title: Acute exposure to prion infection induces transient oxidative stress progressing to be cumulatively deleterious with chronic propagation in vitro.
abstract: Neuronal loss is a pathological feature of prion diseases for which increased reactive oxygen species (ROS) and consequent oxidative stress is one proposed mechanism. The processes underlying ROS production in prion disease and the precise relationship to misfolding of the prion protein remain obscure. Using cell culture models of prion infection we found that cells demonstrate a rapid, prion protein (PrP) dependent, increase in intracellular ROS following exposure to infectious inoculum. ROS production correlated with internalisation and increased intracellular protease resistant PrP (PrP(Res)). The ROS increase was predominantly lysosomal in origin but not sustained, with cells adapting within 48 hours. Overall ROS levels remained normal in the chronically prion infected cell population; however a subpopulation characterised by loss of membrane phosphatidylserine asymmetry exhibited highly peroxidised intracellular aggregates that localised with PrP and intense caspase activation. These apoptotic cells showed increased ROS closely correlating with increased PrP(Res). Our findings demonstrate that a PrP-dependent, transient, increase in intracellular ROS is characteristic of acute cellular prion infection, while chronic phases of prion infection in vitro are associated with a significant subpopulation manifesting apoptosis accompanying heightened oxidative stress and increased PrP(Res) burden. Such observations strengthen the direct links between heightened ROS and ongoing prion propagation with eventual cellular demise.
authors.affiliation: Department of Pathology, The University of Melbourne, Parkville, 3010, Australia.
authors.forename: Cathryn L
authors.initials: CL
authors.lastname: Haigh
authors.forename: Amelia R
authors.initials: AR
authors.lastname: McGlade
authors.forename: Victoria
authors.initials: V
authors.lastname: Lewis
authors.forename: Colin L
authors.initials: CL
authors.lastname: Masters
authors.forename: Victoria A
authors.initials: VA
authors.lastname: Lawson
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Collins
mesh_terms: D000818:Animals; D017209:Apoptosis; D020169:Caspases / Q000378:metabolism*; D002460:Cell Line; D018450:Disease Progression; D006801:Humans; D051379:Mice; D023421:Models, Animal; D018384:Oxidative Stress; D018620:PrPSc Proteins / Q000378:metabolism*; D017096:Prion Diseases / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D011328:Prions / Q000378:metabolism* / Q000472:pathogenicity; D011817:Rabbits; D017382:Reactive Oxygen Species / Q000378:metabolism*
chemical_list: D018620:PrPSc Proteins; D011328:Prions; D017382:Reactive Oxygen Species; D020169:Caspases