Journal
CHEMOSPHERE
Volume 74, Issue 1, Pages 112-118Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.chemosphere.2008.09.019
Keywords
Demethylation; Brain; Methylmercury; Inorganic mercury toxicity; Astrocytes
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Funding
- National Sciences and Engineering Research Council (NSERC) of Canada Discovery Research
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Mercury (Hg) is a well-known neurotoxicant but its toxicity depends on the species present. A steady emergence of inorganic Hg in the brain following chronic and accidental exposure to methylmercury (MeHg) has suggested that MeHg can undergo demethylation. The objective of this study is to develop an in vitro model to study factors affecting Hg demethylation in the central nervous system. Astrocytes obtained from neonatal rat pups were cultured for 24 h with 1 mu M MeHg in the presence of two pro-oxidants, buthionine sulphoximine (BSO) and rotenone. The BSO treatment produced a 21% increase in reactive oxygen species (ROS) content compared to the control (control vs. BSO; 100 +/- 1.35 vs. 121 +/- 1.52 relative fluorescence units (RFU)mg(-1) protein. p < 0.001) but did not affect total Hg accumulation (control vs. BSO = 86.5 +/- 4.14 ng mg(-1) vs. 95.7 +/- 9.26 ng mg(-1)). Rotenone increased ROS levels 107% (control vs. rotenone; 100%+/- 1.35 vs. 207%+/- 6.78 RFU mg(-1) protein, p < 0.001) and significantly increased the accumulation of total Hg (control vs. rotenone = 86.5 +/- 4.14 ng mg(-1) vs. 124 +/- 3.80 ng mg(-1), p < 0.001). There was no detectable demethylation in the control or BSO treated group, however, the rotenone treatment significantly increased the demethylation (control vs. rotenone = -1.86 +/- 5.57% vs. 16.3 +/- 2.68%, p < 0.05). For the first time, we have demonstrated in an in vitro primary astrocyte culture model that MeHg can be converted to inorganic Hg and demethylation increases with oxidative stress. Our results provide a useful model to study demethylation of Hg in astrocytes and to explore potential ways to protect against Hg toxicity. (c) 2008 Elsevier Ltd. All rights reserved.
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