title: Infectious prion protein alters manganese transport and neurotoxicity in a cell culture model of prion disease.
abstract: Protein misfolding and aggregation are considered key features of many neurodegenerative diseases, but biochemical mechanisms underlying protein misfolding and the propagation of protein aggregates are not well understood. Prion disease is a classical neurodegenerative disorder resulting from the misfolding of endogenously expressed normal cellular prion protein (PrP(C)). Although the exact function of PrP(C) has not been fully elucidated, studies have suggested that it can function as a metal binding protein. Interestingly, increased brain manganese (Mn) levels have been reported in various prion diseases indicating divalent metals also may play a role in the disease process. Recently, we reported that PrP(C) protects against Mn-induced cytotoxicity in a neural cell culture model. To further understand the role of Mn in prion diseases, we examined Mn neurotoxicity in an infectious cell culture model of prion disease. Our results show CAD5 scrapie-infected cells were more resistant to Mn neurotoxicity as compared to uninfected cells (EC(50)=428.8 μM for CAD5 infected cells vs. 211.6 μM for uninfected cells). Additionally, treatment with 300 μM Mn in persistently infected CAD5 cells showed a reduction in mitochondrial impairment, caspase-3 activation, and DNA fragmentation when compared to uninfected cells. Scrapie-infected cells also showed significantly reduced Mn uptake as measured by inductively coupled plasma-mass spectrometry (ICP-MS), and altered expression of metal transporting proteins DMT1 and transferrin. Together, our data indicate that conversion of PrP to the pathogenic isoform enhances its ability to regulate Mn homeostasis, and suggest that understanding the interaction of metals with disease-specific proteins may provide further insight to protein aggregation in neurodegenerative diseases.
authors.affiliation: Department of Biomedical Sciences, Iowa Center for Advanced Neurotoxicity, Ames, IA 50011, USA.
authors.forename: Dustin P
authors.initials: DP
authors.lastname: Martin
authors.forename: Vellareddy
authors.initials: V
authors.lastname: Anantharam
authors.forename: Huajun
authors.initials: H
authors.lastname: Jin
authors.forename: Travis
authors.initials: T
authors.lastname: Witte
authors.forename: Robert
authors.initials: R
authors.lastname: Houk
authors.forename: Arthi
authors.initials: A
authors.lastname: Kanthasamy
authors.forename: Anumantha G
authors.initials: AG
authors.lastname: Kanthasamy
mesh_terms: D000818:Animals; D001692:Biological Transport / Q000502:physiology; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D008345:Manganese / Q000378:metabolism* / Q000633:toxicity*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D017096:Prion Diseases / Q000378:metabolism*; D011328:Prions / Q000378:metabolism*
chemical_list: D011328:Prions; D008345:Manganese