Uptake and neuritic transport of scrapie prion protein coincident with infection of neuronal cells.
Invasion of the nervous system and neuronal spread of infection are critical, but poorly understood, steps in the pathogenesis of transmissible spongiform encephalopathies or prion diseases. To characterize pathways for the uptake and intraneuronal trafficking of infectious, protease-resistant prion protein (PrP-res), fluorescent-labeled PrP-res was used to infect a neuronally derived murine cell line (SN56) and adult hamster cortical neurons in primary culture. Concurrent with the establishment of persistent scrapie infection, SN56 cells internalized PrP-res aggregates into vesicles positive for markers for late endosomes and/or lysosomes but not synaptic, early endocytic, or raft-derived vesicles. Internalized PrP-res was then transported along neurites to points of contact with other cells. Similar trafficking was observed with dextran, Alzheimer's Abeta1-42 fibrils and noninfectious recombinant PrP fibrils, suggesting that PrP-res is internalized by a relatively nonspecific pinocytosis or transcytosis mechanism. Hamster cortical neurons were also capable of internalizing and disseminating exogenous PrP-res. Similar trafficking of exogenous PrP-res by cortical neurons cultured from the brains of PrP knock-out mice showed that uptake and neuritic transport did not require the presence of endogenous cellular PrP. These experiments visualize and characterize the initial steps associated with prion infection and transport within neuronal cells.
Magalhães
Ana Cristina
AC
Laboratory of Persistent Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana 59840, USA.
Baron
Gerald S
GS
Lee
Kil Sun
KS
Steele-Mortimer
Olivia
O
Dorward
David
D
Prado
Marco A M
MA
Caughey
Byron
B
D000818:Animals; D001692:Biological Transport; D015153:Blotting, Western / Q000379:methods; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D002772:Cholera Toxin / Q000378:metabolism; D006224:Cricetinae; D003952:Diagnostic Imaging / Q000379:methods; D019286:Endopeptidase K / Q000494:pharmacology; D005455:Fluorescent Antibody Technique / Q000379:methods; D049452:Green Fluorescent Proteins / Q000378:metabolism; D051379:Mice; D016296:Mutagenesis / Q000502:physiology; D009422:Nervous System Diseases / Q000378:metabolism*; D016501:Neurites / Q000502:physiology* / Q000821:virology; D009474:Neurons / Q000378:metabolism* / Q000821:virology; D018620:PrPSc Proteins / Q000378:metabolism*; D021381:Protein Transport / Q000502:physiology; D013997:Time Factors; D014162:Transfection / Q000379:methods; D020691:rab GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism; D000090802:rab7 GTP-Binding Proteins
D018620:PrPSc Proteins; D000090802:rab7 GTP-Binding Proteins; D049452:Green Fluorescent Proteins; D002772:Cholera Toxin; D019286:Endopeptidase K; D020691:rab GTP-Binding Proteins