title: A Behavioral Assay for Mechanosensation of MARCM-based Clones in Drosophila melanogaster.
abstract: Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense organs provide an excellent model system for the study of mechanosensation. This protocol describes a simple touch behavior in fruit flies which can be used to identify mutations that interfere with mechanosensation. The tactile stimulation of a macrochaete bristle on the thorax of flies elicits a grooming reflex from either the first or third leg. Mutations that interfere with mechanotransduction (such as NOMPC), or with other aspects of the reflex arc, can inhibit the grooming response. A traditional screen of adult behaviors would have missed mutants that have essential roles during development. Instead, this protocol combines the touch screen with mosaic analysis with a repressible cell marker (MARCM) to allow for only limited regions of homozygous mutant cells to be generated and marked by the expression of green fluorescent protein (GFP). By testing MARCM clones for abnormal behavioral responses, it is possible to screen a collection of lethal p-element mutations to search for new genes involved in mechanosensation that would have been missed by more traditional methods.
authors.affiliation: Department of Biology, College of the Holy Cross; School of Medicine, Georgetown University.
authors.forename: Timothy P
authors.initials: TP
authors.lastname: Murphy
authors.affiliation: Department of Biology, College of the Holy Cross.
authors.forename: Dan D
authors.initials: DD
authors.lastname: Luu
authors.affiliation: Department of Biology, College of the Holy Cross; Department of Biochemistry, Giesel School of Medicine, Dartmouth College.
authors.forename: Jessica A
authors.initials: JA
authors.lastname: DeSimone
authors.affiliation: Department of Biology, College of the Holy Cross; School of Medicine, Tufts University.
authors.forename: Thomas C
authors.initials: TC
authors.lastname: O'Brien
authors.affiliation: Department of Biology, College of the Holy Cross; Transgenomic Inc.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Lally
authors.affiliation: Department of Biology, College of the Holy Cross; Department of Molecular, Cell and Cancer Biology, UMass Medical School.
authors.forename: Jillian J
authors.initials: JJ
authors.lastname: Lindblad
authors.affiliation: Department of Biology, College of the Holy Cross; swebster@holycross.edu.
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Webster
mesh_terms: D000818:Animals; D001522:Behavior, Animal / Q000502:physiology*; D002999:Clone Cells; D003584:Cytological Techniques / Q000379:methods; D029721:Drosophila Proteins / Q000235:genetics; D004331:Drosophila melanogaster / Q000235:genetics / Q000502:physiology*; D004772:Entomology / Q000379:methods; D005260:Female; D008297:Male; D040542:Mechanotransduction, Cellular / Q000235:genetics; D009154:Mutation; D010812:Physical Stimulation; D014110:Touch
chemical_list: D029721:Drosophila Proteins