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Optimized dopaminergic neuron immunostaining in Drosophila melanogaster
Author(s) -
Murphy Kelley J,
Drobysheva Daria,
Ameel Kristen,
Ellison Esther,
Hoang Bryan,
Schmid Aloisia
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.2_supplement.257
Subject(s) - fixative , drosophila melanogaster , dopaminergic , immunostaining , biology , vibratome , pathology , immunolabeling , disease , immunohistochemistry , parkinson's disease , neuroscience , epitope , medicine , microbiology and biotechnology , immunology , dopamine , antibody , gene , genetics , cytoplasm
Parkinson's Disease (PD) affects more than 1 million Americans and 50,000 new cases are diagnosed in the US each year. The vast majority of PD cases have been shown to be idiopathic and caused by environmental toxins; therefore creating an animal model to test both whether a given toxin is causative of the disease or not, and what drugs can be administered to ameliorate the effects of those toxins would advance prevention and treatment of PD in humans. The remaining 10% of the human cases of PD are caused by genetic factors. Drosophila melanogaster provides us with an excellent model organism for both idiopathic and genetic disease modeling, but we are faced with the challenge of comparing different studies and their different reports of dopaminergic neuronal loss (the diagnostic hallmark of PD in humans). We have developed an optimized protocol of fixation, embedding, and immunohistochemistry in the fly brain to demonstrate those neurons. This protocol combines a weak cross‐linking fixative containing an anti‐oxidant to provide protection to the tissue, microwave‐enhanced penetration of the fixative into the fly brain, thick agarose sectioning with a vibratome, and immunostaining using an antibody specific to the Drosophila ortholog of a dopaminergic epitope to identify these neurons.