NR AWEU
AU Ferrari,S.; Fasoli,E.; Zanusso,G.; Gelati,M.; Ghetti,B.; Monaco,S.
TI Immunocytochemistry and immunoelectron microscopy of PrP amyloid plaques in human prion disorders
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions PA-15
PT Konferenz-Poster
AB In human prion disorders, few studies have been addressed to immunoelectron microscopic investigations of prion protein-associated amyloid plaques, due to the lack of anti-PrP antibodies able to recognize pathological PrP in glutaraldehyde-fixed specimens. We studied cerebellar sections from patients deceased from sporadic and familial human prion disorders characterized by amyloid plaques. Selected paraffin blocks, containing PrP amyloid plaques were deparaffinizzated, fixed in glutaraldehyde and embedded in Epon. PrP immunogold staining was obtained on semithin and ultrathin sections etched with potassium methoxide or sodium periodate, and treated with formic acid. By the use of different antibodies we compared the pattern of PrP deposition on paraffin sections with PrP staining in epoxy semithin and ultrathin sections. Among several monoclonal anti-PrP antibodies we selected SA65 (previously obtained in our laboratory) that stained PrP in etched thin sections treated with formic acid. In individual case, paraffin and epoxy thin sections stained with SA65 showed a similar pattern of PrP deposition. In immunoelectron microscopy SA65 selectively decorated amyloid fibrils without relevant background of gold particles. The PrP immunogold staining of amyloid plaques showed significant differences between genetic and sporadic human prion disorders. These data confirm the validity of monoclonal antibody SA65 in PrP staining by the use of post-embedding immunoelectron microscopy methods. We describe the variety of patterns of PrP deposition in amyloid plaques of different human prion disorders. IEM methods complement immunocytochemical and biochemical studies of prion disorders and can help in understanding prion biology
AD S. Ferrari, E. Fasoli,GT. Zanusso, M. Gelati, S. Monaco: Department of Neurological and Visual Sciences, Section of Clinical Neurology, University of Verona, Italy; B. Ghetti: Department of Pathology and Laboratory Medicine and Indiana Alzheimer Disease Center, Indiana University School of Medicine, Indianapolis, IN, USA. E-mail: gianluigi.zanusso@univr.it
SP englisch
PO Italien