NR AXTS
AU Nouvel,V.; Picoli,C.; Bourai,M.; Aubry,F.; Lenuzza,N.; Charveriat,M.; Correia,E.; Reboul,M.; Deslys,J.P.; Mouthon,F.
TI Investigating the Relationship Between Cell and Prion
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Pathology and Pathogenesis P03.116
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
At the cellular level, the molecular dysfunctions caused by Prion infection are poorly understood. Several genes and proteins have been proposed to be abnormally regulated after prion infection without any unified concept of cellular reaction. We have characterized another cellular reaction based on different PrPc levels between infected and uninfected cells.
To understand these changes, we focused on the modification of PrP expression at the transcriptional level and at the membrane level. During Prion infection the level of PrPc is generally considered to be constant throughout the disease process. However, an over-expression of PrPc has been described during neuronal differentiation via a MAPK pathway activation in the PC12 system.
The aim of our work was to study Prnp promoter activity during infection and differentiation. For this purpose, we used a reporter vector encoding Green Fluorescent Protein under the control of the whole murine prion promoter, transfected in different murine-susceptible neuronal cell lines (SN56 and GT1).To focus on membrane PrP modifications during infection, we used two different approaches: on the one hand, the use of a functional exogenous whole PrP (containing the GPI anchor and 3F4 tagged) and, on the other hand, the use of plasmids encoding a GFP-PrP and YFP-GPI. Here, we present different methods (flow cytometry, western blotting, realtime PCR, immunocytochemistry) to elucidate this type of cellular reaction. These complementary approaches should allow us to characterize cellular reaction specifically linked to PrPres accumulation, and may possibly lead to the concept of innate immunity of cells to Prion infection.
AD V. Nouvel, C. Picoli, M. Bourai, N. Lenuzza, M. Charveriat, E. Correia, M. Reboul, J.P. Deslys, F. Mouthon, CEA/DSV/IMETI/SEPIA, France; F. Aubry, Alliance Biosecure Foundation, France
SP englisch
PO Schottland
EA pdf-Datei und Poster (Posterautoren ergänzt um L. Bellanger)