A modified four-vessel occlusion model for inducing incomplete forebrain ischemia in rats.

Abstract

The four-vessel occlusion (4VO) model of Pulsinelli and Brierley (Stroke 1979; 10:267-272) has been modified for use in halothane-nitrous oxide-anesthetized, physiologically controlled rats that were ventilating spontaneously. Selection criteria for the classification of severity of ischemia were established by correlating changes in the electroencephalogram and the general physiological status with measurements of regional blood flow and regional energy metabolism. In 13% of animals, 4VO did not cause flattening of the electroencephalogram, and such animals were classified as undergoing only "oligemia." In 65% of rats, the electroencephalogram flattened and blood pressure sharply increased with 4VO, whereas spontaneous respiration continued. This group exhibited almost complete ischemia in autoradiographic blood-flow studies, severe acidosis, and depletion of adenosine 5'-triphosphate and glucose in the forebrain and, hence, was classified as the "ischemia" group. The remaining 22% stopped breathing after vascular occlusion and were rejected for further study. Survival experiments of ischemic animals revealed the typical postischemic sequelae, with primary metabolic recovery after 8 hours of recirculation in all brain structures followed after 8-24 hours by severe biochemical deterioration and neuronal death in the striatum and hippocampus. Postischemic seizure activity was rare. The main advantages of the present modification in comparison with the original method are 1) the application of anesthesia without loss of primary selection criteria, 2) the possibility of invasive physiological monitoring, and 3) the absence of postischemic seizures, which eliminates the necessity for secondary selection criteria. (Stroke 1989;20:938-946) I n 1979, Pulsinelli and Brierley 1 described a model of incomplete forebrain ischemia in unanesthetized rats that was produced by occlusion of four major arteries supplying the brain. Four-vessel occlusion (4VO) was performed in two stages: 1) by coagulation of both vertebral arteries through the alar foramen and 2) after a delay of 1 day, by transient occlusion of both common carotid arteries.1 The severity of ischemia is judged by neurological investigation, and animals are then selected for further study on this basis. Received September 19, 1988; accepted February 2, 1989. ischemia by examining neurological signs. For this reason, we have established selection criteria of severe forebrain ischemia that can be applied to anesthetized, spontaneously breathing animals and that are compatible with postischemic survival.Materials and Methods We used 163 adult male Wistar rats of the HanWist strain (Zentralinstitut fur Versuchstiere, Hannover, FRG) (mean body weight, 310 g). On the first day of the experiment, rats were anesthetized in a jar flushed with 4% halothane in 70% nitrous oxide-30% oxygen until unresponsive, placed on a rat operating table (Hugo Sachs, Freiburg, FRG), and maintained under anesthesia with a face mask surrounded by a larger, semiclosed container connec...

Glial Expression of the β-Amyloid Precursor Protein (APP) in Global Ischemia

“…A 30-min global ischemia was achieved with the four vessel occlusion described by Pulsinelli and Brierley (1979), as modified by Schmidt-Kastner et al (1989). Adult male Wistar rats weighing 200-350 g were anesthe tized with halothane/nitrous oxide.…”

“…The present study investigates immunocytochemi cally the expression of glial APP in different ana tomical regions and at different time points after global cerebral ischemia. The four-vessel occlu s ion model used here (Pulsinelli and Brierley, 1979;Schmidt-Kastner et a!. , 1989 leads to neuro nal degeneration in the dorsolateral striatum within 12 h, while the neuronal degeneration in the hippo campal CAl region occurs with a characteristic de lay of 2-3 days.…”

rAAV8-733-Mediated Gene Transfer of CHIP/Stub-1 Prevents Hippocampal Neuronal Death in Experimental Brain Ischemia

“…We used the four-vessel occlusion model to induce transient global brain ischemia in rats as previously described, 37,38 with slight modifications. Briefly, both rAAV-infected male rats (injected at postnatal day 60) and non-infected controls at postnatal day 90 were anesthetized with a mixture of oxygen and isoflurane (model 72-6474, Harvard Systems) and had anesthesia sustained by a mask attached to the system.…”

Prevention of in Vivo Excitotoxicity by a Family of Trialkylglycines, a Novel Class of Neuroprotectants

“…For the transient forebrain ischemia experiments, we used the four-vessel occlusion model of Pulsinelli and Brierley (1979) with the modifications of Schmidt-Kastner et al (1989). Briefly, rats were anesthetized with 120 g/g ketamine and 15 g/g xylazine and placed in a rat holder.…”