An optimized method for histological detection of dopaminergic neurons in Drosophila melanogaster.
Parkinson's disease (PD) affects >1 million Americans and is marked by the loss of dopaminergic neurons in the substantia nigra. PD has been linked to two causative factors: genetic risks (hereditary PD) and environmental toxins (idiopathic PD). In recent years, considerable effort has been devoted to the development of a Drosophila model of human PD that might be useful for examining the cellular mechanisms of PD pathology by genetic screening. In 2000, Feany and Bender reported a Drosophila model of PD in which transgenic flies expressing human mutant alpha-synuclein exhibited shortened life spans, dopaminergic losses, Parkinsonian behaviors, and Lewy bodies in surviving dopaminergic neurons. Since then, a number of studies have been published that validate the model or build on it; conversely, a number report an inability to replicate the results and suggest that most protocols for dopaminergic histology underreport the actual numbers of dopaminergic neurons in the insect brain. Here we report the optimization of dopaminergic histology in Drosophila and identification of new dopaminergic neurons, show the remarkable dendritic complexity of these neurons, and provide an updated count of these neurons in adult brains. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials.
Eccles Institute of Human Genetics, University of Utah, 15 North 2030 East, Salt Lake City, UT 84112, USA.
Daria
D
Drobysheva
Kristen
K
Ameel
Brandon
B
Welch
Esther
E
Ellison
Khan
K
Chaichana
Bryan
B
Hoang
Shilpy
S
Sharma
Wendi
W
Neckameyer
Irina
I
Srinakevitch
Kelley J
KJ
Murphy
Aloisia
A
Schmid
D000818:Animals; D001921:Brain / Q000166:cytology / Q000378:metabolism; D002452:Cell Count; D003712:Dendrites / Q000648:ultrastructure; D004298:Dopamine / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000166:cytology / Q000378:metabolism*; D005404:Fixatives; D016591:Histocytological Preparation Techniques / Q000379:methods*; D007150:Immunohistochemistry; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D013194:Staining and Labeling; D016610:Tissue Embedding; D016707:Tissue Fixation
D005404:Fixatives; D004298:Dopamine