Immunolocalisation of PrPSc in scrapie-infected N2a mouse neuroblastoma cells by light and electron microscopy

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2009
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Veith, Nathalie Monika
Schulz-Schaeffer, Walter J.
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European Journal of Cell Biology. 2009, 88(1), pp. 45-63. ISSN 0171-9335. eISSN 1618-1298. Available under: doi: 10.1016/j.ejcb.2008.08.001
Zusammenfassung

The causative agent of transmissible spongiform encephalopathies (TSE) is PrPSc, an infectious, misfolded isoform of the cellular prion protein (PrPC). The localisation and trafficking of PrPSc and sites of conversion from PrPC to PrPSc are under debate, particularly since most published work did not discriminate between PrPC and PrPSc. Here we describe the localisation of PrPC and PrPSc in a scrapie-infected neuroblastoma cell line, ScN2a, by light and electron microscopic immunolocalisation. After eliminating PrPC with proteinase K, PrPSc was detected at the plasma membrane, endocytosed via clathrin-coated pits and delivered to early endosomes. Finally, PrPSc was detected in late endosomes/lysosomes. As we detected PrPSc at the cell surface, in early endosomes and in late endosomes/lysosomes, i.e. locations where PrPC is also present, our data imply that the conversion process could take place at the plasma membrane and/or along the endocytic pathway. Finally, we observed the release of PrPC/PrPSc via exocytotic pathways, i.e. via exosomes and as an opaque electron-dense mass which may represent a mechanism of intercellular spreading of infectious prions.

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Fachgebiet (DDC)
570 Biowissenschaften, Biologie
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Electron microscopy, Immunofluorescence, Early endosomes, Late endosomes, Lysosomes, Clathrincoated pits, Exosomes
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ISO 690VEITH, Nathalie Monika, Helmut PLATTNER, Claudia STÜRMER, Walter J. SCHULZ-SCHAEFFER, Alexander BÜRKLE, 2009. Immunolocalisation of PrPSc in scrapie-infected N2a mouse neuroblastoma cells by light and electron microscopy. In: European Journal of Cell Biology. 2009, 88(1), pp. 45-63. ISSN 0171-9335. eISSN 1618-1298. Available under: doi: 10.1016/j.ejcb.2008.08.001
BibTex
@article{Veith2009Immun-8445,
  year={2009},
  doi={10.1016/j.ejcb.2008.08.001},
  title={Immunolocalisation of PrPSc in scrapie-infected N2a mouse neuroblastoma cells by light and electron microscopy},
  number={1},
  volume={88},
  issn={0171-9335},
  journal={European Journal of Cell Biology},
  pages={45--63},
  author={Veith, Nathalie Monika and Plattner, Helmut and Stürmer, Claudia and Schulz-Schaeffer, Walter J. and Bürkle, Alexander}
}
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