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In-situ Analysis of Cellular Poly(ADP-ribose) Production in Scrapie-infected Mouse Neuroblastoma Cells

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Abstract

Transmissible spongiform encephalopathies (TSEs), also called prion diseases, are characterized by formation of the disease-associated isoform of prion protein (PrPSc), which arises from a normal isoform termed PrPc by a post-translational conversion process occurring in an autocatalytic fashion. Oxidative stress has been proposed as a pathogenetic mechanism in TSEs and increased lipid peroxidation has recently been described in prion-infected cell cultures, suggesting an intrinsic link between the presence of prions and oxidative stress. We investigated if poly(ADP-ribose) formation can be detected in cultured cells upon prion infection, as this NAD+-consuming and DNA strand break-activated nuclear enzymatic reaction has the potential to cause rapid and lethal NAD+ depletion in cells under severe oxidative stress. Poly(ADP-ribose) production was analysed by immunofluorescence in freshly scrapie-infected Neuro2a–D11 mouse neuroblastoma cells, which had been confirmed by immunocytochemistry to produce PrPSc, and in uninfected controls. No spontaneous poly(ADP-ribose) specific signals were observed in infected or in uninfected cells, while both cell types readily reacted to H2O2 treatment with poly(ADP-ribose) synthesis in a dose-dependent manner, with no obvious difference in staining intensity at any dose tested. In summary, our data reveal that replication of scrapie agent in neuroblastoma cells can proceed without detectable stimulation of the cellular poly(ADP-ribosyl)ation system.

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Author notes

  1. Alexander Bürkle

    Present address: Lehrstuhl Molekulare Toxikologie, Fachbereich Biologie, Universität Konstanz, Fach X911, D-78457, Konstanz, Germany

Authors and Affiliations

  1. School of Clinical Medical Sciences – Gerontology, Institute for Ageing and Health, University of Newcastle upon Tyne, Wolfson Research Centre, NGH, Westgate Road, Newcastle-upon-Tyne, NE4 6BE, UK

    Yonghua Zhang & Alexander Bürkle

  2. Poly(ADP-ribose) Metabolism Laboratory, Health and Environment Unit, Laval University Medical Research Center, CHUQ, Faculty of Medicine, Laval University, 2705 Laurier Boulevard, Ste-Foy, Québec, Canada, G1V 4G2

    Guy G. Poirier

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  1. Yonghua Zhang
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Correspondence to Alexander Bürkle.

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Zhang, Y., Poirier, G.G. & Bürkle, A. In-situ Analysis of Cellular Poly(ADP-ribose) Production in Scrapie-infected Mouse Neuroblastoma Cells. Histochem J 34, 357–363 (2002). https://doi.org/10.1023/A:1023398130945

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