title: Using membrane-targeted green fluorescent protein to monitor neurotoxic protein-dependent degeneration of Drosophila eyes.
abstract: Age-related neurodegeneration has been studied extensively through the use of model organisms, including the genetically versatile Drosophila melanogaster. Various neurotoxic proteins have been expressed in fly eyes to approximate degeneration occurring in humans, and much has been learned from this heterologous system. Although Drosophila expedites scientific research through rapid generational times and relative inexpensiveness, one factor that can hinder analyses is the examination of milder forms of degeneration caused by some toxic proteins in fly eyes. Whereas several disease proteins cause massive degeneration that is easily observed by examining the external structure of the fly eye, others cause mild degeneration that is difficult to observe externally and requires laborious histological preparation to assess and monitor. Here, we describe a sensitive fluorescence-based method to observe, monitor, and quantify mild Drosophila eye degeneration caused by various proteins, including the polyglutamine disease proteins ataxin-3 (spinocerebellar ataxia type 3) and huntingtin (Huntington's disease), mutant α-synuclein (Parkinson's disease), and Aβ42 (Alzheimer's disease). We show that membrane-targeted green fluorescent protein reports degeneration robustly and quantitatively. This simple yet powerful technique, which is amenable to large-scale screens, can help accelerate studies to understand age-related degeneration and to find factors that suppress it for therapeutic purposes.
authors.affiliation: Graduate Program in Cancer Biology, Wayne State University School of Medicine, Detroit, Michigan; Department of Pharmacology, Wayne State University School of Medicine, Detroit, Michigan.
authors.forename: Aaron A
authors.initials: AA
authors.lastname: Burr
authors.forename: Wei-Ling
authors.initials: WL
authors.lastname: Tsou
authors.forename: Gorica
authors.initials: G
authors.lastname: Ristic
authors.forename: Sokol V
authors.initials: SV
authors.lastname: Todi
mesh_terms: D016229:Amyloid beta-Peptides / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D000067699:Ataxin-3; D016827:CD8 Antigens / Q000235:genetics / Q000378:metabolism; D004195:Disease Models, Animal; D004330:Drosophila; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D000071058:Huntingtin Protein; D008822:Mice, Transgenic; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D012162:Retinal Degeneration / Q000175:diagnosis* / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D051844:alpha-Synuclein / Q000235:genetics
chemical_list: D016229:Amyloid beta-Peptides; D016827:CD8 Antigens; D029721:Drosophila Proteins; C557617:GAL4 protein, Drosophila; C542005:Htt protein, Drosophila; D000071058:Huntingtin Protein; D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D010446:Peptide Fragments; D010455:Peptides; D012097:Repressor Proteins; D014157:Transcription Factors; D051844:alpha-Synuclein; C075222:amyloid beta-protein (1-42); D049452:Green Fluorescent Proteins; C097188:polyglutamine; C092341:ATXN3 protein, human; D000067699:Ataxin-3
keywords: Alzheimer's; Huntington's; Parkinson's; neurodegeneration; polyglutamine; spinocerebellar ataxia