Impairment of mitochondrial respiration after cerebral hypoxia-ischemia in immature rats: relationship to activation of caspase-3 and neuronal injury.
Mitochondrial damage may play a key role in the development of necrotic and apoptotic hypoxic-ischemic (HI) brain damage. It has previously been shown that mitochondrial respiration is depressed in the cerebral cortex after HI in neonatal animals. The aim of the present study was to further characterize the time course of the mitochondrial impairment during reperfusion and the correlation between the respiratory control ratio and brain injury and activation of caspase-3. Rat pups were subjected to unilateral carotid artery ligation and exposed to hypoxia (7.7% oxygen). Mitochondrial respiration was measured 0-72 h after HI in a mitochondrial fraction isolated from cerebral cortex. Microtubule associated protein-2 (MAP2) and caspase-3 were analyzed with immunoblotting in cerebral cortex homogenates. In addition, the time course of caspase-3 activation was measured as DEVD cleavage. The mitochondrial respiratory control ratio in cerebral cortex decreased immediately after HI followed by a partial recovery at 3-8 h. Thereafter, a secondary drop occurred with a minimum reached at 24 h of reperfusion. The secondary loss of respiratory function was accompanied by depletion of MAP2, cleavage of caspase-3 and an increased caspase-3 -like activity at 3-24 h after the insult. In conclusion, the primary phase of mitochondrial dysfunction was paralleled by a moderate decrease of MAP2 and a limited activation of caspase-3. The secondary mitochondrial impairment was associated with neuronal injury and pronounced activation of caspase-3.
Puka-Sundvall
M
M
Department of Anatomy and Cell Biology, Perinatal Center, Göteborg University, S-405 30, Göteborg, Sweden.
Wallin
C
C
Gilland
E
E
Hallin
U
U
Wang
X
X
Sandberg
M
M
Karlsson
J
J
Blomgren
K
K
Hagberg
H
H
D000818:Animals; D000831:Animals, Newborn; D000918:Antibody Specificity; D015153:Blotting, Western; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism*; D019069:Cell Respiration / Q000502:physiology; D002540:Cerebral Cortex / Q000473:pathology; D004789:Enzyme Activation / Q000502:physiology; D005260:Female; D020925:Hypoxia-Ischemia, Brain / Q000378:metabolism* / Q000473:pathology; D008297:Male; D008869:Microtubule-Associated Proteins / Q000032:analysis / Q000276:immunology; D008928:Mitochondria / Q000378:metabolism*; D009474:Neurons / Q000737:chemistry / Q000201:enzymology* / Q000473:pathology; D051381:Rats; D011920:Rats, Inbred WF
D008869:Microtubule-Associated Proteins; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases