Genetic Analysis of Feeding Behavior and Fat Deposition

摄食行为和脂肪沉积的遗传分析

• 批准号: 7564783
• 负责人: ALAN S KOPIN
• 金额: $ 34.9万
• 依托单位: TUFTS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7564783
• 关键词: Behavioral Genetics; Biochemical; Bioinformatics; Biological Assay; Biological Models; Candidate Disease Gene; Collection; Complement; Deposition; Drosophila genus; Drosophila melanogaster; Eating; Fatty Acids; Fatty acid glycerol esters; Feeding behaviors; Functional disorder; Gene Targeting; Genes; Genetic; Genetic Models; Genetic Screening; Genetic Transcription; Goals; Histologic; Homologous Gene; Human; Intestines; Investigation; Laboratories; Link; Lipids; Mammals; Maps; Mediating; Mediator of activation protein; Metabolism; Methodology; Molecular; Obesity; Pathway interactions; Phenotype; Physiological; Principal Investigator; Proteins; RNA Interference; Role; Scanning Electron Microscopy; Screening procedure; Selection Criteria; Serotonin; Signal Transduction; Staging; Tissues; Transgenic Organisms; Tryptophan 5-monooxygenase; base; feeding; fly; genetic analysis; inhibitor/antagonist; meetings; mutant; novel; obesity treatment; receptor; receptor-mediated signaling; serotonin receptor; success; tool

Important mechanisms underlying the control of feeding and fat deposition have been conserved between Drosophila melanogaster and human. This validates the fruit fly as a genetic model system for identifying and characterizing novel genes that mediate metabolic processes. In Preliminary Studies, we have demonstrated that serotonin (5-hydroxytryptamine, 5HT) acts as a potent inhibitor of fly feeding behavior through activation of one or more 5HT receptors. These findings provide a direct parallel with mammals, for which it has been established that the 5HT2C receptor is an important mediator of feeding behavior and a well recognized target for the treatment of obesity. Aim 1 of this application proposes to further explore the molecular mechanisms underlying the serotonergic control of feeding in Drosophila, utilizing a combination of existing mutants and RNA interference flies which are currently being generated in our laboratory. These strains will provide tools to investigate the extent to which different 5HT receptor subtypes as well as their tissue specific expression (e.g. CMS vs. intestine) underlie the serotonergic control of feeding. The highly sensitive food intake assay that we have established for assessing 5HT mediated food intake has also been utilized to begin a forward genetic screen (Aim 2). To date, 1315 strains of mutant Drosophila, each carrying a single precisely mapped insertion, have been screened for abnormalities in feeding. Twelve of the most promising candidates have been selected for detailed analysis (Aim 2A). To explore the role of the , candidate gene (and its corresponding mammalian homolog) in modulating feeding and/or fat deposition, a combination of approaches will be used, including biochemical (e.g. fatty acid profiling), histologic (e.g. scanning electron microscopy of fat tissue), and genetic (e.g. transgenic rescue) methodologies. Based on our initial success in identifying candidate genes, we propose to continue the forward genetic screen (Aim 2B). The goal of this subsequent effort is to define modifiers of (i) food intake under basal conditions (ii) food intake in the presence of 5HT (to identify modifiers of feeding within the 5HT and intersecting pathways) and (iii) fat deposition. To enhance the likelihood of finding a phenotype and of identifying a gene with physiologic relevance in humans, we will utilize bioinformatic tools to pre-select insertion strains for screening. The selection criteria will include (i) disruption of the transcription unit, and (ii) existence of a mammalian homolog corresponding to the targeted fly gene. To date, -2,500 insertion bearing strains which meet these criteria have been identified. Each of these strains will be assessed for alterations in feeding behavior and fat deposition. Outliers will be characterized as described above. We anticipate that the proposed studies will identify novel genes relevant to the pathophysiology of human obesity.

控制摄食和脂肪沉积的重要机制在 果蝇和人类。这验证了果蝇作为识别遗传模型系统的有效性 并表征介导代谢过程的新基因。在初步研究中，我们有 证明血清素（5-羟色胺，5HT）是苍蝇进食行为的有效抑制剂 通过激活一种或多种5HT受体。这些发现与哺乳动物提供了直接的相似之处，因为 已经确定 5HT2C 受体是摄食行为的重要介质， 公认的治疗肥胖的目标。本申请的目标 1 建议进一步探索 果蝇血清素能控制摄食的分子机制，利用以下组合 现有的突变体和RNA干扰果蝇目前正在我们的实验室中产生。这些 菌株将提供工具来研究不同 5HT 受体亚型及其作用的程度 组织特异性表达（例如 CMS 与肠道）是进食的血清素控制的基础。高度 我们为评估 5HT 介导的食物摄入而建立的敏感食物摄入测定也已被 用于开始正向遗传筛选（目标 2）。迄今为止，已有 1315 株突变果蝇，每株都携带 一个精确映射的插入，已被筛查出喂养异常。最多的十二个 已选择有希望的候选者进行详细分析（目标 2A）。为了探索 的作用， 调节摄食和/或脂肪沉积的候选基因（及其相应的哺乳动物同源物）， 将使用多种方法的组合，包括生化（例如脂肪酸分析）、组织学（例如组织学） 脂肪组织的扫描电子显微镜）和遗传（例如转基因救援）方法。基于 我们在识别候选基因方面取得了初步成功，我们建议继续进行正向遗传筛选 （目标 2B）。随后努力的目标是定义（i）基础条件下食物摄入量的调节因素 (ii) 存在 5HT 的情况下的食物摄入量（以确定 5HT 内喂养的调节因素和相交的因素） 途径）和（iii）脂肪沉积。提高发现表型和鉴定基因的可能性 与人类的生理相关性，我们将利用生物信息学工具预先选择插入菌株 筛选。选择标准将包括（i）转录单位的破坏，以及（ii）存在 与目标果蝇基因相对应的哺乳动物同源物。迄今为止，-2,500 个插入轴承应变 已确定满足这些标准。将评估每种菌株的喂养变化 行为和脂肪沉积。异常值的特征将如上所述。我们预计 拟议的研究将鉴定与人类肥胖病理生理学相关的新基因。

项目成果

A critical role for the Drosophila dopamine D1-like receptor Dop1R2 at the onset of metamorphosis.

• DOI: 10.1186/s12861-016-0115-z
• 发表时间: 2016-05-16
• 期刊: BMC developmental biology
• 作者: Regna K;Kurshan PT;Harwood BN;Jenkins AM;Lai CQ;Muskavitch MA;Kopin AS;Draper I
• 通讯作者: Draper I