Aquaporins in Drosophila melanogaster

果蝇中的水通道蛋白

• 批准号: 6930554
• 负责人: NANCY KAUFMANN
• 金额: $ 5.15万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-29 至 2006-09-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6930554
• 关键词: Drosophilidae; Xenopus oocyte; arthropod genetics; body water; fluorimetry; gene expression; homeostasis; immunocytochemistry; membrane permeability; postdoctoral investigator; protein localization; protein structure function; water channel; yeasts

DESCRIPTION (provided by applicant): The long-term goal of this project is to elucidate the function of aquaporin water channels in Drosophila melanogaster. The specific aims of this grant application are to: 1) Use mRNA in situ and immunohistochemistry to uncover the sites of action of putative aquaporins. 2) Determine the substrate specificity and pore permeability of putative Drosophila aquaporins. This will be accomplished by using the yeast vesicle expression system and stopped-flow fluorometry pioneered in the Brodsky and Zeidel laboratories and, if necessary, the more traditional Xenopus laevis egg-swelling assay. To further correlate function with location, in the future, loss of function alleles will be generated for candidates showing physiologically relevant water permeability and localization. Malpighian tubules from these flies will be tested in established water excretion assays. This work will further our understanding of the structure/function relationship for water channels, as well as our understanding of aquaporin function in maintaining osmotic homeostasis. This work is relevant not only to studies of mammalian renal function and Diabetes insipidus, but it will also help us understand insect water excretion, a prime target for controlling mosquito populations and the spread of malaria.

描述（由申请人提供）： 该项目的长期目标是阐明果蝇水通道蛋白水通道的功能。本次资助申请的具体目标是：1) 使用原位 mRNA 和免疫组织化学来揭示假定的水通道蛋白的作用位点。 2) 确定假定的果蝇水通道蛋白的底物特异性和孔渗透性。这将通过使用布罗德斯基和泽德尔实验室首创的酵母囊泡表达系统和停流荧光测定法来实现，如有必要，还可以使用更传统的非洲爪蟾卵膨胀测定法。为了进一步将功能与位置关联起来，将来，将为表现出生理相关的水渗透性和定位的候选者生成功能丧失等位基因。这些果蝇的马氏小管将在已建立的水排泄测定中进行测试。 这项工作将进一步加深我们对水通道的结构/功能关系的理解，以及我们对水通道蛋白在维持渗透压稳态中的功能的理解。这项工作不仅与哺乳动物肾功能和尿崩症的研究相关，而且还将帮助我们了解昆虫的水排泄，这是控制蚊子种群和疟疾传播的主要目标。