Abstract
The neuropathological diagnosis of Creutzfeldt‐Jakob disease relies on the immunohistochemical demonstration of the proteinase‐K resistant form of the prion protein (PrPres) in the brain tissue. The antigenicity of PrPres is strongly reduced by the formalin solution widely used to fix the tissue, thus the PrPres immunoreactivity is inconsistently detectable in formalin‐fixed tissue. A better PrPres immunostaining can be obtained by using Carnoy's fixing solution, which is composed of ethanol, chloroform and acetic acid (6:3:1). PrPres can easily be extracted from Carnoy's‐fixed, paraplast‐embedded tissue. Accordingly, Carnoy's‐fixed tissue can prior to immunolabeling be subjected to proteinase K and guanidine thio‐cyanate, which respectively eliminate the normal cellular form of prion protein and promote protein denaturation. In comparison with the best protocols for formalin‐fixed tissue (i.e. ‐ hydrolytic autoclaving or autoclaving in distilled water followed by formic acid and guanidine thiocyanate), PrPres immunostaining carried out on sections cut from Carnoy's‐fixed, paraplast‐embedded tissue blocks and subjected to proteinase K and guanidine thiocyanate, proved more successful to detect and map both diffuse and focal PrPres immunoreactivity, and to correlate the immunoreactivity pattern with MV polymorphism at PRNP codon 129 and PrPres banding and glycosyla‐tion pattern revealed by Western blot.
Full Text
The Full Text of this article is available as a PDF (677.3 KB).
References
- 1. Bell JE, Gentleman SM, Ironside JW, McCardle I, Lantos PI, Doey L, Lowe J, Fergusson J, Lutherst P, McQuaid S, Allen IV (1997) Prion protein immunohistochemistry ‐ UK five centres consensus report. Neuropathol Appl Neurobiol 23:26–35. [PubMed] [Google Scholar]
- 2. Brown P, Wolff A, Gajdusek CD (1990) A simple and effective method for inactivating virus infectivity in formalin‐fixed tissue samples from patients with Creutzfeldt‐Jakob disease. Neurology 40:887–890. [DOI] [PubMed] [Google Scholar]
- 3. Budka H, Aguzzi, A , Brown, P , Brucher J‐M, Bugiani O, Collinge J, Diringer H, Gullotta F, Haltia M, Hauw J‐J, Ironside JW, Kretzschmar HA, Lantos PL, Masullo C, Pocchiari M, Schlote W, Tateishi J, Will RG (1995) Tissue handling in suspected Creutzfeldt‐Jakob disease (CJD) and other human spongiform encephalopathies (prion diseases). Brain Pathol 5:319–322. [DOI] [PubMed] [Google Scholar]
- 4. Corti CJ, Larcher F, Chesner J, Aldaz CM (1988) Polyacrylamide gel electrophoresis and immunoblotting of proteins extracted from paraffin‐embedded tissue sections. J Histochem Cytochem 36:547–550. [DOI] [PubMed] [Google Scholar]
- 5. Kascsak RJ, Fersko R, Pulgiano R, Rubenstein R, Carp RI (1997) Immunodiagnosis of prion disease. Immunol Invest 26:259–268. [DOI] [PubMed] [Google Scholar]
- 6. Kitamoto T, Shrin R‐W, Doh‐ura K, Tomokane N, Miyazono M, Muramoto T, Tateishi J (1992) Abnormal iso‐form of prion protein accumulates in the synaptic structures of the central nervous system of patients with Creutzfeldt‐Jakob disease. Am J Pathol 140:1285–1294. [PMC free article] [PubMed] [Google Scholar]
- 7. Kocisko DA, Come JH, Priola SA, Chesebro B, Raymond GJ, Lansbury PT, Caughey B (1994) Cell‐free formation of protease‐resistant prion protein. Nature 370:471–474. [DOI] [PubMed] [Google Scholar]
- 8. Parchi P, Castellani R, Capellari S, Ghetti B, Young K, Chen SG, Farlow MR, Dickson DW, Sima AAF, Trojanowski JQ, Petersen RB, Gambetti P (1996) Molecular basis of phenotypic variability in sporadic Creutzfeldt‐Jakob disease. Ann Neurol 39:767–778. [DOI] [PubMed] [Google Scholar]
- 9. Puoti G, Giaccone G, Rossi G, Canciani B, Bugiani O, Tagliavini F (1999) Sporadic Creutzfeldt‐Jakob disease. Co‐occurrence of different types of PrPSc in single patients. Neurology, In Press. [DOI] [PubMed] [Google Scholar]
- 10. Taraboulos A, Jendroska K, Serban D, Yang S‐Y, DeArmond SJ, Prusiner SB (1992) Regional mapping of prion proteins in brain. Proc Natl Acad Sci USA 89:7620–7624. [DOI] [PMC free article] [PubMed] [Google Scholar]