NR AWKV
AU Nakamura,I.; Wu,G.Y.; Saeki,K.; Matsumoto,Y.; Toniolo,A.; Onodera,T.
TI Study of subcellar localization of PrPc using cell lines transfected with Prnp genes
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions CE-34
PT Konferenz-Poster
AB PrPc is found predominantly on the surface of neurons, attached by a glycoinositol phospholipids anchor, while PrPsc has been seen accumulating in intracellulary cytoplasmic vesicles. Previously we showed that PrP has been detected in the nuclei of uninfected cells. In this study PrPc was detected more in detail using Prnp -/transfected cell lines. Cell lines used in this study were, type 2 PrP cell line (HpL 3-4), and mouse or bovine PrP gene transfected cell lines, designated HpL3-4 mPrP or HpL3-4 bPrP. Type-1 PrP-/- cell lines (HpL2), and mouse or bovine PrP transfectant, NpL2 mPrP or NpL2bPrP cell lines were also used. Wild-type mouse neuronal cell line NB3-2 was used as a control. Anti-PrP monoclonal antibodies (mAbs) T2 and 1D12 were used to detect PrP. Fluorescence were observed with a fluorescence microscope and confocal microscope. T2 mAb detected immunoreactivity of PrP predominantly on the surface of HpL3-4 mPrP, HpL3-4 bPrP, NpL2mPrP, NpL2bPrP and NB3-2 cells. On the other hand 1D12 mAb detected immunoreactivity of PrP on the surface and nuclei of HpL3-4 bPrP, NpL2 mPrP and, NpL 2bPrP when fixed with acetone. NB3-2 and HpL3-4 mPrP did not show fluorescence in this condition. However several wild type neural cell lines showed intranuclear positive fluorescence using 1D12. T2 mAb showed reactivity to mouse and bovine PrP on the cell surface. 1D12 mAb showed reactivity to bovine PrP on the cell membrane and organelle inside the nuclei. Role of PrPc in the nuclei using NpL2mPrP could be studied during the course of infection using mouse adapted scrapie agent.
AD I. Nakamura, G.Y. Wu, K. Saeki, Y. Matsumoto, T. Onodera: School of Agriculture and Life Sciences, University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan; A. Toniolo: University of Insubria, viale Borri 57, Varese 2100, Italy. E-mail: aonoder@mail.ecc.u-tokyo.ac.jp
SP englisch
PO Italien