Search of effective agents for biological clock and analysis for the mechanism on circadian clock using cultured neuron

利用培养神经元寻找生物钟有效药物并分析生物钟机制

• 批准号: 04660326
• 负责人: MURAKAMI Nobuo
• 金额: $ 1.41万
• 依托单位: University og Miyazaki
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04660326/
• 关键词: Biological Clock; Pineal Gland; Suprachiasmatic; Circadian rhythm; メラトニン; 視交叉上核; 概日日内リズム; サーカディアンリズム

1. In cell suspensions of suprachiasmatic nucleus harvested every 3 hr from rats kept under 12 hr : 12hr light-dark cycle and constant darkness, we have detected a Mr 64-kDa protein whose synthesis exhibits two distinct daily peaks in SDS-PAGE.Analysis of densitometer tracings revealed that the synthesis of other proteins was independent of the time of day or not reproducible. Maximum synthesis of the 64-kDa polypeptide occured at around CT6 and CT21, which are almost coincident with the phase advance regions of circadian activity rhythm induced by anisomycin and light pulses [15], respectively. These results suggested that the 64-kDa protein in SDS-PAGE may be a part of the circadian clock mechanism.2. The avian pineal gland plays an important role in the circadian rhythms of behavior and physiology. However, effects of pinealectomy on circadian rhythms differ widely among species of passerine, gallinaceous or columbine birds. We compared the circadian oscillation of melatonin release … More from cultured pineal cells in japanese quail, pigeon and house sparrow, to determine whether the pineal gland of these species retains the circadian oscillator function in vitro.. After dissociated pineal cells has been cultured for 4 days under 12 h : 12 h light-dark (LD) cycle, they were perfused at a flow rate of 0.25 ml/h for 6-7 days under LD or constant darkness (DD). Melatonin release increased during the dark period and low during the light period in all pineral cell cultures. Under DD conditions, the circadian rhythm of melatonin release persisted for up to 3-4 cycles in pigeon and house sparrow pineal cells. However, the amplitude of the rhythm decreased gradually. However, in Japanese quail pineal cell culture the circadian oscillation of melatonin release was abolished under DD conditions.These results strengthen the argument that the avian pineal gland's role in circadian organization differs between species. The direct demonstration of species-specific differences in the mechanism of the circadian oscillation of melatonin release from pineal cells should provide a useful model for the analysis of the pineal's circadian system at the cellular level. Less

1.在12小时：12小时光暗周期和恒定黑暗条件下，每隔3小时收集大鼠视交叉上核细胞悬液，我们检测到一种分子量为64-kDa的蛋白质，其合成在SDS-PAGE上呈现两个明显的日峰，光密度计示踪分析表明，其他蛋白质的合成与时间无关或不可重复。64-kDa多肽的最大合成发生在CT6和CT 21附近，这分别与茴香霉素和光脉冲诱导的昼夜活动节律的相位提前区域几乎一致[15]。这些结果表明，SDS-PAGE中的64-kDa蛋白可能是生物钟机制的一部分.鸟类松果体在行为和生理的昼夜节律中发挥着重要作用。然而，松果体切除术对昼夜节律的影响在雀形目、鸡目或鸽属鸟类中有很大差异。我们比较了褪黑激素释放的昼夜振荡 ...更多信息 从日本鹌鹑、鸽子和家雀的培养松果体细胞中，以确定这些物种的松果体是否在体外保留昼夜节律振荡器功能。将分离的松果体细胞在12 h：12 h光暗（LD）循环下培养4 d后，在LD或恒暗（DD）条件下以0.25 ml/h的流速灌注6-7 d。褪黑激素的释放增加，在黑暗期间和低的光照期间在所有的松树细胞培养。在DD条件下，褪黑激素释放的昼夜节律在鸽子和家雀松果体细胞中持续长达3-4个周期。但节律的振幅逐渐降低。然而，在日本鹌鹑松果体细胞培养的昼夜节律振荡的褪黑激素释放被废除DD conditions.These结果加强的论点，鸟类松果体的昼夜节律组织的作用不同物种之间。松果体细胞褪黑激素释放的昼夜节律振荡机制的直接示范的物种特异性差异，应提供一个有用的模型，在细胞水平上的松果体的昼夜节律系统的分析。少

项目成果

M.Taniguchi.: "Melatonin release from pineal cells of diurnal and nocturnal birds." Brain Research. 620. 297-300 (1993)

M.Taniguchi.：“白天和夜间鸟类的松果体细胞释放褪黑激素。”

R.Nishi.: "Circadian oscillation of 64KDa polypeptide in the rat suprachiasmatic nucleus." J.Vet.Med.Sci.56. 324-328 (1994)

R.Nishi.：“大鼠视交叉上核中 64KDa 多肽的昼夜节律振荡。”

R.Nishi.: "Circadian oscillation of 64KDa polypeptide in the rat suprachiasmatic nucleus." J.Vet.Med.Sci.56. 321-324 (1994)

R.Nishi.：“大鼠视交叉上核中 64KDa 多肽的昼夜节律振荡。”

M.Taniguchi.: "Melatonin release from pineal cells of diurnal and nocturnal birds" Brain Research. 620. 297-300 (1993)

M.Taniguchi.：“白天和夜间鸟类的松果体细胞释放褪黑激素”大脑研究。

M.Taniguchi., N.Murakami., H.Nakamura., T.Nasu., S.Shinohara and T.Etoh: "Melatonin release from pineal cells of diurnal and nocturnal birds." Brain Research. 620. 297-300 (1993)

M.Taniguchi.、N.Murakami.、H.Nakamura.、T.Nasu.、S.Shinohara 和 T.Etoh：“白天和夜间鸟类的松果体细胞释放褪黑激素。”