Acquisition of a Drosophila chamber for culturing Drosophila strains

这项提议是为了购买一个果蝇小屋，以取代一个陈旧破旧的 在本实验室用来培养果蝇品系。获奖的NIH R15 研究项目是研究一个知之甚少的Osiris(OSI)基因家族在调节 果蝇气管中的管子成熟是揭示基本原理的首要系统 管状器官形成的机制。输卵管畸形会导致各种人类疾病， 如多囊肾病和血管疾病。 果蝇的气管是由上皮管和单层上皮层组成的分支网络。 根尖管腔周围的细胞。在输卵管成熟过程中，顶端分泌物破裂沉积。 大量的管腔基质成分进入根尖细胞外腔。然后，这个流明 在空气填满管腔之前就被清除了。先前的研究表明，蛋白质的成分 运输途径与腔蛋白的分泌/清除有关。这个 获奖项目的目的是揭示Osi蛋白作为“更广泛的协调者”的功能。 在输卵管成熟过程中的蛋白质运输。为了验证这一假设，我们将完成以下工作 三个具体目标：目标。1确定Osi基因在根尖管腔运输中的功能 在试管成熟过程中的基质。瞄准。2：确定OSI基因作为协调者的功能 控制人口贩运组成部分。瞄准。3：鉴定与Osi蛋白直接结合的蛋白质。履行 这些目标，我们将产生携带内体标记、腔蛋白标记的苍蝇品系 在Osi突变背景中检测Osi患者内小体和腔蛋白的变化 变种人。然后，我们将建立在Osi中表达Osi转基因或内体转基因的品系 突变株来分析救援效果。此后，我们将产生携带个体的苍蝇品系 Osi突变体的内体突变检测Osi基因与内体的遗传相互作用 基因。最后，我们将在气管中表达HA标记的OSI基因，以分离相互作用的蛋白质 与Osi蛋白质结合。 因此，为了进行拟议的研究，携带多种突变的苍蝇菌株， 平衡者，转基因已经产生，并将产生。目前，苍蝇品系正在培养 在实验室的室温下。不幸的是，为该项目产生了数十个菌株 由于气温的意外大幅下降或上升而丢失。建议数 日内瓦科学公司的DR-36VL型飞室仪器维修正确 湿度、光照周期和稳定的温度将使苍蝇不仅存活下来，而且还能茁壮成长。 没有它，就不可能及时开展拟议的实验。

This proposal is for the acquisition of a Drosophila chamber to replace an old and broken chamber, which was used to culture Drosophila strains in our laboratory. The awarded NIH R15 research project is to study the role of a poorly understood Osiris (Osi) gene family in regulating tube maturation in Drosophila trachea, which is a premier system to reveal fundamental mechanisms of tubular organ formation. Malformation of tubes leads to various human diseases, such as polycystic kidney disease and vascular diseases. The Drosophila trachea is a ramifying network of epithelial tubes with a monolayer of epithelial cells surrounding an apical lumen. During tube maturation, the apical secretion burst deposits large amounts of luminal matrix components to the apical extracellular lumen. Then, this lumen is cleared before air can fill the lumen. Previous studies have shown that components of protein trafficking pathways have been implicated in the secretion/clearance of luminal proteins. The objective of the awarded project is to reveal the functions of Osi proteins as “broader coordinators” of protein trafficking during tube maturation. To test this hypothesis, we will complete the following three specific aims: Aim. 1 Determine the function of Osi genes in trafficking of apical luminal matrix during tube maturation. Aim. 2: Determine the function of Osi genes as coordinators to control trafficking components. Aim. 3: Identify proteins that directly bind to Osi proteins. To fulfill these goals, we will generate fly strains that carrying endosome markers, luminal protein markers in Osi mutant background to determine changes of endosomes and luminal proteins in Osi mutants. Then, we will generate lines to express Osi transgenes or endosome transgenes in Osi mutants to analyze the rescue effect. Thereafter, we will generate fly strains carrying individual endosome mutations in Osi mutants to test genetic interaction between Osi genes and endosome genes. Finally, we will express HA-tagged Osi genes in trachea to isolate proteins that interact with Osi proteins. Therefore, to carry out the proposed research, fly strains carrying multiple mutations, balancers, and transgenes have been and will be generated. Currently the fly strains are cultured at room temperature in the lab. Unfortunately dozens of strains that were generated for the project were lost due to unexpected significant decrease or increase of the temperatures. The proposed instrument, a fly chamber, Model DR-36VL, from GENEVA SCIENTIFIC maintains proper humidity, light cycles, and stable temperatures will keep the fly strains not only alive but thriving. Without it, it will not be possible to carry out the proposed experiments in a timely manner.