Detection of early tissue injury in vivo using fluorescent cell death markers in rat dentate gyrus.
Fluorescent markers of cell death offer superior selectivity and sensitivity, although their applicability in detecting tissue injury under in vivo conditions remains uncertain. Here we examined whether ethidium bromide and Hoechst 33342, two widely used markers for cell necrosis and apoptosis, can be used in vivo to detect different types of cell death induced by Na,K-ATPase inhibition. Microinfusion of fluorescent markers and ouabain was made unilaterally into adult rat dentate gyrus. It was found that, at different time points post-injury, dentate cells that were exposed to ouabain but not to vehicle control showed marked loss of membrane integrity and exhibited nuclear condensation, as revealed by ethidium bromide and Hoechst 33342 staining, respectively. However, this pattern of cell death was not associated with DNA fragmentation and formation of apoptotic bodies, suggesting involvement of atypical cell apoptosis.
Omar
A I
AI
Neuroscience, Ottawa Health Research Institute, Ottawa Hospital and University of Ottawa, Ontario, K1Y 4K9, Ottawa, Canada.
Hu
B
B
D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D015415:Biomarkers / Q000032:analysis; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D018891:Dentate Gyrus / Q000737:chemistry* / Q000187:drug effects / Q000473:pathology*; D005456:Fluorescent Dyes / Q000032:analysis*; D008297:Male; D010042:Ouabain / Q000494:pharmacology; D051381:Rats; D020318:Rats, Long-Evans; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors
D015415:Biomarkers; D005456:Fluorescent Dyes; D010042:Ouabain; D000254:Sodium-Potassium-Exchanging ATPase