Prion Protein (PrPc) Immunocytochemistry and Expression of the Green Fluorescent Protein Reporter Gene under Control of the Bovine PrP Gene Promoter in the Mouse Brain

Journal of Comparative Neurology

Volumn 473, Issue 2, 2004, Pages 244-269

  Bailly, Yannick ^a,e   Haeberlé, Anne Marie ^a   Blanquet Grossard, Françoise ^b,d   Chasserot Golaz, Sylvette ^a   Grant, Nancy ^a   Schulze, Tobias ^b   Bombarde, Guy ^a   Grassi, Jacques ^c   Cesbron, Jean Yves ^b,d   Lemaire Vieille, Catherine ^b,d  

^a Centre de Neurochimie   (France)

^b INSTITUT PASTEUR DE LILLE   (France)

^c Service de Pharmacologie et Immunoanalyse   (France)

^d UNIV GRENOBLE ALPES   (France)

^e CENTRE DE NEUROCHIMIE   (France)

Author keywords

Central nervous system; Electron microscopy; Histofluoreseence; Immunogold; Transgenic mouse

Indexed keywords

GREEN FLUORESCENT PROTEIN; PRION PROTEIN; PHOTOPROTEIN;

ANIMAL TISSUE; ARTICLE; BRAIN REGION; CELL ADHESION; CELL ORGANELLE; CELL SURFACE; CELL TYPE; CENTRAL NERVOUS SYSTEM; CONTROLLED STUDY; GENE CONTROL; GENE REPLICATION; GENETIC TRANSCRIPTION; HIPPOCAMPUS; HOST CELL; IMMUNOCYTOCHEMISTRY; IMMUNODETECTION; IMMUNOGOLD LABELING; MOUSE; NEOCORTEX; NEUROPATHOLOGY; NONHUMAN; OLFACTORY BULB; PRION DISEASE; PRIORITY JOURNAL; PROMOTER REGION; PROTEIN EXPRESSION; PROTEIN LOCALIZATION; PUTAMEN; REPORTER GENE; TARGET CELL; THALAMUS; TRANSGENIC MOUSE; ANIMAL; BIOSYNTHESIS; BRAIN; CATTLE; COMPARATIVE STUDY; GENE EXPRESSION REGULATION; GENETICS; IMMUNOHISTOCHEMISTRY; METABOLISM; PHYSIOLOGY; ULTRASTRUCTURE;

ANIMALS; BRAIN; CATTLE; GENE EXPRESSION REGULATION; GENES, REPORTER; GREEN FLUORESCENT PROTEINS; IMMUNOHISTOCHEMISTRY; LUMINESCENT PROTEINS; MICE; MICE, TRANSGENIC; PRPC PROTEINS;

EID: 2342472104     PISSN: 00219967     EISSN: None     Source Type: Journal    
DOI: 10.1002/cne.20117     Document Type: Article

References (97)

