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Toxicological Sciences 59, 278-290 (2001)
Copyright © 2001 by the Society of Toxicology

NEUROTOXICOLOGY

Effects of Methylmercury and Mercuric Chloride on Differentiation and Cell Viability in PC12 Cells

D. K. Parran*, W. R. Mundy{dagger} and S. Barone, Jr.{dagger},1

* Curriculum in Toxicology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina; and {dagger} Cellular and Molecular Toxicology Branch, Neurotoxicology Division, Mail Drop 74-B, National Health and Environmental Effects Research Laboratory (NHEERL), U.S. Environmental Protection Agency, Research Triangle Park, North Carolina 27711

The effects of methylmercury (CH3Hg) or mercuric chloride (HgCl2) on neurite outgrowth and cell viability were quantified using undifferentiated (unprimed) and differentiated (primed) pheochromocytoma (PC12) cells. In unprimed cells, following 24-h exposure, CH3Hg significantly decreased NGF-stimulated neurite outgrowth at concentrations of 0.3–3 µM. However, HgCl2 significantly increased both neurite outgrowth and the number of branch points, a component of neurite outgrowth. In primed PC12 cells, following 24-h exposure, both CH3Hg and HgCl2 inhibited NGF-stimulated neurite outgrowth with an EC50 of approximately 0.03 µM; however, there was a difference between CH3Hg and HgCl2 effects on the subcomponents of total neurite outgrowth. CH3Hg significantly decreased both the number of branch points (0.3 µM) and fragment length (0.01 µM), while HgCl2 only decreased fragment length (0.03 µM). Cell viability was assessed in the same cultures by trypan-blue exclusion. In unprimed cells, the EC50 for cytotoxicity of CH3Hg in the presence and absence of NGF was 0.21 ± 0.04 and 0.87 ± 0.12 µM, respectively, and for HgCl2 in the presence and absence of NGF was 8.18 ± 1.52 and 5.02 ± 0.74 µM, respectively. In primed cells, the EC50 for cytotoxicity of CH3Hg in the presence or absence of NGF was 1.17 ± 0.38 and 0.73 ± 0.14 µM, respectively, and for HgCl2 in the presence or absence of NGF was 3.96 ± 0.82 and 3.81 ± 0.91 µM, respectively. In the primed PC12 model, cytotoxicity occurred at concentrations that were at least 30-fold higher than the EC50 for neurite outgrowth, suggesting that the mercurial compounds can act selectively on the process of differentiation.

Key Words: neurite outgrowth; developmental neurotoxicity; nerve growth factor (NGF).


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