Immunohistochemical detection of disease-associated prion protein in the intestine of cattle naturally affected with bovine spongiform encephalopathy by using an alkaline-based chemical antigen retrieval method.
An alkaline-based chemical antigen retrieval pretreatment step was used to enhance immunolabeling of disease-associated prion protein (PrP(Sc)) in formalin-fixed and paraffin-embedded tissue sections from cattle naturally affected with bovine spongiform encephalopathy (BSE). The modified chemical method used in this study amplified the PrP(Sc) signal by unmasking PrP(Sc) compared with the normal cellular prion protein. In addition, this method reduced nonspecific background immunolabeling that resulted from the destruction of the residual normal cellular form of prion protein, and reduced the treatment time compared with the usual autoclave pretreatment step. Immunolabeled PrP(Sc) was thereby clearly detected in the myenteric plexus of the ileum in naturally occurring BSE cattle.
Okada
Hiroyuki
H
Prion Disease Research Center, National Institute of Animal Health, Ibaraki, Japan. okadahi@affrc.go.jp
Iwamaru
Yoshihumi
Y
Imamura
Morikazu
M
Masujin
Kentaro
K
Yokoyama
Takashi
T
Mohri
Shirou
S
D000818:Animals; D000941:Antigens / Q000302:isolation & purification; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism / Q000473:pathology; D002417:Cattle; D002418:Cattle Diseases / Q000473:pathology*; D002432:Cecum / Q000473:pathology; D003106:Colon / Q000473:pathology; D016643:Encephalopathy, Bovine Spongiform / Q000175:diagnosis / Q000473:pathology* / Q000635:transmission; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007082:Ileum / Q000378:metabolism / Q000473:pathology; D007150:Immunohistochemistry / Q000379:methods / Q000662:veterinary; D007422:Intestines / Q000473:pathology*; D007583:Jejunum / Q000473:pathology; D051379:Mice; D010581:Peyer's Patches / Q000473:pathology; D018620:PrPSc Proteins / Q000378:metabolism; D011328:Prions / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D012007:Rectum / Q000473:pathology; D013116:Spinal Cord / Q000378:metabolism / Q000473:pathology; D013788:Thalamus / Q000473:pathology
D000941:Antigens; D018620:PrPSc Proteins; D011328:Prions