Identifying genes required for digestive physiology and lipid metabolism

描述(由申请人提供)：异常的脂代谢导致多种人类疾病的病因，包括心血管疾病(CVD)、肥胖和胰岛素抵抗(IR)。提高我们对脂代谢及其相关疾病的了解的一个主要障碍是在活体生物中进行的代谢研究如此之少。因此，协调脂肪组织中脂肪酸(FA)的吸收和运输、形态发生和脂肪储存的动态调控信号仍然不清楚。为了解决这一差距，聚集在一起的研究团队开创了在活斑马鱼的多种细胞类型组成的复杂器官中对脂肪的吸收、运输和储存进行成像的方法。Farber实验室已经建立了工具来可视化斑马鱼幼体中饲料FA的细胞动力学，而罗尔斯实验室已经开发了补充方法，使用重要的荧光亲脂染料来可视化斑马鱼脂肪组织。这些最先进的活体光学记者提供了器官生理学的全面视角，这是以前对器官发育的研究中没有揭示的。我们建议使用这些方法在Derek Stemple博士的指导下筛选由斑马鱼突变资源产生的突变系。我们将首先进行初步筛选，通过饲养荧光脂类和亲脂染料并分析它们在活体幼虫中的积累模式，来鉴定在消化器官脂肪吸收、代谢、运输和储存方面存在缺陷的突变体。然后，我们将进行二次筛选，以全面表征已鉴定的脂代谢和脂肪组织突变的表型。这项拟议研究的总体目标是确定斑马鱼中脂类吸收、运输和储存的重要遗传修饰物。其基本原理是，一旦知道了调控斑马鱼脂肪代谢的遗传途径，这些信息就可以转化到人类身上，启动新的治疗方法，通过控制选定组织中不同的脂肪代谢过程来降低心血管疾病、肥胖、IR和相关疾病的风险。

DESCRIPTION (provided by applicant): Aberrant lipid metabolism contributes to the etiology of multiple human diseases including cardiovascular disease (CVD), obesity, and insulin resistance (IR). A major impediment to improving our understanding of lipid metabolism and its related disorders is that so few metabolic studies have been carried out in live organisms. As a result, the dynamic regulatory signals that coordinate absorption and transport of fatty acids (FA), and morphogenesis and fat storage in adipose tissues, remain unclear. To address this gap, the assembled research team has pioneered methods to image lipid uptake, transport and storage, within complex organs composed of many cell types in live zebrafish. The Farber lab has established tools to visualize the cellular dynamics of dietary FA in zebrafish larvae, while the Rawls lab has developed complementary methods for using vital fluorescent lipophilic dyes to visualize zebrafish adipose tissues. These state-of-the-art in vivo optical reporters provide a comprehensive view of organ physiology not revealed in previous studies of just organ development. We propose to use these methods to screen mutant lines generated by the Zebrafish Mutation Resource under the direction of Dr. Derek Stemple. We will first conduct a primary screen to identify mutants defective in digestive organ lipid uptake, metabolism, transport and storage by feeding fluorescent lipids and lipophilic dyes and assaying their patterns of accumulation in live larvae. We will then conduct secondary screens to comprehensively characterize the phenotypes of identified lipid metabolism and adipose tissue mutants. The overall objective of the proposed research is to identify important genetic modifiers of lipid uptake, transport, and storage in the zebrafish. The rationale is that, once the genetic pathways regulating zebrafish lipid metabolism are known, this information could be translated to humans to initiate new therapeutic approaches to reduce risk of CVD, obesity, IR, and associated disorders by controlling distinct lipid metabolic processes in selected tissues.