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Background  In prion disease, the peripheral expression of PrP C  is necessary for the transfer of infectivity to the central nervous system. The spleen is involved in neuroinvasion and neural dissemination in prion diseases but the nature of this involvement is not known. The present study undertook the investigation of the spatial relationship between sites of PrP Sc  accumulation, localisation of nerve fibres and PrP C  expression in the tissue compartments of the spleen of scrapie-inoculated and control sheep.    Methodology/Principal Findings  Laser microdissection and quantitative PCR were used to determine PrP mRNA levels and results were compared with immunohistochemical protocols to distinguish PrP C  and PrP Sc  in tissue compartments of the spleen. In sheep experimentally infected with scrapie, the major sites of accumulation of PrP Sc  in the spleen, namely the lymphoid nodules and the marginal zone, expressed low levels of PrP mRNA. Double immunohistochemical labelling for PrP Sc  and the pan-nerve fibre marker, PGP, was used to evaluate the density of innervation of splenic tissue compartments and the intimacy of association between PrP Sc  and nerves. Some nerve fibres were observed to accompany blood vessels into the PrP Sc -laden germinal centres. However, the close association between nerves and PrP Sc  was most apparent in the marginal zone. Other sites of close association were adjacent to the wall of the central artery of PALS and the outer rim of germinal centres.    Conclusions/Significance  The findings suggest that the degree of PrP Sc  accumulation does not depend on the expression level of PrP C . Though several splenic compartments may contribute to neuroinvasion, the marginal zone may play a central role in being the compartment with most apparent association between nerves and PrP Sc .

PrP Expression, PrPSc Accumulation and Innervation of Splenic Compartments in Sheep Experimentally Infected with Scrapie