title: JNK3-mediated apoptotic cell death in primary dopaminergic neurons.
abstract: Investigation of mechanisms responsible for dopaminergic neuron death is critical for understanding the pathogenesis of Parkinson's disease, yet this is often quite challenging technically. Here, we describe detailed methods for culturing primary mesencephalic dopaminergic neurons and examining the activation of c-Jun N-terminal protein Kinase (JNK) in these cultures. We utilized immunocytochemistry and computerized analysis to quantify the number of surviving dopaminergic neurons and JNK activation in dopaminergic neurons. TUNEL staining was used to quantify apoptotic cell death. siRNA was used to specifically inhibit JNK3, the neural specific isoform of JNK. Our data implicate the activation of JNK3 in rotenone-induced dopaminergic neuron apoptosis.
authors.affiliation: Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, WA, USA.
authors.forename: Won-Seok
authors.initials: WS
authors.lastname: Choi
authors.forename: Heather M
authors.initials: HM
authors.lastname: Klintworth
authors.forename: Zhengui
authors.initials: Z
authors.lastname: Xia
mesh_terms: D000818:Animals; D017209:Apoptosis; D053148:Caspase 3 / Q000378:metabolism; D059290:Dopaminergic Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D004622:Embryo, Mammalian / Q000166:cytology; D004789:Enzyme Activation; D005260:Female; D020287:In Situ Nick-End Labeling; D008636:Mesencephalon / Q000166:cytology; D051379:Mice; D048057:Mitogen-Activated Protein Kinase 10 / Q000235:genetics / Q000378:metabolism*; D010269:Paraquat / Q000494:pharmacology; D010766:Phosphorylation; D011247:Pregnancy; D061251:Primary Cell Culture; D034622:RNA Interference; D051381:Rats; D017207:Rats, Sprague-Dawley; D012402:Rotenone / Q000494:pharmacology; D014162:Transfection
chemical_list: D012402:Rotenone; D048057:Mitogen-Activated Protein Kinase 10; C505410:Casp3 protein, rat; D053148:Caspase 3; D010269:Paraquat