Dissection and Immunofluorescent Staining of Mushroom Body and Photoreceptor Neurons in Adult Drosophila melanogaster Brains
| Autor: | Seth M. Kelly, Michael Kahl, Alexandra R Elchert |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Nervous system General Immunology and Microbiology biology General Chemical Engineering General Neuroscience fungi MARCM Neurogenesis Synaptogenesis biology.organism_classification General Biochemistry Genetics and Molecular Biology 03 medical and health sciences 030104 developmental biology medicine.anatomical_structure nervous system Mushroom bodies medicine Axon guidance Drosophila melanogaster Drosophila Neuroscience |
| Zdroj: | Journal of Visualized Experiments. |
| ISSN: | 1940-087X |
| Popis: | Nervous system development involves a sequential series of events that are coordinated by several signaling pathways and regulatory networks. Many of the proteins involved in these pathways are evolutionarily conserved between mammals and other eukaryotes, such as the fruit fly Drosophila melanogaster, suggesting that similar organizing principles exist during the development of these organisms. Importantly, Drosophila has been used extensively to identify cellular and molecular mechanisms regulating processes that are required in mammals including neurogenesis, differentiation, axonal guidance, and synaptogenesis. Flies have also been used successfully to model a variety of human neurodevelopmental diseases. Here we describe a protocol for the step-by-step microdissection, fixation, and immunofluorescent localization of proteins within the adult Drosophila brain. This protocol focuses on two example neuronal populations, mushroom body neurons and retinal photoreceptors, and includes optional steps to trace individual mushroom body neurons using Mosaic Analysis with a Repressible Cell Marker (MARCM) technique. Example data from both wild-type and mutant brains are shown along with a brief description of a scoring criteria for axonal guidance defects. While this protocol highlights two well-established antibodies for investigating the morphology of mushroom body and photoreceptor neurons, other Drosophila brain regions and the localization of proteins within other brain regions can also be investigated using this protocol. |
| Databáze: | OpenAIRE |
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