Molecular Biology of a Neuronal Circadian Oscillator

本申请中描述的拟议研究涉及理解24小时（昼夜节律）周期基础的化学机制。昼夜节律周期在植物和动物王国中很常见，并组织了许多重要的过程，包括人类的睡眠和清醒。产生如此长周期的生化机制尚不清楚。调查将使用一个模型系统，即海洋软体动物的眼睛。在恒定的黑暗和温度条件下，当与动物的其他部分分离时，这只眼睛具有表达神经脉冲频率昼夜节律的所有机制。首席研究员将使用生化和生理学方法来生成有关神经元昼夜周期机制的数据。主要的方法将是研究在昼夜周期中特定蛋白质出现的时间。从这些蛋白质的氨基酸序列中，首席研究员将合成寡核苷酸探针，该探针可用于克隆编码这些蛋白质的基因信息。这些研究将开始让我们了解哪些基因和蛋白质在昼夜节律周期中被特异性表达。

The proposed research described in this application is concerned with understanding the chemical machinery that underlies 24-hour (circadian) cycles. Circadian cycles are common in the plant and animal kingdom and organize a number of important processes including sleep and waking in humans. The biochemical mechanisms that generate such a long cycle are not known. The investigations will use a model system, the eye of a marine mollusc Aplysia. This eye has all the machinery for expressing many cycles of a circadian rhythm of nerve impulse frequency, under constant conditions of darkness and temperature, when isolated from the rest of the animal. The principal investigator will use biochemical and physiological methods to generate data about the mechanisms of this neuronal circadian cycle. The major approach will be to study the timing of appearance of particular proteins during the circadian cycle. From the amino acid sequence of these proteins the principal investigator will synthesize oligonucleotide probes which can be used to clone the gene messages that code for these proteins. These studies will begin to us what genes and proteins are being expressed specifically during the circadian cycle.