title: The role of the glutamatergic NMDA receptor in nanosilver-evoked neurotoxicity in primary cultures of cerebellar granule cells.
abstract: Nanoparticles are known to enter the vertebrate brain, but little is known about their neurotoxicity. The aim of this study is to investigate mechanisms of the contribution of AgNPs to neuronal cell death using primary cultures of rat cerebellar granule cells (CGCs). We tested the role of glutamatergic N-methyl-d-aspartate receptors (NMDA) in AgNP-evoked neurotoxicity using MK-801, a noncompetitive inhibitor of NMDAR. We used commercially available 0.2% PVP-coated AgNPs <100 nm in a concentration range of 2.5-75 μg/ml sonicated with fetal calf serum. After a 10 min incubation period, a dose-dependent increase in the uptake of (45)Ca(2+) into neurons was observed in the presence of 25-75 μg/mL AgNPs which was completely abolished by addition of MK-801. Using the fluorescent dye fluo3 AM we observed an increase in the intracellular calcium level by 87% compared to control. ROS production was found to increase by about 30% over control after a 30-min incubation with 75 μg/mL AgNPs. Further, we observed a significant decrease in the mitochondrial potential during a 30-min incubation with AgNPs. Administration of MK-801 was found to provide a protective effect. Our results show that excitotoxicity via activation of NMDA receptor, followed by calcium imbalance, destabilization of mitochondrial function and ROS production, indicate an important mechanism involved in neurotoxicity evoked by AgNPs in cultured neurons.
authors.affiliation: Laboratory of Pharmaconeurochemistry, Department of Neurochemistry, Mossakowski Medical Research Centre, Polish Academy of Sciences, Pawińskiego 5, 02-106 Warsaw, Poland.
authors.forename: Elżbieta
authors.initials: E
authors.lastname: Ziemińska
authors.affiliation: Laboratory of Pharmaconeurochemistry, Department of Neurochemistry, Mossakowski Medical Research Centre, Polish Academy of Sciences, Pawińskiego 5, 02-106 Warsaw, Poland.
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Stafiej
authors.affiliation: Laboratory of Pathoneurochemistry, Department of Neurochemistry, Mossakowski Medical Research Centre, Polish Academy of Sciences, Pawińskiego 5, 02-106 Warsaw, Poland. Electronic address: lidkas@imdik.pan.pl.
authors.forename: Lidia
authors.initials: L
authors.lastname: Strużyńska
mesh_terms: D000814:Aniline Compounds / Q000737:chemistry; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000187:drug effects* / Q000473:pathology; D016291:Dizocilpine Maleate / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005260:Female; D005456:Fluorescent Dyes / Q000737:chemistry; D008297:Male; D053768:Metal Nanoparticles / Q000008:administration & dosage / Q000633:toxicity*; D008928:Mitochondria / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000473:pathology; D020258:Neurotoxicity Syndromes / Q000209:etiology* / Q000503:physiopathology; D051381:Rats; D017208:Rats, Wistar; D017382:Reactive Oxygen Species / Q000378:metabolism; D016194:Receptors, N-Methyl-D-Aspartate / Q000187:drug effects / Q000378:metabolism*; D044967:Serum; D012834:Silver / Q000008:administration & dosage / Q000633:toxicity*; D014966:Xanthenes / Q000737:chemistry
chemical_list: D000814:Aniline Compounds; D005456:Fluorescent Dyes; D017382:Reactive Oxygen Species; D016194:Receptors, N-Methyl-D-Aspartate; D014966:Xanthenes; C059715:Fluo-3; D012834:Silver; D016291:Dizocilpine Maleate; D002118:Calcium
keywords: Calcium homeostasis; Excitotoxicity; MK-801; Nanosilver; Nanotoxicity; Reactive oxygen species