Identifying genes required for digestive physiology and lipid metabolism

描述（由申请人提供）：异常脂质代谢有助于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）。提高我们对脂质代谢及其相关疾病的理解的一个主要障碍是，在活生物体中进行的代谢研究太少了。因此，协调脂肪酸（FA）的吸收和运输以及脂肪组织中形态发生和脂肪储存的动态调节信号仍不清楚。为了解决这一差距，该研究小组开创了在活斑马鱼中由许多细胞类型组成的复杂器官中成像脂质摄取，运输和储存的方法。法伯实验室已经建立了工具来可视化斑马鱼幼虫中膳食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。这些最先进的体内光学报告提供了器官生理学的全面观点，而不是在以前的器官发育研究中揭示的。在Derek Stemple博士的指导下，我们建议使用这些方法筛选斑马鱼突变资源产生的突变系。我们将首先通过饲喂荧光脂类和亲脂染料，并分析其在活幼虫中的积累模式，对消化器官脂质摄取、代谢、运输和储存方面的缺陷进行初步筛选，以确定突变体。然后，我们将进行二次筛选，以全面表征已确定的脂质代谢和脂肪组织突变体的表型。拟议研究的总体目标是确定斑马鱼脂质摄取，运输和储存的重要遗传修饰因子。其基本原理是，一旦知道了调节斑马鱼脂质代谢的遗传途径，这些信息就可以转化为人类，通过控制选定组织中不同的脂质代谢过程来启动新的治疗方法，以降低心血管疾病、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.