NR ATIL
AU Kranich,J.; Miele,G.; Seeger,H.; Aguzzi,A.
TI Transcriptional and proteomic consequences of inhibition of prion protein expression in neuroblastoma cells
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Cell Biology of PrPc and PrPsc CELL-43
PT Konferenz-Poster
AB
Despite intensive efforts, the physiological function of PrP still remains unclear and Prnp-/- mice in which intergenic splicing with Prnd does not occur all develop normally with no obvious phenotypic abnormalities. Previous reports have presented phenotypes associated with the ablation of PrPc, e.g. PrP having copper dependent superoxide dismutase-activity (Brown et. al., 1997), attenuation of long-term potentiation in Prnp-/- mice (Collinge et al., 1994). However, some of these suggestions have been disputed (Waggoner et al. 2000; Hutter et al. 2003, Curtis et al., 2003). Additonally, other reported phenotypes, such as irregular circadian rhythms (Tobler et al. 1996) involve complex physiological processes and do not provide clarification of the physiological role of PrPc.
In order to learn more about the physiological function(s) of PrPc and potential pathways it might be involved in, we have utilised a murine neuroblastoma cell line (N2aPK1), which is highly susceptible towards prion infection and shows a high level of PrP expression, to create a stable cell line in which substantial knockdown of PrPc in an inducible and controlled manner can be achieved by shRNA technology. This approach allows comparison of expression differences in a homogenous cell population, in contrast to whole organs where differential expression may be "diluted out".
Using high density oligonucleotide microarray analysis of transcript expression profiles and also proteomics using the ICAT-technology (Gigy et al., 1999) we are utilising this cell line (termed PK1-TetR-shRNAPrnp) to identify genes and proteins that are differentially expressed upon controlled knockdown of PrPc by. Identification of transcripts and proteins appearing differentially expressed as a result of the knock-down of PrPc will provide valuable insights into the physiological function of PrP.
AD Jan Kranich, Gino Miele, Harald Seeger, Adriano Aguzzi, Universitätsspital Zürich, Switzerland
SP englisch
PO Deutschland