1. Sc) into contiguous membranes. EMBO J 21:10:31-1040.
2. Basler K, Oesch B, Scott M, Westaway D, Wächli M, Groth DF, McKinley MP, Prusiner S, Weissmann C. 1986. Scrapie and cellular Prp isoforms are encoded by the same chromosomal gene. Cell 46:417-428.
3. Belichenko PV, Brown D, Jeffrey M, Fraser JR. 2000. Dendritic and synaptic alterations of hippocampal pyramidal neurons in scrapie-infected mice. Neuropathol Appl Neurobiol 26:143-149.
4. Bendheim PE, Brown HR, Rudelli RD, Scala LJ, Goller NL, Wen GY, Kascsak RJ, Cashman NR, Bolton DC. 1992. Nearly ubiquitous tissue distribution of the scrapie agent precursor protein. Neurology 42:149-156.
5. Bolton DC, McKinley MP, Prusiner SB. 1982. Identification of a protein that purifies with the scrapie prion. Science 218:1309-1311.
6. Borchelt DR, Taraboulos A, Prusiner SB. 1992. Evidence for synthesis of scrapie prion proteins in the endocytotic pathway. J Biol Chem 267: 16188-16199.
7. Bouzamondo E, Milroy AM, Ralston HJ, Prusiner SB, De Armond SJ. 2000. Selective neuronal vulnerability during experimental scrapie infection: insights from an ultrastructural investigation. Brain Res 874:210-215.
8. Brandner S, Isenmann S, Raeber A, Fischer M, Sailer A, Kobayashi Y, Marino S, Weissmann C, Aguzzi A. 1996. Normal host prion protein necessary for scrapie-induced neurotoxicity. Nature 379:339-343.
9. Brown D, Belichenko P, Sales J, Jeffrey M, Fraser JR. 2001. Early loss of dendritic spines in murine scrapie revealed by confocal analysis. Neuroreport 12:179-183.
10. Brown DR. 2001. Prion and prejudice: normal protein and the synapse. Trends Neurosci 24:85-90.
11. Brown HR, Goller NL, Rudelli RD, Merz GS, Wolfe GC, Wisniewsky HM, Robakis NK. 1990. The mRNA encoding the scrapie agent protein is present in a variety of non-neuronal cells. Acta Neuropathol 80:1-6.
12. Bruce ME, McBride PA, Farquhar CF. 1989. Precise targeting of the pathology of the sialoglycoprotein, PrP, and vacuolar degeneration in mouse scrapie. Neurosci Lett 102:1-6.
13. Bruce ME, McBride PA, Jeffrey M, Scott JR. 1994. PrP in pathology and pathogenesis in scrapie-infected mice. Mol Neurobiol 8:105-112.
14. Büeler H, Fischer M, Lang Y, Bluetmann H, Lip H, De Armond SJ, Prusiner SB, Aguet M, Weissmann C. 1992. Normal development and behavior of mice lacking the neuronal cell-surface PrP protein. Nature 356:577-582.
15. Büeler H, Aguzzi A, Sailer A, Greiner RA, Autenried P, Aguet M, Weissmann C. 1993. Mice devoid of PrP are resistant to scrapie. Cell 73: 1339-1347.
16. Cagampang FRA, Whatley SA, Mitchell AL, Powell JF, Campbell IC, Coen CW. 1999. Circadian regulation of prion protein messenger mRNA in the rat forebrain: a widespread and synchronous rhythm. Neuroscience 91:1201-1204.
17. Caughey B, Race RE, Chesebro B. 1988. Detection of prion protein mRNA in normal and scrapie-infected tissue and cell lines. J Gen Virol 69: 711-716.
18. Chen SG, Teplow DB, Parchi P, Teller JK, Gambetti P, Autilio-Gambetti L. 1995. Truncated forms of the human prion protein in normal brain and prion diseases. J Biol Chem 270:19173-19180.
19. Collinge J, Whittington MA, Sidle KCL, Smith CJ, Palmer MS, Clarke AR, Jefferys JGR. 1994. Prion protein is necessary for normal synaptic function. Nature 370:295-297.
20. Cunningham C, Deacon R, Wells H, Boche D, Waters S, Picanco Diniz C, Scott H, Rawlins JNP, Perry VH. 2003. Synaptic changes characterize early behavioral signs in the ME7 model of murine prion disease. Eur J Neurosci 17:2147-2155.
21. Daniels M, Brown DR. 2002. Purification and preparation of prion protein: synaptic superoxide dismutase. Methods Enzymol 349:258-267.
