BIOLOGICAL CLOCKS--GENES, BEHAVIOR AND NEUROBIOLOGY

生物钟——基因、行为和神经生物学

• 批准号: 6271745
• 负责人: JEFFREY C HALL
• 金额: $ 17.1万
• 依托单位: BRANDEIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6271745
• 关键词: Drosophilidae; behavioral /social science research tag; behavioral genetics; biological clocks; biological signal transduction; chimeric proteins; circadian rhythms; cyclic AMP; developmental genetics; developmental neurobiology; ethology; fusion gene; gene expression; gene mutation; genetically modified animals; immunocytochemistry; laboratory rabbit; molecular cloning; molecular genetics; neuroanatomy; neurohormones; photobiology; site directed mutagenesis

Circadian rhythms are organismically ubiquitous biological cycles whose propr daily regulation is strongly connected to the well being of species ranging from microbes to mammals. The 25- year-old genetic approach toward understanding these rhythms has recently begun to reveal elements of the clock mechanisms that underlie daily rhythmicities. This sub-proposal in a Program Project application revolves around the behavioral genetic, neuro- genetics, and molecular neurobiology of circadian rhythms in Drosophila. The experiments proposed stress more of a ~systems~ approach than one that would concentrate upon the hard core of molecular pacemaking. Thus, the neural substrates of behavioral rhythms, and a periodic feature of late development, will be delved into by application of rhythm mutants and transgenic strains carrying manipulated forms of clock-genes; these studies include descriptions of anatomic output pathways from neurons that are candidate for CNS-pacemaker cells, and selectively effected perturbations of the structure and function of such cells in conjunction with bioassaying the effects of the molecularly mediated neuronal damage. Input paths to the central pacemakers, which bring in environmental cues to effect crucial daily re-sets of the clock, will be dissected both in terms of anatomy and elements of signal-transduction pathways that putatively participate in processing the resetting stimuli. Output pathways will be investigated, with respect to varying physiological parameters that are hypothesize to be the first- or second-stage targets of clock- gene functions. The latter includes cyclically varying levels of the encoded mRNAs and proteins; these molecular cyclings will be tracked in conjunction with physiological recordings and perturbations, by applying a transgene in which portions of a clock gene (called period) have been fused to DNA sequences encoding a real-time reporter; this is luciferase activity, which as recently been shown to permit non-invasive monitoring of molecular rhythms in live adult flies and in per-expressing tissues explanted from animals late in development. Genetic and molecular-genetic studies are proposed in two areas: (1) analysis of the behavioral and neurobiological consequences of manipulating a clock-controlled gene~s expression, and of effecting the same kinds of perturbations of a neuropeptide-encoding gene whose product is co-expressed with clock genes in a subset of the CNA pacemaking neurons; (2) genetic and biological studies of rhythm mutants, recently isolated on the basis of defects in circadian behavioral rhythms: these phenogenetic studies will proceed into cloning of the genes defined by mutations that appear to be the most promising candidates for disrupting important elements of Drosophila~s circadian system.

昼夜节律是有机无处不在的生物周期 谁的每日监管与井有密切相关 从微生物到哺乳动物的物种。 25- 一年历史的理解这些节奏的遗传方法 最近开始揭示时钟机制的元素 是每日节奏的基础。 该计划中的这个子词 项目应用围绕行为遗传，神经 - 昼夜节律的遗传学和分子神经生物学 果蝇。实验提出的压力更多是一个系统〜 方法比集中在坚硬的核心的方法 分子起搏。 因此，行为的神经底物 节奏和后期发展的定期特征将是 通过施用节奏突变体和转基因来研究 带有操纵形式的时钟创作的菌株；这些研究 包括对神经元解剖输出途径的描述 是CNS-PACEMAKER细胞的候选者，并且有选择地 影响此类细胞在 与生物鉴定分子的作用的结合 介导的神经元损伤。通往中央起搏器的输入路径， 引入环境提示，以实现至关重要的每日重新设置 时钟将根据解剖学和元素进行解剖 推定参与的信号转移途径 处理重置刺激。 输出途径将是 研究了，关于不同的生理参数 假设是时钟的第一或第二阶段目标 基因功能。 后者包括周期性变化的水平 编码的mRNA和蛋白质；这些分子循环将是 与生理记录和 扰动，通过应用一个转基因 时钟基因（称为周期）已融合到DNA序列 编码实时记者；这是荧光素酶活性，作为 最近被证明允许对 活蝇和表达组织中的分子节奏 在发育后期从动物中展开。 遗传和 在两个领域提出了分子遗传学研究：（1）分析 的行为和神经生物学后果 操纵时钟控制的基因表达和效应 神经肽编码基因的相同类型的扰动 其产品与时钟基因共表达 CNA起搏神经元； （2）遗传和生物学研究 节奏突变体，最近基于缺陷而分离 昼夜节律节奏：这些表性研究将 继续克隆由突变定义的基因 成为破坏重要的最有前途的候选人 果蝇昼夜节律系统的元素。