title: Methods to identify and analyze gene products involved in neuronal intracellular transport using Drosophila.
abstract: Proper neuronal function critically depends on efficient intracellular transport and disruption of transport leads to neurodegeneration. Molecular pathways that support or regulate neuronal transport are not fully understood. A greater understanding of these pathways will help reveal the pathological mechanisms underlying disease. Drosophila melanogaster is the premier model system for performing large-scale genetic functional screens. Here we describe methods to carry out primary and secondary genetic screens in Drosophila aimed at identifying novel gene products and pathways that impact neuronal intracellular transport. These screens are performed using whole animal or live cell imaging of intact neural tissue to ensure integrity of neurons and their cellular environment. The primary screen is used to identify gross defects in neuronal function indicative of a disruption in microtubule-based transport. The secondary screens, conducted in both motoneurons and dendritic arborization neurons, will confirm the function of candidate gene products in intracellular transport. Together, the methodologies described here will support labs interested in identifying and characterizing gene products that alter intracellular transport in Drosophila.
authors.affiliation: Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, MN, USA.
authors.forename: Amanda L
authors.initials: AL
authors.lastname: Neisch
authors.affiliation: Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, MN, USA.
authors.forename: Adam W
authors.initials: AW
authors.lastname: Avery
authors.affiliation: Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: James B
authors.initials: JB
authors.lastname: Machamer
authors.affiliation: Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, MN, USA.
authors.forename: Min-gang
authors.initials: MG
authors.lastname: Li
authors.affiliation: Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, MN, USA.
authors.forename: Thomas S
authors.initials: TS
authors.lastname: Hays
mesh_terms: D000818:Animals; D001370:Axonal Transport / Q000235:genetics* / Q000502:physiology; D001369:Axons / Q000378:metabolism*; D003712:Dendrites / Q000502:physiology; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism*; D000072159:Dynactin Complex; D004398:Dyneins / Q000235:genetics / Q000378:metabolism*; D016547:Kinesins / Q000378:metabolism*; D007814:Larva / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000235:genetics / Q000378:metabolism; D019636:Neurodegenerative Diseases / Q000473:pathology; D009473:Neuronal Plasticity / Q000235:genetics / Q000502:physiology; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics
chemical_list: D029721:Drosophila Proteins; D000072159:Dynactin Complex; D008869:Microtubule-Associated Proteins; D034741:RNA, Small Interfering; D004398:Dyneins; D016547:Kinesins
keywords: Dendritic arborization neurons; Drosophila; Dynein; Genetic screens; Intracellular transport; Live cell imaging; Motoneurons; Neurodegeneration; RNAi; Tail flip