22. De Armond SJ, Mobley WC, DeMott DL, Barry RA, Beckstead JH, Prusiner SB. 1987. Changes in the localization of brain prion proteins during scrapie infection. Neurology 37:1271-1280.
23. De Armond SJ, Yang SL, Lee A, Bowler R, Taraboulos A, Groth D, Prusiner SB. 1993. Three scrapie prion isolates exhibit different accumulation patterns of the prion protein scrapie isoform. Proc Natl Acad Sci U S A 90:6449-6453.
24. De Armond SJ, Sanchez H, Yehiely F, Qiu Y, Ninchak-Casey A, Dagget V, Camerino AP, Cayetano J, Rogers M, Groth D, Torchia M, Tremblay P, Scott MR, Cohen FE, Prusiner SB. 1997. Selective neuronal targeting in prion disease. Neuron 19:1337-1348.
25. c glycoform hetorogeneity as a function of brain region: implications for selective targeting of neurons by prion srains. J Neuropathol Exp Neurol 58:1000-1009.
26. Demart S, Fournier J-G, Creminon C, Frobert Y, Lamoury F, Marce D, Lasmézas C, Dormont D, Grassi J, Deslys J-P. 1999. New insight into abnormal prion protein usig monoclonal antibodies. Biochem Biophys Res Commun 265:652-657.
27. Diez M, Koistinaho J, De Armond SJ, Groth D, Prusiner SB, Hökfelt T. 1997. Marked decrease of neuropeptide Y Y2 receptor binding sites in the hippocampus in murine prion disease. Proc Natl Acad Sci U S A 94:13267-13272.
28. Feng G, Mellor RH, Bernstein M, Keller-Peck C, Nguyen QT, Wallace M, Nerbonne JM, Lichtman JW, Sanes JR. 2000. Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP. Neuron 28:41-51.
29. Ferrer I, Puig B, Blanco R, Marti E. 2000. Prion protein deposition and abnormal synaptic protein expression in the cerebellum in Creutzfeldt-Jakob disease. Neuroscience 97:715-726.
30. Follet J, Lemaire-Vieille C, Blanquet-Grossard F, Podevin-Dimster V, Lehmann S, Chauvin JP, Decavel JP, Varea R, Grassi J, Fontes M, Cesbron JY. 2002. PrP expression and replication by Schwann cells: implications in prion spreading. J Virol 76:2434-2439.
31. Ford MJ, Burton LJ, Li H, Graham CH, Frobert Y, Grassi J, Hall SM, Morris RJ. 2002. A marked disparity between the expression of prion protein and its message by neurones of the CNS. Neuroscience 111: 533-551.
32. Fournier JG, Grigoriev V. 2001. Prion diseases: contribution of high-resolution immunomorphology. J Cell Mol Med 5:367-377.
33. c) in synaptic boutons of normal hamster hippocampus. CR Acad Sci 318:339-344.
34. Fournier JG, Escaig-Haye F, Grigoriev V. 2000. Ultrastructural localization of prion proteins: physiological and pathological implications. Microsc Res Techniq 50:76-88.
35. Fraser H, Dickinson AG. 1973. Scrapie in mice: agent strain differences in the distribution and intensity of gray matter vacuolation. J Comp Pathol 83:29-40.
36. c expression in the peripheral nervous system is a determinant of prion neuroinvasion. J Gen Virol 81:2813-2821.
37. Grigoriev V, Escaig-Haye F, Streichenberger N, Kopp N, Langeveld J, Brown P, Fournier JG. 1999. Submicroscopic immunomorphology of PrPsc in a new-variant CJD case. Neurosci Lett 264:57-60.
38. Guentchev M, Groschup MR, Kodek R, Liberski PP, Budka H. 1998. Severe, early and selective loss of a subpopulation of GABAergic inhibitory neurons in experimental transmissible spongiform encephalopathies. Brain Pathol 8:615-623.
39. Haeberlé A-M, Ribaut-Barassin C, Bombarde G, Mariani J, Hunsmann G, Grassi J, Bailly Y. 2000. Synaptic prion protein immunoreactivity in the rodent cerebellum. Microsc Res Techniq 75:50-66.
40. Harris DA, Lele P, Snider WD. 1993a. Localization of the mRNA for a chicken prion protein by in situ hybridization. Proc Natl Acad Sci U S A 90:4309-4313.
41. Harris DA, Huber MT, van Dijken P, Shyng SL, Chait BT, Wang R. 1993b. Processing of a cellular prion protein: identification of N- and C-terminal cleavage sites. Biochemistry 32:1009-1016.
