Role of glutathione in determining the differential sensitivity between the cortical and cerebellar regions towards mercury-induced oxidative stress.
Certain discrete areas of the CNS exhibit enhanced sensitivity towards MeHg. To determine whether GSH is responsible for this particular sensitivity, we investigated its role in MeHg-induced oxidative insult in primary neuronal and astroglial cell cultures of both cerebellar and cortical origins. For this purpose, ROS and GSH were measured with the fluorescent indicators, CMH(2)DCFDA and MCB. Cell associated-MeHg was measured with (14)C-radiolabeled MeHg. The intracellular GSH content was modified by pretreatment with NAC or DEM. For each of the dependent variables (ROS, GSH, and MTT), there was an overall significant effect of cellular origin, MeHg and pretreatment in all the cell cultures. A trend towards significant interaction between originxMeHgxpretreatment was observed only for the dependent variable, ROS (astrocytes p=0.056; neurons p=0.000). For GSH, a significant interaction between originxMeHg was observed only in astrocytes (p=0.030). The cerebellar cell cultures were more vulnerable (astrocytes(mean)=223.77; neurons(mean)=138.06) to ROS than the cortical cell cultures (astrocytes(mean)=125.18; neurons(mean)=107.91) for each of the tested treatments. The cell associated-MeHg increased when treated with DEM, and the cerebellar cultures varied significantly from the cortical cultures. Non-significant interactions between originxMeHgxpretreatment for GSH did not explain the significant interactions responsible for the increased amount of ROS produced in these cultures. In summary, although GSH modulation influences MeHg-induced toxicity, the difference in the content of GSH in cortical and cerebellar cultures fails to account for the increased ROS production in cerebellar cultures. Hence, different approaches for the future studies regarding the mechanisms behind selectivity of MeHg have been discussed.
Kaur
Parvinder
P
Department of Neuroscience, Norwegian University of Science and Technology, N-7489 Trondheim, Norway.
Aschner
Michael
M
Syversen
Tore
T
D000818:Animals; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D016923:Cell Death / Q000187:drug effects; D002531:Cerebellum / Q000187:drug effects* / Q000378:metabolism* / Q000473:pathology; D002540:Cerebral Cortex / Q000187:drug effects* / Q000378:metabolism* / Q000473:pathology; D005456:Fluorescent Dyes / Q000737:chemistry; D005562:Formazans / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism*; D020262:Mercury Poisoning, Nervous System / Q000378:metabolism / Q000473:pathology; D008767:Methylmercury Compounds / Q000633:toxicity*; D051379:Mice; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D018384:Oxidative Stress / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism; D013778:Tetrazolium Salts / Q000378:metabolism
D005456:Fluorescent Dyes; D005562:Formazans; D008767:Methylmercury Compounds; D017382:Reactive Oxygen Species; D013778:Tetrazolium Salts; C079782:MTT formazan; D005978:Glutathione; C004925:methylmercuric chloride