publications] Masuda, A., Oishi, T.: "Effects of restricted feeding on the light-induced body weight change and locomotor activity in the Djungarian hamster." Physiology and Behavior. 58. 153-159 (1995)
• [Publications] Oishi, T., Mohri, Y., Kaneko, T. Masuda, A.: "Effects of eyelid suture on the circadian ocular rhythms in Japanese quail." Photomedicine and Photobiology. 17. 41-42 (1995)
• [Publications] Naruse, M., Oishi, T.: "Annual and daily activity rhythms of loaches in an irrigation creek and ditches around paddy fields." Environmental Biology of Fishes. (in press). (1996)
• [Publications] Ikebe, Y., Oishi, T.: "Correlation between environmental parameters and behavior during high tides in the mudskipper, Periophthalmus modetus." Journal of Fish Biology. (in press). (1996)
• [Publications] Hirunagi, K., Kiyoshi, K., Sakakibara, K., Adachi, A., Ebihara, S., Silver, R.: "Opsin-and VIP-like immunoreactive cerebrospinal fluid (CSF)-contacting neurons in the lateral septum of the pigeon." Fourth IBRO World Congress of Neuroscience ABSTRACTS, Rapid Communications, Oxford, New York. 276- (1995)
• [Publications] Hirunagi, K. Rommel, E., Korf, H. -W.: "Ultrastructure of cerebrospinal fluid-contacing neurons immunoreactive to vasoactive intestinal peptide and properties of the blood-brain barrier in the lateral septal organ of the duck." Cell Tissue Res. 279. 123-133 (1995)
• [Publications] H. Imai: "Difference in molecular properties between chicken green and rhodopsin." Biochemistry. 34. 10525-10531 (1995)
• [Publications] S. Tachibanaki: "Effect of chloride on the thermal reverse reaction of intermediate of iodopsin." Biochemistry. 34. 13170-13175 (1995)
• [Publications] 七田芳則: "ロドプシンの超高速異性化反応はどこまで解明されたか。" レーザー研究. 23. 945-953 (1995)
• [Publications] Seiya Kayada: "Cloning and expression of rhodopsin cDNA" Comp. Biochem. Physiol.110B. 599-604 (1995)
• [Publications] Osamu Hisatomi: "Molecular cloning and characterization of the purtative ultraviolet-sensitive visual pigment of goldfish." Vision Res.20 (in press). (1995)
• [Publications] 徳永史生: "動物の色覚" 科学. 65. 485-492 (1995)
• [Publications] 深田吉孝: "ピノプシンによる光シグナル伝達と生物時計.Newメディカルサイエンス『細胞内シグナル伝達のしくみ』" 羊土社(印刷中), (1996)
• [Publications] 深田吉孝: "ニワトリ錐体および桿体視物質の抽出と精製.生物薬科科学実験講座 9巻『ホルモン・生理活性物質II』" 廣川書店, 502 (pp. 378-395) (1996)
• [Publications] 佐藤愛子、利島保 大石正、井深信男(編): "光と人間の生活ハンドブック" 朝倉書店, 373 (1995)
• [Publications] 富岡憲治: "時間を知る生物" 裳華房(印刷中), (1996)
• [Publications] 津田基之: "生体膜-生命の基本形を形づくるもの" 吉岡書店(印刷中), (1996)
• [Publications] 徳永史生: "眼科学大系 10A 眼の発生と遺伝" 中山書店, 250 (1995)
• [Publications] 徳永史生: "分子生理学ノート" 学会出版センター, 227 (1995)
• [Publications] Tomioka, K.: "Neural basis for mutual interaction between circadian pacemakers in the cricket, In : Nervous Systems and Behaviour, eds. M. Burrows et al." Georg Thieme Verlag, Thieme Medical Publishers, Inc., New York, 131 (1995)
• [Publications] 河村悟: "分子神経科学の最先端(分担執筆)視覚中枢の分子機構-網膜-" 厚生社, 342 (P135-P145) (1995)
• [Publications] Satoru Kawamura: "Neurobiology of the vertebrate outer retina 分担 Phototransduction, excitation and adaptation." Chapman & Hall, London, 501 (P105-P131) (1995)