Journal
DEVELOPMENTAL NEUROSCIENCE
Volume 24, Issue 2-3, Pages 125-133Publisher
KARGER
DOI: 10.1159/000065691
Keywords
zinc deficiency; iron toxicity; IMR-32 neuroblastoma cells; oxidative stress; apoptosis
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Funding
- EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH &HUMAN DEVELOPMENT [R01HD001743] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES [P30DK035747] Funding Source: NIH RePORTER
- NICHD NIH HHS [HD01743] Funding Source: Medline
- NIDDK NIH HHS [DK35747] Funding Source: Medline
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The current work tested the hypothesis that the zinc status of a cell influences its sensitivity to iron-induced oxidative stress. Human IMR-32 neuroblastoma cells were cultured for 24 h in nonchelated control media (5 muM zinc; 4.5 muM iron), or in media that was treated with DTPA to reduce its zinc content (chelated media). Chelated media was supplemented with zinc to achieve concentrations of 1.5-50 muM Zn. The media was then replaced with serum-free complex media (0.9 muM Zn) with either no added iron (0.6 muM Fe), or iron (FeCl3) added at concentrations ranging from 15 to 100 muM. Cells were cultured for an additional 3- to 24-hour period. Over the 24-hour period, cells cultured in the control iron media had good viability, and they displayed the gross morphology typical of these cells in culture. With 100 muM iron, cell viability was low in all groups. After 24 h and at iron concentrations between 15-50 muM, cells that had been cultured in the low zinc-chelated media (1.5 muM Zn) showed a concentration-dependent increase in 5 (or 6)-carboxy2'7'-dichlorodihydrofluorescein diacetate (DCDCDHF) fluorescence (oxidative stress) and decrease in cell viability. A positive correlation between both parameters was observed (r = 0.92). These cells had altered morphology and high level of nucleosomes suggestive of cell death by apoptosis. These results support the concept that the zinc status of IMR-32 neuroblastoma cells modulates their sensitivity to iron overload. Copyright (C) 2002 S. Karger AG, Basel.
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