title: Immunogold electron microscopy recognizes prion protein-associated particles prepared from scrapie-infected mouse brain.
abstract: Previous studies have proposed that the disease isoform of prion protein (PrPSc) is particulate. Our purpose was to search by electron microscopy (EM) for particles in fractions of density gradients prepared from differentially centrifuged homogenates of scrapie-infected, normal, and null mouse brain. Only mild detergents were used during the separation process. The low-density fractions derived from scrapie-infected brain were rich in PrP. Three morphologically distinct types of particle were observed. Type 1 particles, measuring approximately 6.8 nm in mean diameter, were found in abundance in the fractions of scrapie-infected brain at the peak PrP concentrations. They were often clumped and adherent to raft-like structures. Type 2 particles, in low-density fractions from normal brain, were similar to type 1 but were smaller, with the mean diameter measuring approximately 5.3 nm. Type 3 particles from null brain differed morphologically from types 1 and 2 and were not clumped. The low density of the particles indicated a lipid component, which was confirmed by lipid analysis. Immunogold EM using Mab 6H4 labeled a portion of the particles from scrapie mouse brain, but not those from normal or null brain. Dimensions of PrP suggest that the labeled particles carry a PrPSc dimer per particle.
authors.affiliation: Section of Academic Virology at St. Barts, Royal London Hospital and QMW Medical College, London, United Kingdom. dourmashkin@blueyonder.co.uk
authors.forename: Robert R
authors.initials: RR
authors.lastname: Dourmashkin
authors.forename: John S
authors.initials: JS
authors.lastname: Oxford
authors.forename: Lynne
authors.initials: L
authors.lastname: Bountiff
mesh_terms: D000818:Animals; D015153:Blotting, Western; D001923:Brain Chemistry; D002499:Centrifugation, Density Gradient / Q000379:methods; D005260:Female; D007150:Immunohistochemistry / Q000379:methods; D008055:Lipids / Q000032:analysis; D008297:Male; D051379:Mice; D008854:Microscopy, Electron / Q000379:methods; D008861:Microsomes; D018620:PrPSc Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000648:ultrastructure*; D012608:Scrapie / Q000473:pathology
chemical_list: D008055:Lipids; D018620:PrPSc Proteins