title: Changed iron regulation in scrapie-infected neuroblastoma cells.
abstract: Prion diseases are characterized by the conversion of the normal cellular prion protein PrP(C) into a pathogenic isoform, PrP(Sc). The mechanisms involved in neuronal cell death in prion diseases are largely unknown, but accumulating evidence has demonstrated oxidative impairment along with metal imbalances in scrapie-infected brains. In this study, we report changes in cellular iron metabolism in scrapie-infected mouse neuroblastoma N2a cells (ScN2a). We detected twofold lower total cellular iron and calcein-chelatable cytosolic labile iron pool (LIP) in ScN2a cells as compared to the N2a cells. We also measured in ScN2a cells significantly lower activities of iron regulatory proteins 1 and 2 (IRP1 and IRP2, respectively), regulators of cellular iron by sensing cytosolic free iron levels and controlling posttranscriptionally the expression of the major iron transport protein transferrin receptor 1 (TfR1) and the iron sequestration protein ferritin. IRP1 and IRP2 protein levels were decreased by 40% and 50%, respectively, in ScN2a cells. TfR1 protein levels were fourfold reduced and ferritin levels were threefold reduced in ScN2a cells. TfR1 and ferritin mRNA levels were significantly reduced in ScN2a cells. ScN2a cells responded normally to iron and iron chelator treatment with respect to the activities of IRP1 and IRP2, and biosynthesis of TfR1 and ferritin. However, the activities of IRP1 and IRP2, and protein levels of TfR1 and ferritin, were still significantly lower in iron-depleted ScN2a cells as compared to the N2a cells, suggesting lower need for iron in ScN2a cells. Our results demonstrate that scrapie infection leads to changes in cellular iron metabolism, affecting both total cellular and cytosolic free iron, and the activities and expression of major regulators of cellular iron homeostasis.
authors.affiliation: Department of Neurochemistry and Neurotoxicology, Stockholm University, S-106 91 Stockholm, Sweden.
authors.forename: Sandra
authors.initials: S
authors.lastname: Fernaeus
authors.forename: Jonas
authors.initials: J
authors.lastname: Hälldin
authors.forename: Katarina
authors.initials: K
authors.lastname: Bedecs
authors.forename: Tiit
authors.initials: T
authors.lastname: Land
mesh_terms: D000818:Animals; D015139:Blotting, Southern / Q000379:methods; D015153:Blotting, Western / Q000379:methods; D045744:Cell Line, Tumor; D003676:Deferoxamine / Q000494:pharmacology; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods; D005290:Ferric Compounds / Q000494:pharmacology; D005293:Ferritins / Q000235:genetics / Q000378:metabolism; D005452:Fluoresceins / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D007239:Infections; D007501:Iron / Q000378:metabolism*; D035941:Iron Regulatory Protein 1 / Q000378:metabolism; D035942:Iron Regulatory Protein 2 / Q000378:metabolism; D051379:Mice; D009447:Neuroblastoma / Q000378:metabolism*; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D011990:Receptors, Transferrin / Q000235:genetics / Q000378:metabolism; D012608:Scrapie / Q000378:metabolism*; D013997:Time Factors
chemical_list: D005290:Ferric Compounds; D005452:Fluoresceins; D000644:Quaternary Ammonium Compounds; D012333:RNA, Messenger; D011990:Receptors, Transferrin; C085925:calcein AM; D005293:Ferritins; D007501:Iron; D035941:Iron Regulatory Protein 1; D035942:Iron Regulatory Protein 2; D003676:Deferoxamine; C013531:ferric ammonium citrate