Removal of Drosophila Muscle Tissue from Larval Fillets for Immunofluorescence Analysis of Sensory Neurons and Epidermal Cells.

TitleRemoval of Drosophila Muscle Tissue from Larval Fillets for Immunofluorescence Analysis of Sensory Neurons and Epidermal Cells.
Publication TypeJournal Article
Year of Publication2016
AuthorsTenenbaum, CM, Gavis, ER
JournalJ Vis Exp
Issue117
Date Published2016 Nov 02
ISSN1940-087X
KeywordsAnimals, Dendrites, Drosophila, Drosophila Proteins, Fluorescent Antibody Technique, Larva, Models, Animal, Morphogenesis, Muscles, Neurons, Sensory Receptor Cells
Abstract

<p>Drosophila larval dendritic arborization (da) neurons are a popular model for investigating mechanisms of neuronal morphogenesis. Da neurons develop in communication with the epidermal cells they innervate and thus their analysis benefits from in situ visualization of both neuronally and epidermally expressed proteins by immunofluorescence. Traditional methods of preparing larval fillets for immunofluorescence experiments leave intact the muscle tissue that covers most of the body wall, presenting several challenges to imaging neuronal and epidermal proteins. Here we describe a method for removing muscle tissue from Drosophila larval fillets. This protocol enables imaging of proteins that are otherwise obscured by muscle tissue, improves signal to noise ratio, and facilitates the use of super-resolution microscopy to study da neuron development.</p>

DOI10.3791/54670
Alternate JournalJ Vis Exp
PubMed ID27842373
PubMed Central IDPMC5226124
Grant ListR01 GM061107 / GM / NIGMS NIH HHS / United States
R01 GM067758 / GM / NIGMS NIH HHS / United States