Transcriptional output pathways of the circadian clock

生物钟的转录输出途径

基本信息

批准号：
7359643
负责人：
HERMAN WIJNEN
金额：
$ 22.08万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-03-01 至 2012-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7359643
关键词：
Adult Affect Animal Model Animals Behavior Behavioral Binding Biological Assay Body Temperature Brain Cardiovascular Physiology Cell Line Cells Chimeric Proteins Circadian Rhythms Condition Daily Diabetes Mellitus Diagnostic Disease Disruption Drosophila genus Drosophila melanogaster Engineering Ensure Feedback Gene Mutation Genes Genetic Transcription Glucocorticoid Receptor Hormonal Human Human body Insecta Luciferases Malignant Neoplasms Mammals Metabolism Molecular Mutation Neurons Output Pacemakers Pathway interactions Phenotype Physiology Procedures Protein Biosynthesis RNA Regulation Reporter Respiration Schedule Sleep Disorders Sleep Wake Cycle Testing Time Transcript Transcriptional Regulation Transgenic Organisms Variant base chromatin immunoprecipitation circadian pacemaker design fly in vivo inhibitor/antagonist insight knock-down promoter research study response transcription factor

项目摘要

DESCRIPTION (provided by applicant): Circadian clocks in the bodies of humans ensure that functions such as the sleep/wake cycle, body temperature, hormonal secretion, cardiovascular function, respiration and metabolism are timed correctly. There is an exceptionally high degree of conservation between the circadian clocks of mammals and insects and much of our current understanding of clock function in humans is based on studies of the genes and the molecular circuits of the clock in model organisms such as Drosophila. Transcriptional feedback circuits responsible for the autonomous pacemaker function of the Drosophila clock have been identified in detail, but much less is known about the pathways connecting these circuits to overt rhythms in physiology and behavior. This proposal is designed to test the hypothesis that the CLK/CYC transcription factor of the Drosophila clock circuits has direct transcriptional targets in the Drosophila brain that generate behavioral rhythms. Specific Aims: (1) Transcriptional targets specific to the circadian transcription factor CLK/CYC will be identified with the use of a set of transgenic flies that allow expression of each of these transcription factors to be induced individually in clock-bearing cells. (2) Direct transcriptional targets for CLK/CYC will be identified by luciferase reporter assays and chromatin immunoprecipitation experiments performed in Drosophila S2 cells. (3) Locomotor and eclosion behavior phenotypes associated with primary transcriptional clock output in the brain will be determined with the use of transgenic flies. Significance: The insight provided by the proposed studies into the internal time keeping mechanisms of animals is not only relevant to diseases associated with disruptions of these mechanisms (sleep disorders, cancer, diabetes), but also more generally to diagnostic and treatment procedures involving bodily functions that show regular daily variations.

描述（由申请人提供）：人体的生物钟确保睡眠/觉醒周期、体温、荷尔蒙分泌、心血管功能、呼吸和新陈代谢等功能的计时正确。哺乳动物和昆虫的生物钟之间存在着极高程度的保守性，我们目前对人类生物钟功能的理解大部分是基于对果蝇等模式生物的基因和生物钟分子回路的研究。负责果蝇时钟自主起搏器功能的转录反馈回路已被详细识别，但对于将这些回路与生理和行为中的明显节律连接起来的途径知之甚少。该提案旨在测试果蝇时钟电路的 CLK/CYC 转录因子在果蝇大脑中具有产生行为节律的直接转录目标的假设。具体目标：（1）将使用一组转基因果蝇来鉴定昼夜节律转录因子 CLK/CYC 特异的转录靶标，这些转基因果蝇允许在时钟承载细胞中单独诱导这些转录因子中的每一个的表达。 (2) CLK/CYC 的直接转录靶标将通过在果蝇 S2 细胞中进行的荧光素酶报告基因测定和染色质免疫沉淀实验来鉴定。 (3)将使用转基因果蝇来确定与大脑中初级转录时钟输出相关的运动和羽化行为表型。意义：拟议的研究对动物内部计时机制提供的见解不仅与这些机制破坏相关的疾病（睡眠障碍、癌症、糖尿病）有关，而且更广泛地与涉及身体功能的诊断和治疗程序有关显示出每日有规律的变化。