title: Assessing neurodegenerative phenotypes in Drosophila dopaminergic neurons by climbing assays and whole brain immunostaining.
abstract: Drosophila melanogaster is a valuable model organism to study aging and pathological degenerative processes in the nervous system. The advantages of the fly as an experimental system include its genetic tractability, short life span and the possibility to observe and quantitatively analyze complex behaviors. The expression of disease-linked genes in specific neuronal populations of the Drosophila brain, can be used to model human neurodegenerative diseases such as Parkinson's and Alzheimer's (5). Dopaminergic (DA) neurons are among the most vulnerable neuronal populations in the aging human brain. In Parkinson's disease (PD), the most common neurodegenerative movement disorder, the accelerated loss of DA neurons leads to a progressive and irreversible decline in locomotor function. In addition to age and exposure to environmental toxins, loss of DA neurons is exacerbated by specific mutations in the coding or promoter regions of several genes. The identification of such PD-associated alleles provides the experimental basis for the use of Drosophila as a model to study neurodegeneration of DA neurons in vivo. For example, the expression of the PD-linked human α-synuclein gene in Drosophila DA neurons recapitulates some features of the human disease, e.g. progressive loss of DA neurons and declining locomotor function (2). Accordingly, this model has been successfully used to identify potential therapeutic targets in PD (8). Here we describe two assays that have commonly been used to study age-dependent neurodegeneration of DA neurons in Drosophila: a climbing assay based on the startle-induced negative geotaxis response and tyrosine hydroxylase immunostaining of whole adult brain mounts to monitor the number of DA neurons at different ages. In both cases, in vivo expression of UAS transgenes specifically in DA neurons can be achieved by using a tyrosine hydroxylase (TH) promoter-Gal4 driver line (3, 10).
authors: Department of Biomolecular Genetics, University of Rochester Medical Center.
authors: Maria Cecilia
authors: MC
authors: Barone
authors: Dirk
authors: D
authors: Bohmann
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism / Q000473:pathology*; D059290:Dopaminergic Neurons / Q000378:metabolism / Q000473:pathology*; D004331:Drosophila melanogaster; D005260:Female; D005455:Fluorescent Antibody Technique / Q000379:methods; D006801:Humans; D008297:Male; D019636:Neurodegenerative Diseases / Q000378:metabolism / Q000473:pathology*; D010641:Phenotype; D014446:Tyrosine 3-Monooxygenase / Q000737:chemistry; D051844:alpha-Synuclein / Q000235:genetics
chemical_list: D051844:alpha-Synuclein; D014446:Tyrosine 3-Monooxygenase
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