Poly(ADP-ribose) immunostaining to detect apoptosis induced by a neurotoxic fragment of prion protein.

摘要: PrP106-126 is a synthetic peptide representing codons 106-126 of the prion protein, which spontaneously forms amyloid fibrils and exerts neurotoxic effects on primary mouse brain cell cultures. Neurotoxicity by this commonly used as model for neurotoxicity observed in diseases involves formation reactive oxygen species which, turn, can cause DNA damage, including strand breaks. Strand breaks nuclear activate poly(ADP-ribose) polymerase to covalently modify proteins with poly(ADP-ribose). We, therefore, examined immunofluorescence whether or not triggers formation. We strong signals fraction cells, typically arranged clustered pattern, 30-48 h after addition. A few positive cells were also present untreated Cell morphology was suggestive apoptosis, confirmed positivity terminal deoxynucleotidyltransferase-mediated dUTP nick-end labelling (TUNEL) assay. On other hand, our assay did detect any 'early' activation morphologically normal that could have resulted from peptide-induced species. conclude immunostaining convenient reliable method visualizing undergoing apoptosis induced PrP106-126.