42. Hecker R, Taraboulos A, Scott M, Pan KM, Yang SL, Torchia M, Jendroska K, De Armond SJ, Prusiner SB. 1992. Replication of distinct scrapie isolates is region specific in brain of transgenic mice and hamster. Genes Dev 6:1213-1228.
43. Heggebø R, Gonzalez L, Press CM, Gunnes G, Espenes A, Jeffrey M. 2003. Disease-associated PrP in the enteric nervous system of scrapie-affected Suffolk sheep. J Gen Virol 84:1327-1338.
44. Herms J, Tings T, Gali S, Madlung A, Giese A, Siebert H, Schürmann P, Windl O, Brose N, Kretzschmar H. 1999. Evidence of presynaptic location and function of the prion protein. J Neurosci 19:8866-8875.
45. Hsu SM, Raine L, Fanger H. 1981. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques. J Histochem Cytochem 29:77-580.
46. Jansen GH, Vogelaar CF, Elshof SM. 2001. Distribution of cellular prion protein in normal cerebral cortex-does it have relevance to Creutzfeldt-Jakob disease? Clin Chem Lab Med 39:294-298.
47. Jefferys JGR, Empson RM, Wittington MA, Prusiner SB. 1994. Scrapie infection of transgenic mice leads to network and intrinsic dysfunction of cortical and hippocampal neurons. Neurobiol Dis 1:25-30.
48. Jeffrey M, Goodsir CM, Holliman A, Higgins RJ, Bruce ME, McBride PA, Fraser JR. 1998. Determination of the frequency and distribution of vascular and parenchymal amyloid with polyclonal and N-terminal-specific PrP antibodies in scrapie-affected sheep and mice. Vet Rec 142:534-537.
49. Jeffrey M, Halliday WG, Bell J, Johnston AR, McLeod, NK, Ingham C, Sayers AR, Brown DA, Fraser JR. 2000. Synapse loss associated with abnormal PrP precedes neuronal degeneration in the scrapie-infected murine hippocampus. Neuropathol Appl Neurobiol 26:41-54.
50. Jendroska K, Heinzel FP, Torchia M, Stowring L, Kretzschmar HA, Kon A, Stern A, Prusiner SB, De Armond SJ. 1991. Proteinase-resistant prion protein accumulation in Syrian hamster brain correlates with regional pathology and scrapie infectivity. Neurology 41:1482-1490.
51. Jimenez-Huete A, Lievens PMJ, Vidal R, Piccardo P, Ghetti B, Tagliavini F, Frangione B, Prelli F. 1998. Endogenous proteolytic cleavage of normal and disease-associated isoforms of the human prion protein in neural and non-neural tissues. Am J Pathol 153:1561-1572.
52. Kaneko K, Zulianello L, Scott IM, Cooper CM, Wallace AC, James TL, Cohen FE, Prusiner SB. 1997. Evidence for protein X binding to a discontinuous epitope on the cellular prion protein during scrapie prion propagation. Proc Natl Acad Sci U S A 94:10069-10074.
53. Kanu N, Imokawa Y, Drechsel DN, Williamson RA, Birkett CR, Bostock CJ, Brockes JP. 2002. Transfert of scrapie prion infectivity by cell contact in culture. Curr Biol 12:523-530.
54. Kitamoto T, Shin RW, Doh-ura K, Tomokane N, Miyazono M, Muramoto T, Tateishi J. 1992. Abnormal isoform of prion proteins accumulates in the synaptic structures of the central nervous system in patients with Creutzfeldt-Jakob disease. Am J Pathol 140:1285-1294.
55. Kociscko 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.
56. Koperek O, Kovacs GG, Ritchie D, Ironside JW, Budka H, Wick G. 2002. Disease-associated prion protein in vessel walls. Am J Pathol 161: 1979-1984.
57. Krasemann S, Groschup MH, Harmeyer S, Hunsmann G, Bodemer W. 1996a. Generation of monoclonal antibodies against human prion proteins in PrPo/o mice. Mol Med 2:725-734.
58. Krasemann S, Groschup MH, Hunsmann G, Bodemer W. 1996b. Induction of antibodies against human prion proteins (PrP) by DNA-mediated immunization of PrPo/o mice. J Immunol Methods 199:109-118.
59. Kretzschmar HA, Prusiner SB, Stowring LE, De Armond SJ. 1986. Scrapie prion proteins are synthesized in neurons. Am J Pathol 122:1-5.
