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Development of culture system of mammalian circadian oscillator and its molecular analysis

Research Project

Project/Area Number 02454105
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 基礎獣医学
Research InstitutionUniversity of Miyazaki

Principal Investigator

ETOH Teiichi  University of Miyazaki, Professor, 農学部, 教授 (70081510)

Co-Investigator(Kenkyū-buntansha) MURAKAMI Noboru  University of Miyazaki, Assistant Professor, 農学部, 助手 (80150192)
Project Period (FY) 1990 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥4,800,000 (Direct Cost: ¥4,800,000)
Fiscal Year 1992: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1991: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1990: ¥2,600,000 (Direct Cost: ¥2,600,000)
KeywordsBiological clock / Pineal gland / Suprachiasmatic nucleus / Circadian rhythm / サーカディアンリズム / バゾプレッシン / 培養細胞 / 日内リズム / 神経培養
Research Abstract

1. Culture of rat suprachiasmatic nucleus neuron: We have developed a dissociated suprachiasmatic nucleus (SCN) cell culture system in order to begin a cellular analysis of mammalian circadian oscillator. Twenty seven hour oscillation of vasopressin release was observed in SCN cell culture. This finding suggest that cultured SCN neurons retain the capacity for circadian oscillation for a long time, and should provide a useful model to analyze the mammalian clock mechanism.
2. Reentrainment of circadian clock itself to new light-dark cycle: We investigated whether the circadian clock itself reentrain more quickly to a new light-dark cycle than does the overt rhythm of activity.It took 2.4 days and 6.4 days for clock itself and overt activity rhythm to re-entrain to the new light-dark cycle, respectively. This results suggest that the circadian clock itself reentrain very rapidly following change of light-darkcycle, while the overt rhythms of activity take many cycles.
3. Search of proteins related to circadian clock mechanism: We investigated the time-dependent protein in SCN cells harvested every 3 hour from rats kept under constant dark conditions. A 64-kDa protein synthesis rate was rhythmic, and showed two distinct peaks which one of them are observed at around CT 6-9, and the other around CT 21. The peak times consisted with the region of phase-advance induced light and protein synthesis inhibitor.These results indicate the 64kDa protein in SCN may be involved in circadian clock mechanism.
4. We examined the difference of circadian clock mechanism between various species of avian, such as chick, crow, pigeon, owl, sparrow, and quail. The pineal gland of owl did not show the circadian rhythm of melatonin release. On the other hand the other avian, except a quail, showed the circadian oscillation of melatonin release under both LD cycle and DD conditions. These results suggest that circadian clock mechanism of avian pineal is different among the species.

Report

(4 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • 1990 Annual Research Report
  • Research Products

    (15 results)

All Other

All Publications (15 results)

  • [Publications] Murakami,N.Takamure,M.Takahashi,K.Utunomiya,K.Kuroda,H.Etoh,T.: "Long-term culfured neu rone from rat supra chiasmatic nucleus retain the capacity for circadian oscillation of vasopressin release." Brain Research. 545. 347-350 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Takamure,M.Murakami,N.Takahashi,K.Kuroda,H Etoh,T.: "Rapid reentrainment of the circadian clock itself,but not measurable activity rhythms in the rat." Physiology & Behavior. 50. 443-449 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] 村上 昇: "鳥類松果体細胞での時計関連蛋白質の存在" ホルモンと臨床. 39. 11-16 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] 村上 昇,黒田 治門 江藤 禎一: "分離松果体細胞の日周リズム" 生体の科学. 42. 585-588 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] N.Murakami,N.Takamure,K.Takahashi,K.Utunomia,K.Kuroda,T.Etoh: "Longーterm cultured neurones from rat suprachiasmatic nucleus retain the capacity for ciradion oscillation of vasopressin release" Brain Research. 545. 347-350 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] M.Takamure,N.Muradami,K.Takahashi,H.Kuroda,T.Etoh: "Rapid reーentrainment of the circadian clock itself,but not measurable activity rhythms,to a new lightーdark cycle in the rat." Physiology and Behavior. 50. 443-449 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] N.Murakami: "Involvement of protein synthesis in the circadian clock and light entrainment in chich pineal cells." Jpn.J.Anim.Reprod.36. 53-61 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 村上 昇: "鳥類松果体細胞での時計関連蛋白質の存在" ホルモンと臨床. 39. 11-16 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 村上 昇,黒田 治門,江藤 禎一: "分離松果体細胞の日周リズム" 生体の科学. 42. 585-588 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Masato Egawa,shuj.Inoue,Shinobu Sato,Yutaro Takamura,Noboru Murakami,Kiyohisa Takahashi: "Restoration of ciradian corticosterone vhythm in Ventromedial Hypothalamic Lesioned rats." Neuroendocrinology. 53. 543-548 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] N.Murakami,M.Takamure,K.Takahashi,K.Utunomya,H.Kuroda,T.Etoh.: "Longーterm cultured neurones from rat suprachiasmatic nucleus retain the capacity for circadian oscillation of vasopressin release" Brain Research. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] M.Takamure,N.Murakami,K.Takahashi,H.Kuroda,T.Etoh: "Rapid reーentrainment of the circadian clock itself,but not measurable rhythms,to a new lightーdark cycle in the rat." Physiology and Behavior. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] 高橋 清久.大川 匡子.村上 昇: "ほ乳動物のリズム同調機構:特に臨床応用面から" 神経研究の進歩. 34. 892-900 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 村上 昇: "鳥類松果体細胞の時計及び光情報に関わる蛋白合成" 家畜繁殖学雑誌. 35. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] 高橋 清久.村上 昇: "神経科学レビュ-" 医学書院(伊藤正男編集), 314 (1990)

    • Related Report
      1990 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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