NR ATGW
AU Witusik,M.; Hulas-Bigoszewska,K.; Zawlik,I.; Golanska,E.; Piaskowski,S.; Szybka,M.; Liberski,P.P.; Rieske,P.
TI Neuronal differentiation of PC12 cells is defected by siRNA molecules inhibiting expression of Doppel protein
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-02
PT Konferenz-Poster
AB
Prnd is the gene encoding Doppel (Dpl) protein, which definite role has not been established yet. The present study was performed on nerve growth factor (NGF)-responsive line of PC12 rat pheochromocytoma cells, which undergo neuronal differentiation and develop into cells with morphological, electrophysiological, and neurochemical properties of sympathetic neurons. We used reverse transcription-coupled PCR method to determine the expression pattern of Prnd gene in relation to differentiation.
Treatment of PC12 cells with NGF resulted in significant increase of Dpl mRNA level, which may suggest a participation of this protein in neuronal differentiation process. In order to investigate this possibility, the expression of Dpl protein in PC12 cells was inhibited using specific siRNAs molecules active to Prnd as a target gene. We revealed that transfection of PC12 with above mentioned siRNAs abolished certain features of neuronal differentiation typically presented by PC12, during exposure to NGF. Immunocytochemical analysis demonstrated that silencing of Prnd gene resulted in down-regulation of neuronal markers: tyrosine hydroxylase and b III tubulin. These data indicate that typical pattern of PC12 neuronal differentiation is disturbed in absence of Dpl expression. For these observations, we concluded that Dpl protein may be important for the process of neuronal differentiation.
(The study was partially supported by KBN grant No. 3P05A11423)
AD M.Witusik, K.Hulas-Bigoszewska, I.Zawlik, E.Golanska, S.Piaskowski, M.Szybka, P.P.Liberski, P.Rieske, Department of Molecular Pathology and Neuropathology, Medical University of Lodz, Poland
SP englisch
PO Deutschland