60. Lainé J, Marc M-E, Sy M-S, Axelrad H. 2001. Cellular and subcellular morphological localization of normal prion protein in rodent cerebellum. Eur J Neurosci 14:47-56.
61. Lasmézas CI, Deslys JP, Demaimay R, Adjou KT, Haw JJ, Dormont D. 1996. Strain specific and common pathogenic events in murine models of scrapie and bovine spongiform encephalopathy. J Gen Virol 77:1601-1609.
62. Lehman A. 1974. Atlas stéréotaxique du cerveau de la souris. Paris: CNRS Ed.
63. Lemaire-Vieille C, Schulze T, Poidevin-Dimster V, Follet J, Bailly Y, Blanquet-Grossard F, Decavel J-P, Heinen E, Cesbron J-Y. 2000. Epithelial and endothelial expression of the green fluorescent protein reporter gene under the control of bovine prp regulatory sequence in transgenic mice. Proc Natl Acad Sci U S A 97:5422-5427.
64. Liu T, Zwingman T, Li R, Pan T, Wong BS, Petersen RB, Gambetti P, Herrup K, Sy MS. 2001. Differential expression of cellular prion protein in mouse brain as detected with multiple anti-PrP monoclonal antibodies. Brain Res 896:118-129.
65. Madore N, Smith KL, Graham CH, Jen A, Brady K, Hall S, Morris R. 1999. Functionally different GPI proteins are organized in different domains on the neuronal surface. EMBO J 18:6917-6926.
66. Maignien T, Lasmezas CI, Beringue V, Dormont D, Deslys JP. 1999. Pathogenesis of the oral route of infection in mice with scrapie and bovine spongiform encephalopathy agents. J Gen Virol 80:3035-3042.
67. Manson JC, McBride P, Hope J. 1992a. Expression of the PrP gene in the brain of Sinc congenic mice and its relationship to the development of scrapie. Neurodegeneration 1:45-52.
68. Manson JC, West JD, Thomson V, McBride P, Kaufinan MH, Hope J. 1992b. The prion protein gene: a role in mouse embryogenesis? Development 115:117-122.
69. Martins VR, Linden R, Prado MAM, Walz R, Sakamoto AC, Izquierdo I, Brentani RR. 2002. Cellular prion protein: on the road for functions. FEBS Lett 512:25-28.
70. McLennan NF, Rennisson KA, Bell JE, Ironside JW. 2001. In situ hybridisation analysis of PrP mRNA in human CNS tissues. Neuropathol Appl Neurobiol 27:373-383.
71. McMahon HEM, Mangé A, Nishida N, Créminon C, Casanava D, Lehmann S. 2001. Cleavage of the amino terminus of the prion protein by reactive oxygen species. J Biol Chem 276:2286-2291.
72. Mironov A, Latawiec D, Wille H, Bouzamondo-Bernstein E, Legname G, Williamson RH, Burton D, De Armond SJ, Prusiner SB, Peters PJ. 2003. Cytosolic prion protein in neurons. J Neurosci 23:7183-7193.
73. Moser M, Colello RJ, Pott U, Oesch B. 1995. Developmental expression of the prion protein gene in glial cells. Neuron 14:509-517.
74. Moya K, Salès N, Hässig R, Creminon C, Grassi J, Di Giambernardino L. 2000. Immunolocalization of the cellular prion protein in normal brain. Microsc Res Techniq 50:58-65.
75. Piccardo P, Safar J, Ceroni M, Gajdusek DC, Gibbs CJ. 1990. Immunohistochemical localization of prion protein in spongiform encephalopathies and normal brain tissue. Neurology 40:518-522.
76. Prinz M, Huber G, Macpherson AJ, Heppner FL, Glatzel M, Eugste HP, Wagner N, Aguzzi A. 2003. Oral prion infection requires normal numbers of Peyer's patches but not of enteric lymphocytes. Am J Pathol 162:1003-1011.
77. Prusiner SB. 1998. Prion protein biology. Cell 93:337-348.
78. Prusiner SB, Scott M, Foster D, Pan KM, Groth DF, Mirenda CA, Torchia M, Yang SL, Serban D, Carlson GA, Hoppe PC, Westaway D, De Armond SJ. 1990. Transgenic studies implicate interactions between homologous PrP isoforms in scrapie prion replication. Cell 63:673-686.
79. Race R, Oldstone M, Chesebro B. 2000. Entry versus blockade of brain infection following oral or intraperitoneal scrapie administration: role of prion protein expression in peripheral nerves and spleen. J Virol 74:828-833.
