DNA fragmentation follows delayed neuronal death in CA1 neurons exposed to transient global ischemia in the rat.
Apoptosis is an active, gene-directed process of cell death in which early fragmentation of nuclear DNA precedes morphological changes in the nucleus and, later, in the cytoplasm. In ischemia, biochemical studies have detected oligonucleosomes of apoptosis whereas sequential morphological studies show changes consistent with necrosis rather than apoptosis. To resolve this apparent discrepancy, we subjected rats to 10 minutes of transient forebrain ischemia followed by 1 to 14 days of reperfusion. Parameters evaluated in the CA1 region of the hippocampus included morphology, in situ end labeling (ISEL) of fragmented DNA, and expression of p53. Neurons were indistinguishable from controls at postischemic day 1 but displayed cytoplasmic basophilia or focal condensations at day 2; some neurons were slightly swollen and a few appeared normal. In situ end labeling was absent. At days 3 and 5, approximately 40 to 60% of CA1 neurons had shrunken eosinophilic cytoplasm and pyknotic nuclei, but only half of these were ISEL. By day 14, many of the necrotic neurons had been removed by phagocytes; those remaining retained mild ISEL. Neither p53 protein nor mRNA were identified in control or postischemic brain by in situ hybridization with riboprobes or by northern blot analysis. These results show that DNA fragmentation occurs after the development of delayed neuronal death in CA1 neurons subjected to 10 minutes of global ischemia. They suggest that mechanisms other than apoptosis may mediate the irreversible changes in the CA1 neurons in this model.
Petito
C K
CK
Department of Pathology, University of Miami School of Medicine, Miami, Florida 33136, USA.
Torres-Munoz
J
J
Roberts
B
B
Olarte
J P
JP
Nowak
T S
TS
Pulsinelli
W A
WA
D000818:Animals; D002545:Brain Ischemia / Q000473:pathology*; D016923:Cell Death; D004249:DNA Damage; D006624:Hippocampus / Q000473:pathology*; D008297:Male; D009474:Neurons / Q000473:pathology*; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors