The Journal of Neuroscience, March 15, 1999, 19(6):1988-1997
Ischemia model. All experimental procedures were approved by the Subcommittee on Animal Studies of the Veterans Affairs Medical Center (San Diego, CA). Male Wistar rats (250-300 gm) were fasted overnight. Anesthesia was induced with 3% halothane followed by maintenance with 1-2% halothane in an oxygen/nitrous oxide (30/70%) gas mixture. Catheters were inserted into the external jugular vein, tail artery, and tail vein to allow blood sampling, arterial blood pressure recording, and drug infusion. Both common carotid arteries were exposed and encircled by loose ligatures. Fifteen minutes before ischemia induction and 15min after ischemia, blood gases were measured and adjusted to PaO2 >90 mmHg and PaCO2 35-45 mmHg, pH 7.35-7.45, by adjusting tidal volume of the respirator. Bipolar EEG was recorded every 5-10 min before ischemia, continuously during the ischemic insult, and every 5min after ischemia until the rat recovered from the anesthesia. At the beginning of a 30min steady-state period before induction of ischemia, the inspired halothane concentration was decreased to 0.5%, and 150IU/kg heparin was administered intravenously. Blood was withdrawn via the jugular catheter to produce a mean arterial blood pressure of 50mmHg, and ischemia was induced by clamping both carotid arteries. Blood pressure was maintained at 50mmHg during the ischemic period by withdrawing or infusing blood through the jugular catheter. At the end of the ischemic period, the clamps were removed, and the blood was reinfused through the jugular catheter, followed by 0.5ml of 0.6M sodium bicarbonate. In all experiments, temperature was maintained at 37°C before, during, and after ischemia (15min of reperfusion). Halothane was discontinued at the end of ischemia, and all wounds were sutured. At 4or 24hr, 3d, or 1week after the ischemic episode, the animals were reanesthetized, tracheotomized, and artificially ventilated. For electron microscopic studies, the brains were perfused with ice-cold 2% paraformaldehyde and 2.5% glutaraldehyde in 0.1M cacodylate buffer. Sham-operated control rats were subjected to the same surgical procedures but without induction of ischemia.
The Journal of Neuroscience, January 1, 1998, 18(1):205-213
Figure 5. Histological analysis after 24hr of MCA occlusion. A, Photomicrograph showing the histological changes after 24hr of MCA occlusion in wild-type (Wt) and knock-out mutant mice (Sod2 /+). The infarct area was localized in the caudoputamen and MCA territory cortex in both mice groups. However, cortical infarction extended to the boundary zone of the anterior cerebral artery territory, and brain swelling was extremely severe in the knock-out mutant mice. Also shown are infarct volume (B) and hemisphere enlargement (C) in wild-type and knock-out mutant mice at 24hr ischemia. Values are mean±SE; **p<0.01and ***p<0.001,Student's t test. Cerebral infarction and hemisphere enlargement were significantly more severe in knock-out than in wild-type mice. Scale bar, 1 mm.
The Journal of Neuroscience, March 17, 2004, 24(11):2750-2759
Ischemic preconditioning and global ischemia. Age-matched male Sprague Dawley rats (Charles River Laboratories, Wilmington, MA) were maintained in a temperature- and light-controlled environment with a 14/10 hr light/dark cycle and were treated in accordance with the principles and procedures of the National Institutes of Health Guidelines for the Care and Use of Laboratory Animals. All protocols were approved by the Institutional Animal Care and Use Committee of the Albert Einstein College of Medicine. Animals were subjected to global ischemia by four-vessel occlusion (4 min for preconditioning; 10 min for global ischemia), followed by reperfusion as described (Kinouchi et al., 1993; Calderone et al., 2003). For sham operation, animals were subjected to the same anesthesia and surgical exposure procedures, except that the carotid arteries were not occluded. Body temperature was monitored and maintained close to 37.5 ± 0.5°C with a rectal thermistor and heat lamp until the animal had fully recovered from anesthesia. The rare animals that exhibited obvious behavioral manifestations (abnormal vocalization when handled, generalized convulsions, loss of >20% body weight by 3–7dor hypoactivity) and animals that failed to show complete loss of the righting reflex from 2 min after occlusion was initiated and anesthesia was discontinued until the end of occlusion were excluded. Histology and neuronal counts from six to eight rats per group (four sections per animal) were as described (Calderone et al., 2003). Statistical analysis was assessed by ANOVA, followed by Scheffe's post hoc tests. Ischemic preconditioning and global ischemia in gerbils were as described (Tanaka et al., 2002).
