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.
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Acknowledgments
This work was supported by the Post-doctoral Fellowship Program of KOSEF (WSC), Environmental Pathology/Toxicology Training Grant T32 ES007032 (HMK) and NIH Grants ES012215 and ES013696 (ZX).
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Choi, WS., Klintworth, H.M., Xia, Z. (2011). JNK3-Mediated Apoptotic Cell Death in Primary Dopaminergic Neurons. In: Costa, L., Giordano, G., Guizzetti, M. (eds) In Vitro Neurotoxicology. Methods in Molecular Biology, vol 758. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-170-3_19
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DOI: https://doi.org/10.1007/978-1-61779-170-3_19
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