Acquisition of an Olympus SZX7 fluorescent stereo microscope for dissecting late-stage Drosophila embryos and selecting Drosophila embryos with GFP/RFP tagged genes

获取奥林巴斯 SZX7 荧光体视显微镜，用于解剖晚期果蝇胚胎并选择带有 GFP/RFP 标记基因的果蝇胚胎

• 批准号: 10795289
• 负责人: Lan Jiang
• 金额: $ 2.35万
• 依托单位: OAKLAND UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10795289
• 关键词: Air; Antibodies; Apical; Autophagosome; Award; Binding; Binding Proteins; Biological Assay; Biological Models; Blood Vessels; Cessation of life; Chromosomes; Co-Immunoprecipitations; Cytoplasm; Cytoplasmic Vesicles; Defect; Deposition; Disease; Dissection; Drosophila eye; Drosophila genus; Drug Metabolic Detoxication; Embryo; Endosomes; Epithelial Cells; Epithelium; Extracellular Matrix; Fluorescence; Gases; Gene Family; Genes; Genetic; Glycocalyx; Glycolysis; Goals; Golgi Apparatus; Human; Image; Immunohistochemistry; Individual; Insecta; Invertebrates; Kidney; Knowledge; Lactoylglutathione Lyase; Life; Light; Liquid substance; Lung; Lysosomes; Mediating; Metabolic; Microscope; Morphogenesis; Organ; Organism; Pathway interactions; Phenotype; Polycystic Kidney Diseases; Property; Proteins; Pyruvaldehyde; Recombinants; Research Project Grants; Role; Shapes; Structure; System; Testing; Time; Trachea; Tube; Tubular formation; United States National Institutes of Health; Vertebrates; Vesicle; Yeasts; body system; exosome; experimental study; fluorescence microscope; fly; gene function; in vivo; malformation; member; monolayer; mutant; protein biomarkers; protein transport; repaired; secretion process; spine bone structure; trafficking

Acquisition of an Olympus SZX7 fluorescent stereo microscope for dissecting late-stage Drosophila embryos and selecting Drosophila embryos with GFP/RFP tagged genes This proposal is for the acquisition of an Olympus SZX7 fluorescent stereo microscope to replace an old and broken fluorescence stereo microscope, which was used to select GFP- positive fly embryos and dissect fly embryos when ring light is provided. 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. 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 lumen. Then, this lumen is cleared before air fills the lumen. The goal 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 the trafficking of the apical luminal matrix during tube maturation. Aim. 2: Determine the function of Osi genes as coordinators to control endosome-mediated protein trafficking. Aim. 3: Identify proteins that directly bind to Osi proteins. To carry out the proposed experiments, we will analyze protein trafficking and structural changes in late-stage embryos by immunostaining. In addition, we will analyze the dynamic coordination of vesicular trafficking in vivo by observing GFP-tagged vesicle markers and RFP-tagged protein markers simultaneously in Osi mutant background. The tracheal phenotypes in Osi mutants appear in late-stage embryos (mid-stage 17). Due to cuticle formation, antibodies cannot enter embryos effectively without dissection. Therefore, we need to dissect the late-stage embryos before immunostaining. In addition, we will generate fly strains that carry GFP-tagged vesicle markers and RFP-tagged luminal protein markers in Osi mutant background. Since some marker genes are located on the same chromosome as Osi genes, we will recombine these marker genes to Osi mutant background genetically. Therefore the potential live recombinant fly embryos that carry GFP- or RFP- tagged marker genes can be selected using a stereo fluorescent microscope. Unfortunately, our old fluorescence stereo microscope that can be used to dissect late-stage fly embryos as well as to select live embryos that carry fluorescently tagged genes is broken beyond repair. Thus, this requested Olympus SZX7 fluorescent stereo microscope will allow us to carry out the proposed experiments (Aim. 1 and Aim. 2) on time. Without it, our progress on the project will be significantly delayed.

获得奥林巴斯SZX7荧光立体显微镜用于晚期解剖

项目成果

The novel gene apnoia regulates Drosophila tracheal tube size.

新基因 apnoia 调节果蝇气管的大小。

• DOI: 10.1002/dvdy.29
• 发表时间: 2019
• 期刊: Developmental dynamics : an official publication of the American Association of Anatomists
• 作者: Scholl,Aaron;O'Brien,MichaelJ;Chandran,RachanaR;Jiang,Lan
• 通讯作者: Jiang,Lan

The Osiris family genes function as novel regulators of the tube maturation process in the Drosophila trachea.

• DOI: 10.1371/journal.pgen.1010571
• 发表时间: 2023-01
• 期刊: PLoS genetics
• 影响因子: 4.5