1.Dynamic analyses of PrP and PrP(Sc) in brain tissues of golden hamsters infected with scrapie strain 263K revealed various PrP forms.
Jian-Mei GAO ; Chen GAO ; Jun HAN ; Xiao-Bo ZHOU ; Xin-Li XIAO ; Jin ZHANG ; Lan CHEN ; Bao-Yun ZHANG ; Tao HONG ; Xiao-Ping DONG
Biomedical and Environmental Sciences 2004;17(1):8-20
OBJECTIVETo expatiate dynamic changes in hamsters infected with scrapie strain 263K, to observe the presence and aggravation of various forms of PrP and PrP(Sc) during incubation period, and to probe primarily the relationship between the onset of clinic manifestations and the presence of different PrP(Sc) forms.
METHODSHamster-adapted scrapie strain 263K was intracerebrally inoculated into hamsters. Different forms of PrP and PrP(Sc) were monitored dynamically by Western blot and immuno-histochemical assays. The presence of scrapie-associated fibril (SAF) was assayed by electron microscopy analysis (EM) and immuno-golden EM.
RESULTSPrP(Sc) was initially detected in the brain tissues of the animals in 20 days post-inoculation by immunohistochemistry and 40 days with Western blot. Quantitative evaluations revealed that the amounts of PrP and PrP(Sc) in brain tissues increased along with the incubation. Several high and low molecular masses of PrP were seen in the brains of the long-life span infected animals. Deglycosylation assays identified that the truncated PrP in the infected brains showed similar glycosylation patterns as the full-length PrP. The presence of short fragments was seemed to relate with the onset of clinical conditions.
CONCLUSIONThese results indicate that infectious agents exist and accumulate in central nerve system prior to the onset of the illness. Various molecular patterns of PrP(Sc) may indwell in brain tissues during the infection.
Animals ; Blotting, Western ; Brain ; metabolism ; ultrastructure ; Cricetinae ; Disease Models, Animal ; Female ; Glycosylation ; Immunohistochemistry ; Mesocricetus ; Microscopy, Electron ; PrP 27-30 Protein ; analysis ; metabolism ; PrPC Proteins ; analysis ; metabolism ; PrPSc Proteins ; analysis ; metabolism ; Scrapie ; metabolism ; pathology