Mechanisms of delayed cell death following hypoxic-ischemic injury in the immature rat: evidence for apoptosis during selective neuronal loss.
The mechanisms leading to delayed cell death following hypoxic-ischemic injury in the developing brain are unclear. We examined the possible roles of apoptosis and microglial activation in the 21-day-old rat brain following either mild (15 min) or severe (60 min) unilateral hypoxic-ischemic injury. The temporal and spatial patterns of DNA degradation were assessed using gel-electrophoresis and in-situ DNA end-labelling. Microglial activation, mitochondrial failure and cell death were examined using lectin histochemistry, 2,3,5,triphenyl-H-tetrazolium chloride (TTC) staining and acid fuchsin staining, respectively. Selective neuronal death produced by the 15 min injury was associated with the development of apoptotic morphology, DNA laddering and acidophilia from 3 days post-hypoxia. The 60 min injury accelerated this process with some cells showing signs of DNA degradation at 10 h post-hypoxia. However, in the cortex, which developed infarction after the 60 min injury, a different pattern of cell loss occurred. The DNA and mitochondria remained intact, and cells basophilic, until after 10 h post-hypoxia, then widespread necrosis developed by 24 hr. In contrast to regions of selective neuronal loss, DNA degradation was initially random (at 24 hr), with 180bp DNA ladders not detected until 3 days post-hypoxia. There was no morphological evidence of apoptosis. Microglial activation coincided with the onset of DNA degradation in regions of selective neuronal loss but not infarction, suggesting a possible role in selective neuronal death. The results suggest that cortical infarction, which was delayed for at least 10 h, was necrotic, and occurred independently of microglial activation and apoptosis. In contrast, selective neuronal death was apoptotic.
Beilharz
E J
EJ
Research Centre for Developmental Medicine and Biology, Auckland, New Zealand.
Williams
C E
CE
Dragunow
M
M
Sirimanne
E S
ES
Gluckman
P D
PD
D000818:Animals; D000831:Animals, Newborn / Q000502:physiology*; D017209:Apoptosis; D002545:Brain Ischemia / Q000473:pathology* / Q000503:physiopathology; D016923:Cell Death; D003851:Deoxyribonucleases; D000860:Hypoxia / Q000473:pathology* / Q000503:physiopathology; D017628:Microglia / Q000473:pathology; D008928:Mitochondria / Q000502:physiology; D009474:Neurons / Q000473:pathology*; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors
D003851:Deoxyribonucleases