Advisor Information
Bruce Chase
Location
Dr. C.C. and Mabel L. Criss Library
Presentation Type
Poster
Start Date
3-3-2017 2:15 PM
End Date
3-3-2017 3:30 PM
Abstract
Immunohistochemistry (IHC) is a useful research tool used to localize specific antigens in tissue sections with labeled antibodies based on antigen-antibody interactions. To obtain a clearer understanding of the cellular and subcellular localization of the Mind-Meld (MMD) protein during the developing Drosophila melanogaster embryo double indirect immunofluorescence was used to colocalize MMD with other proteins found in the fruit fly. An affinity-purified antibody raised in rabbit identifying all MMD isoforms was used with the fluorophore-labeled secondary antibody Alexa Fluor 594 specific to rabbit IgG. Well-characterized murine monoclonal antibodies with known subcellular localization and function in Drosophila: Fascicilin II (FASII), Short stop (SHOT), β-Tubulin, Spaghetti squash (SQH), and Neurotactin (NRT) were used with the fluorophore-labeled secondary antibody Alexa Fluor 488 specific to mouse IgG. All fixed embryos were stained with the nuclear marker DAPI to identify the density and arrangement of the nuclei and the F-actin marker Phalloidin. Confocal microscopy which provides three-dimensional optical resolution was used to visualize the localization of the proteins. In the present work, it is demonstrated that MMD colocalizes with SHOT, β-Tubulin, Phalloidin and SQH suggesting MMD’s importance in cell adhesion, cell migration and establishing the cytoskeletal. The characterization of the cellular and subcellular localization of the MMD protein during the developing Drosophila will provide insight into the context in which mmd functions in human disease processes.
Functional Genetics of Mind-Meld in Drosophila Melanogaster
Dr. C.C. and Mabel L. Criss Library
Immunohistochemistry (IHC) is a useful research tool used to localize specific antigens in tissue sections with labeled antibodies based on antigen-antibody interactions. To obtain a clearer understanding of the cellular and subcellular localization of the Mind-Meld (MMD) protein during the developing Drosophila melanogaster embryo double indirect immunofluorescence was used to colocalize MMD with other proteins found in the fruit fly. An affinity-purified antibody raised in rabbit identifying all MMD isoforms was used with the fluorophore-labeled secondary antibody Alexa Fluor 594 specific to rabbit IgG. Well-characterized murine monoclonal antibodies with known subcellular localization and function in Drosophila: Fascicilin II (FASII), Short stop (SHOT), β-Tubulin, Spaghetti squash (SQH), and Neurotactin (NRT) were used with the fluorophore-labeled secondary antibody Alexa Fluor 488 specific to mouse IgG. All fixed embryos were stained with the nuclear marker DAPI to identify the density and arrangement of the nuclei and the F-actin marker Phalloidin. Confocal microscopy which provides three-dimensional optical resolution was used to visualize the localization of the proteins. In the present work, it is demonstrated that MMD colocalizes with SHOT, β-Tubulin, Phalloidin and SQH suggesting MMD’s importance in cell adhesion, cell migration and establishing the cytoskeletal. The characterization of the cellular and subcellular localization of the MMD protein during the developing Drosophila will provide insight into the context in which mmd functions in human disease processes.
Additional Information (Optional)
Winner of Outstanding Graduate Poster Presentation