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ARS Home Midwest Area Ames, Iowa National Animal Disease Center Virus and Prion Research Research Publications at this Location





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Title:   Author  item
                                            KONDRU, NAVEEN - Iowa State Universityitem
                                            Greenlee, Justinitem
                                            WEST GREENLEE, MARY - Iowa State Universityitem
                                            MANNE, SIREESHA - Iowa State Universityitem
                                            MAIN, ROGER - Iowa State Universityitem
                                            HALBUR, PATRICK - Iowa State Universityitem
                                            KANTHASAMY, ARTHI - Iowa State Universityitem
                                            KANTHASAMY, ANUMANTHA - Iowa State University
  Submitted to: Prion
                                    Publication Type: Abstract Only
                                    Publication Acceptance Date: 3/31/2015
                                    Publication  Date: 5/25/2015
                                    Citation: Kondru, N., Greenlee, J., Greenlee, H., Manne, S., Kong, Q., Halbur, P., Kanthasamy, A., Kanthasamy, A. 2015. Development and characterization of an ex-vivo brain slice culture model of chronic wasting disease. Prion 2015. p. S68.Interpretive Summary:
                                Technical Abstract: Prion diseases have long incubation times in vivo, therefore, modeling the diseases ex-vivo will advance the development of rationale-based therapeutic strategies. An organotypic slice culture assay (POSCA) was recently developed for scrapie prions by inoculating mouse cerebellar brain slices with RML scrapie. However, efforts to develop a POSCA model for CWD have been futile due to the species barrier between mouse and cervids. To overcome this difficulty, we adopted a transgenic cervidized (Tg12) mouse model and successfully developed an ex-vivo brain slice culture model for CWD. We prepared 350-μm cerebellar slices from Tg12 mouse pups and then incubated them with CWD brain homogenate and washed thoroughly. With the genotypes of the pups blinded, the cultures were grown over 42-48 days before being tested for CWD infectivity using the recently developed Real-time quaking-induced conversion (RT-QuIC) assay. Hamster PrP was used as a substrate for the assay. Slices from Tg12 cervidized pups with PrP+/- genotype tested positive but slices from the Tg12 PrP -/- genotype did not. Infectivity was present in 11 out of 11Tg 12 PrP+/-, whereas 0 out of 10 Tg12 PrP -/-. We successfully cultured POSCA brain slices infected with RML scrapie as confirmed by RT-QuIC and PK digestion assays. Biochemical studies revealed degenerative changes and oxidative damage in the prion infected slice cultures. Our results demonstrate that combining brain slice culture models of prion diseases with the RT-QuIC assay offers a promising platform for studying the mechanisms of prion proteinopathies as well as for screening anti-prion therapeutics.


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