CIRCADIAN SYSTEM AND PRENATAL COCAINE IN ZEBRAFISH

斑马鱼的昼夜节律系统和产前可卡因

• 批准号: 8245613
• 负责人: IRINA V. ZHDANOVA
• 金额: $ 31.21万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8245613
• 关键词: Address; Adult; Affect; Age; Animals; Area; Attenuated; Behavioral; Biological; Blood Vessels; Brain; Bromodeoxyuridine; Cell Cycle; Cell Proliferation; Cell division; Circadian Rhythms; Cocaine; Cocaine Dependence; Complementary DNA; Confocal Microscopy; Control Groups; Cyclin A; Cyclin-Dependent Kinases; Data; Development; Dopamine; Dose; Embryo; Embryonic Development; Evaluation; Fertilization; Fetal Cocaine Exposure; Fishes; Forebrain Development; Funding; Future; Gender; Gene Expression; Genes; Genetic; Genetic Transcription; Genetic Variation; Goals; Growth; Health; High Pressure Liquid Chromatography; Hormonal; Hormones; Individual; Individual Differences; Lead; Light; Limbic System; Link; Maintenance; Mammals; Melatonin; Melatonin Receptors; Messenger RNA; Methods; Molecular; Morphology; Motor Activity; Nature; Neurons; Outcome; Pattern; Pharmaceutical Preparations; Phase; Physiologic pulse; Physiological; Physiological Processes; Preventive; Process; Regulation; Research; Resources; Reverse Transcriptase Polymerase Chain Reaction; Rewards; Role; S Phase; Sampling; Signal Transduction; Sleep; Somatotropin; Staging; Staining method; Stains; Structure; System; Systems Development; Techniques; Telencephalon; Testing; Therapeutic; Time; Transgenic Organisms; Tubulin; Up-Regulation; Vascular blood supply; Withdrawal; Work; Zebrafish; aged; base; brain size; brain volume; circadian pacemaker; cocaine exposure; combat; critical period; design; dopamine transporter; genetic analysis; genetic strain; improved; insight; neurogenesis; neuron development; neuronal patterning; novel therapeutic intervention; prenatal; prevent; prophylactic; public health relevance; response; sample collection

DESCRIPTION (provided by applicant): The goal of the proposed project is to establish the role of the circadian system in the prenatal effects of cocaine exposure during a critical period of embryonic brain development in a diurnal vertebrate, zebrafish. These studies will be based on the results of the previous 3-year funding period, which established a reciprocal relationship between the cocaine and zebrafish clock system, found that melatonin can counteract the effects of cocaine on brain development, identified two strains with contrasting effects of cocaine on circadian system and behavioral effects of cocaine, and characterized the behavioral and molecular effects of cocaine and cocaine withdrawal in adult and aged zebrafish of both genders. The continuation of this research will specifically focus on characterization of the effects of cocaine and circadian clock system on brain development, and address the hypothesis that reciprocal interaction between the two is due to their contrasting effects on neuronal proliferation during early stages of brain formation. The Specific Aims will address: 1) In-depth characterization of the effects of early cocaine and melatonin exposure on zebrafish brain development and gene expression; 2) An impact of the earlier identified genetic variation in circadian response to cocaine on the effects of prenatal cocaine and melatonin exposure; 3) The role of circadian deficits in the prenatal brain development and effects of cocaine, and whether exogenous melatonin can counteract these effects. Zebrafish will be exposed to cocaine during the major periods of cortical (pallium) and limbic system (dorsomedial and dorsolateral telencephalon) development, synchronized with either the nighttime or daytime circadian phase. Melatonin will be administered prior to cocaine at different circadian phases, in embryos with normal and deficient circadian system. The brain size, cell division rate, dopamine levels, expression of the circadian genes and those encoding for the melatonin receptors, dopamine transporter, cyclin-dependent kinases, cyclin A and growth hormone will be evaluated throughout development and maturation. The time of development and integrity of the neuronal ensembles will be evaluated in transgenic fish with fluorescent markers for alpha-1-tubulin and f-spondin. The cocaine-induced changes in the locomotor activity and sleep will be documented during early post-natal period. Understanding the role of the circadian factors in brain development and mechanisms through which they attenuate prenatal effects of cocaine, should help to design preventive and therapeutic strategies to counteract prenatal cocaine impact on early brain development or subsequent changes in brain functions. PUBLIC HEALTH RELEVANCE: The biological circadian clock system can modulate the effects of cocaine. This project will address the role of the circadian factors, including melatonin, in the effects of prenatal cocaine exposure on brain development and molecular processes, using a diurnal, genetically well-characterized vertebrate. The results of this study should provide understanding of the mechanisms involved in reciprocal interaction between the prenatal effects of cocaine and circadian system, and lead to the development of new prophylactic or therapeutic strategies.

描述（由申请人提供）：拟议项目的目的是确定昼夜节律系统在可卡因暴露的产前效应中的作用，这是Zebrafish昼夜脊椎动物中胚胎脑发育的关键时期。这些研究将基于上一个三年资金期间的结果，该期间建立了可卡因和斑马鱼时钟系统之间的相互关系，发现褪黑激素可以抵消可卡因对脑发育的影响，确定了两种菌株，与可卡因对昼夜节律和cocaine and Cocaine和特征性行为效果的对比造成了鲜明的影响，以及对CACAINAL和特征性行为影响CACEARCAINAL和CACAINTAL效果的影响。两种性别的老年斑马鱼。这项研究的延续将特别关注可卡因和昼夜节律系统对脑发育的影响的表征，并解决了以下假设：两者之间的相互相互作用是由于它们在大脑形成的早期阶段对神经元增殖的影响。具体目的将解决：1）早期可卡因和褪黑激素暴露对斑马鱼脑发育和基因表达的影响； 2）早期发现可卡因反应中遗传变异对产前可卡因和褪黑激素暴露的影响的影响； 3）昼夜节律缺陷在可卡因的产前脑发育和作用中的作用，以及外源褪黑激素是否可以抵消这些作用。斑马鱼将在主要时期（pallium）和边缘系统（背侧和背外侧端脑）发育的主要时期暴露于可卡因，并与夜间或白天昼夜节律同步。褪黑激素将在可卡因之前在不同的昼夜节相中，在具有正常且缺乏昼夜节律系统的胚胎中给药。在整个发育和成熟过程中，将评估脑大小，细胞分裂速率，多巴胺水平，昼夜节律基因的表达以及编码褪黑激素受体，多巴胺转运蛋白，细胞周期蛋白依赖性激酶，细胞周期蛋白A和生长激素的基因的表达。神经元集合的发育和完整性的时间将在具有α-1-微管蛋白和F型蛋白的荧光标记的转基因鱼类中进行评估。可卡因诱导的运动活动和睡眠的变化将在产后早期记录。了解昼夜节律因素在大脑发育和减轻可卡因产前作用的机制中的作用，应有助于设计预防性和治疗性策略，以抵消产前可卡因对早期大脑发育或随后大脑功能的变化。公共卫生相关性：生物昼夜节律系统可以调节可卡因的影响。该项目将使用昼夜，遗传良好的脊椎动物来解决昼夜节律因素（包括褪黑激素）对脑发育和分子过程的作用。这项研究的结果应提供对可卡因和昼夜节律系统产前效应之间相互作用涉及的机制的理解，并导致新的预防性或治疗策略的发展。