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

基本信息

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

项目摘要

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。在大鼠的细胞悬浮液中每3小时收获的细胞悬浮液，保持在12小时以下：12小时的灯光和恒定的dave，我们调用了一种MR 64-kDa蛋白，其合成在SDS-PAGE中表现出两个不同的每日峰。密度计的迹象表明，在CT6和CT21左右，在一天中或不重现的其他蛋白质中合成64 kDa多肽。 15]这些结果表明，SDS-PAGE中的64 kDa蛋白可能是昼夜节律的一部分。2。在哥伦比亚鸟类的红色鸟类中广泛地释放昼夜节律释放……更多来自日本Quail，Pigeon和House Sparrow的培养细胞，以确定圆形圆锥形的圆锥形蛋白的刺激蛋白振荡器振荡器是否偶然。在LD或恒定的黑暗条件下，将其培养为4天的浅黑色（LD）周期，以0.25 ml/h的流速为0.25 ml/h。对于鸽子和室内麻雀细胞中最多3-4个周期，节奏的幅度降低了循环的循环表明特定差异差异差异素从佩内尔细胞中释放出褪黑激素的释放，这是分析细胞水平松果体昼夜节律系统的有用模型。