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以下两种情况下是保守的： 果蝇和人类。这验证了果蝇作为一个遗传模型系统， 以及表征介导代谢过程的新基因。在初步研究中，我们 5-羟色胺（5-hydroxytryptamine，5 HT）对果蝇的摄食行为有抑制作用 通过激活一种或多种5 HT受体。这些发现提供了与哺乳动物的直接平行，因为 已经确定5 HT 2C受体是摄食行为的重要介质， 公认的治疗肥胖症的靶点。本申请的目标1提出进一步探索 果蝇摄食的雌激素能控制的分子机制，利用 现有的突变体和RNA干扰苍蝇，目前正在产生在我们的实验室。这些 菌株将提供研究不同5 HT受体亚型及其 组织特异性表达（例如CMS相对于肠）是摄食的多巴胺能控制的基础。高度 我们建立的用于评估5-HT介导的食物摄入的敏感食物摄入测定也被 用于开始正向遗传筛选（Aim 2）。到目前为止，1315株突变果蝇，每一株都携带 一个精确定位的插入，已经被筛选出进食异常。十二个最 已选定有希望的候选者进行详细分析（目标2A）。为了探讨的作用， 在调节进食和/或脂肪沉积中的候选基因（及其相应的哺乳动物同源物）， 将使用多种方法的组合，包括生物化学（例如脂肪酸谱），组织学（例如， 脂肪组织的扫描电子显微镜）和遗传（例如转基因拯救）方法学。基于 我们在确定候选基因方面取得了初步成功，我们建议继续进行遗传筛查。 (Aim 2B）。这一后续努力的目标是定义修饰符（i）在基础条件下的食物摄入量 (ii)在5 HT存在下的食物摄入量（以确定5 HT内的摄食调节剂和交叉调节剂） 途径）和（iii）脂肪沉积。为了提高发现表型和识别基因的可能性， 与人类生理相关，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，和（ii）转录单位的存在。 对应于靶向的果蝇基因的哺乳动物同源物。到目前为止，-2，500个插入轴承菌株， 符合这些标准的已经确定。这些菌株中的每一个都将被评估喂养的改变 行为和脂肪沉积。离群值将如上所述进行表征。我们预计 提出的研究将鉴定与人类肥胖症的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