NDLI: Toxicity of 6-hydroxydopamine: live cell imaging of cytoplasmic redox flux

Content Provider	Springer Nature Link
Author	Dooley, Colette T. Li, Ling Misler, Jaime A. Thompson, Jane H.
Copyright Year	2012
Abstract	Oxidative stress contributes to several debilitating neurodegenerative diseases. To facilitate direct monitoring of the cytoplasmic oxidation state in neuronal cells, we have developed roTurbo by including several mutations: F223R, A206K, and six of the mutations for superfolder green fluorescent protein. Thus we have generated an improved redox sensor that is much brighter in cells and oxidizes more readily than roGFP2. Cytoplasmic expression of the sensor demonstrated the temporal pattern of 6-hydroxydopamine (6-OHDA) induced oxidative stress in a neuroblastoma cell line (SH-SY5Y). Two distinct oxidation responses were identified in SH-SY5Y cells but a single response observed in cells lacking monoamine transporters (HEK293). While both cell lines exhibited a rapid transient oxidation in response to 6-OHDA, a second oxidative response coincident with cell death was observed only in SH-SY5Y cells, indicating an intracellular metabolism of 6-OHDA, and or its metabolites are involved. In contrast, exogenously applied hydrogen peroxide induced a cellular oxidative response similar to the first oxidation peak, and cell loss was minimal. Glucose deprivation enhanced the oxidative stress induced by 6-OHDA, confirming the pivotal role played by glucose in maintaining a reduced cytoplasmic environment. While these studies support previous findings that catecholamine auto-oxidation products cause oxidative stress, our findings also support studies indicating 6-OHDA induces lethal oxidative stress responses unrelated to production of hydrogen peroxide. Finally, temporal imaging revealed the sporadic nature of the toxicity induced by 6-OHDA in neuroblastoma cells.
Starting Page	89
Ending Page	101
Page Count	13
File Format	PDF
ISSN	07422091
Journal	Cell Biology and Toxicology
Volume Number	28
Issue Number	2
e-ISSN	15736822
Language	English
Publisher	Springer Netherlands
Publisher Date	2012-01-08
Publisher Place	Dordrecht
Access Restriction	Subscribed
Subject Keyword	Dopamine Green fluorescent protein Hydrogen peroxide Oxidative stress Redox Neuroblastoma Pharmacology/Toxicology Cell Biology Biochemistry
Content Type	Text
Resource Type	Article
Subject	Cell Biology Health, Toxicology and Mutagenesis Toxicology

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