80. Raeber AJ, Race RE, Brandner S, Priola SA, Sailer A, Bessen RA, Mucke L, Manson J, Aguzzi A, Oldstone MBA, Weissmann C, Chesebro B. 1997. Astrocyte-specific expression of hamster prion protein (PrP) renders PrP knockout mice susceptible to hamster scrapie. EMBO J 20: 6057-6065.
81. Rodolfo K, Hassig R, Moya KL, Frobert Y, Grassi J, Di Giamberardino L. 1999. A novel cellular prion protein isoform present in rapid anterograde axonal transport. Neuroreport 10:3639-3644.
82. Sakaguchi S, Katamine S, Shigematsu K, Nakatani A, Moriuchi R, Nishida N, Kurokawa K, Nakaoke R, Sato H, Jishage K. 1995. Accumulation of proteinase K-resistant prion protein (PrP) is restricted by the expression level of normal PrP in mice inoculated with a mouse-adapted strain of the Creutzfeldt-Jakob disease agent. J Virol 69:7586-7592.
83. Sakagachi S, Katamine S, Nishida N, Moriuchi R, Shigematsu K, Sugimoto T, Nakatani A, Kataoka Y, Houtani T, Shirabe S, Okada H, Hasegawa S, Miyamoto T, Noda T. 1996. Loss of cerebellar Purkinje cells in aged mice homozygous for a disrupted PrP gene. Nature 380: 528-531.
84. Salès N, Rodolfo K, Hässig R, Faucheux B, DiGiamberardino L, Moya KL. 1998. Cellular prion protein localization in rodent and primate brain. Eur J Neurosci 10:2464-2471.
85. Salès N, Hässig R, Rodolfo K, DiGiamberardino L, Traiffort E, Ruat M, Frétier P, Moya KL. 2002. Developmental expression of the cellular prion protein in elongating axons. Eur J Neurosci 15:1163-1177.
86. Shupliakov O, Bloom O, Gustafsson JS, Kjaerulff O, Löw P, Tomilin N, Pieribone VA, Greengard P, Brodin L. 2002. Impaired recycling of synaptic vesicles after acute perturbation of the presynaptic actin cytoskeleton. Proc Natl Acad Sci U S A 99:14476-14481.
87. Siso S, Puig B, Varea R, Vidal E, Acin C, Prinz M, Montrasio F, Badiola J, Aguzzi A, Pumarola M, Ferrer I. 2002. Abnormal synaptic protein expression and cell death in murine scrapie. Acta Neuropathol 103: 615-626.
88. Stahl N, Borchelt DR, Hsiao K, Prusiner SB. 1987. Scrapie prion protein contains a phosphatidylinositol glycolipid. Cell 51:229-240.
89. c mRNA by in situ hybridization in brain, placenta, uterus and testis of rats. Intervirology 38:309-315.
90. Taraboulos A, Jendroska K, Serban D, Yang SL, De Armond SJ, Prusiner SB. 1992. Regional mapping of prion protein in brain. Proc Natl Acad Sci U S A 89:7620-7624.
91. Telling GC, Scott M, Mastrianni J, Gabizon R, Torchia M, Cohen FE, De Armond SJ, Prusiner SB. 1995. Prion propagation in mice expressing human and chimeric PrP transgenes implicates the interaction of cellular PrP with another protein. Cell 83:79-90.
92. Thackray AM, Klein MA, Bujdoso R. 2003. Subclinical prion disease induced by oral inoculation. J Virol 77:7991-7998.
93. Tomomura M, Rice DS, Morgan JI, Yuszaki M. 2001. Purification of Purkinje cells by fluorescence-activated cell sorting from transgenic mice that express green fluorescent protein. Eur J Neurosci 14:57-63.
94. van Keulen LJ, Vromans ME, van Zijderveld FG. 2002. Early and late pathogenesis of natural scrapie infection in sheep. APMIS 110:23-32.
95. Verghese-Nikolakaki S, Michaloudi H, Polymenidou M, Groschup MH, Papadopoulos GC, Sklaviadis T. 1999. Expression of the prion protein prion in the rat forebrain-an immunohistochemical study. Neurosci Lett 272:9-12.
96. Vorberg I, Priola SA. 2002. Molecular basis of scrapie strain glycoform variation. J Biol Chem 277:36775-36781.
97. Zanusso G, Ferrari S, Cardone F, Zampieri P, Gelati M, Fiorini M, Farinazzo A, Gardiman M, Cavallaro T, Bentivoglio M, Righetti PG, Pocchiari M, Rizzato N, Monaco S. 2003. Detection of pathologic prion protein in the olfactory epithelium in sporadic Creutzfeldt-Jakob disease. N Engl J Med 348:711-719.