Aquaporins in Drosophila melanogaster

果蝇中的水通道蛋白

• 批准号: 6805227
• 负责人: NANCY KAUFMANN
• 金额: $ 4.89万
• 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/6805227
• 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果蝇中水通道通道的功能。该赠款申请的具体目的是：1）使用MRNA原位和免疫组织化学来揭示假定的水通道蛋白的作用部位。 2）确定假定果蝇水通道蛋白的底物特异性和孔渗透性。这将通过使用酵母囊泡表达系统来实现，并在布罗德斯基和Zeidel实验室中停止流量荧光法，并在必要时使用更传统的Xenopus laevis swilling-Swilling测定法。为了进一步将功能与位置相关联，将来，将为表现出生理相关的水的渗透性和定位而产生功能等位基因的损失。这些苍蝇中的马尔皮亚小管将在已建立的水排泄测定中进行测试。 这项工作将进一步了解水通道的结构/功能关系，以及我们对维持渗透稳态的水通道蛋白功能的理解。这项工作不仅与哺乳动物肾功能和糖尿病的研究有关，而且还将有助于我们了解昆虫水排泄，这是控制蚊子种群的主要目标和疟疾的传播。