H. Chigasaki

Repeated hyperbaric oxygen induces ischemic tolerance in gerbil hippocampus

Hyperbaric oxygen (HBO; 100% oxygen at 2 atmospheres absolute) was administered for 1 h to male Mongolian gerbils either for a single session or every other day for five sessions. Two days after HBO pretreatment, the gerbils were subjected to 5 min of forebrain ischemia by occlusion of both common carotid arteries under anesthesia. Seven days after recirculation, neuronal density per 1-mm length of the CA1 sector in the hippocampus was significantly better preserved in the five-session HBO pretreatment group (n = 10: 175.7 (47.8/mm, 54.9% of normal) than in the ischemic control group (n = 10: 26.2 (11.6/mm, 8.0% of normal) and in the single-session HBO pretreatment group (n = 7: 37.3 (21.7/mm, 11.4% of normal). Immunohistochemical staining for the 72-kDa heat-shock protein (HSP-72) in the CA1 sector performed 2 days following pretreatment revealed that the five-session HBO pretreatment increased the amount of HSP-72 present compared with that in the ischemic control group and in the single HBO pretreatment group. These results suggest that tolerance against ischemic neuronal damage was induced by repeated HBO pretreatment, which is thought to occur through the induction of HSP-72 synthesis.

Selective vulnerability of hippocampal CA3 neurons to hypoxia after mild concussion in the rat.

Immunohistochemical staining for microtubule-associated protein 2 (MAP2) and synaptophysin was used to investigate the effect of hypoxia on hippocampal neurons after mild concussion in the rat. Male Sprague-Dawley rats were divided into four groups: Group 1 (n = 3) was subjected to a mild impact-acceleration closed head injury, group 2 (n = 3) was subjected to 30 min of moderate hypoxia, group 3 (n = 5) was subjected to head trauma followed by 30 min of moderate hypoxia, and group 4 (n = 3) comprised sham-operated controls. All rats were fixed by transcardial perfusion 24 h after insult. No damage was observed in CA1 or CA2 neurons in any of the rats. However, rats in group 3 showed selective damage of hippocampal CA3 neurons manifested by a pycnosis and a marked decrease in MAP2 immunoreactivity. Presynaptic terminals visualized by synaptophysin immunostaining showed no differences among groups. The loss of immunoreactivity for the post-synaptic somal and dendritic protein marker MAP2 from the CA3 subfield 24 h after combined insults indicates an increased vulnerability of pyramidal cells in this brain area.

The effect of MK-801 on extracellular neuroactive amino acids in hippocampus after closed head injury followed by hypoxia in rats

Increased neuronal vulnerability to ischemia or hypoxia has been demonstrated following traumatic brain injury but not explained. Animal data suggest that neuronal damage after traumatic brain injury is caused mainly by massive glutamate release that activates N-methyl-D-aspartate (NMDA) receptors. Using rat models with controlled closed head injury (CHI) followed by hypoxia, we investigated extracellular concentrations of neuroactive amino acids in the hippocampus by in vivo microdialysis. CHI alone produced an immediate increase of glutamate and taurine; hypoxia alone did not alter amino acid concentrations. CHI followed by hypoxia produced a biphasic increase in extracellular glutamate and taurine, with an immediate peak after CHI and a prolonged plateau after hypoxia. Though changes in gamma-aminobutylic acid (GABA) concentration is also prolonged by combined traumatic and hypoxic insults, it showed less alteration than glutamate. Pre-treatment with dizocilpine maleate (MK-801), a non-competitive NMDA antagonist, did not affect the immediate peak of glutamate after CHI but significantly diminished the prolonged plateau after hypoxia. These findings suggest that traumatic brain injury may increase hypoxic release of glutamate, contributing to increased vulnerability to hypoxia. Our data suggest that MK-801 may be beneficial in preventing secondary neuronal damages by hypoxia.

Calcium Accumulation following Middle Cerebral Artery Occlusion in Stroke-Prone Spontaneously Hypertensive Rats

Delayed neuronal damage in the ischemic region of the rat brain following middle cerebral artery (MCA) occlusion in stroke-prone spontaneously hypertensive rats was studied. The distribution of neuronal damage was determined by 45Ca autoradiography. Accumulation of 45Ca was observed in the corpus callosum and ipsilateral cerebral cortex immediately following MCA occlusion. After 3 days of occlusion, 45Ca had accumulated in the ipsilateral pyramidal tract, the ventral posterior nucleus of the thalamus, and the lateral portion of the striatum. Significant accumulation of 45Ca was observed in the same areas after 7 and 14 days of occlusion. Next the effect of MK-801 on accumulation of 45Ca after MCA occlusion was examined using the same technique. MK-801 (0.5–10 mg/kg i.v.) or saline was administered 15 min before MCA occlusion, and volumes of accumulation of 45Ca were calculated 1 week after ischemic insults. MK-801 significantly reduced 45Ca uptake in the cortex, striatum, and thalamus. Furthermore, there was a strong statistical correlation between the volume of accumulation of 45Ca in the cortex and that in the thalamus (r = 0.8974; p < 0.001; n = 25). We speculate that delayed neuronal damage in the corpus callosum, ipsilateral pyramidal tract, and thalamus may be caused by secondary neuronal degeneration. However, neuronal damage in the striatum, a segment not supplied by the MCA, may be related to excessive release of glutamate.

Cerebral Blood Flow and Glucose Metabolism of the Ischemic Rim in Spontaneously Hypertensive Stroke-Prone Rats with Occlusion of the Middle Cerebral Artery

To determine acute postischemic metabolic changes of the ischemic rim under conditions of poor collateral circulation, we examined cerebral blood flow and glucose metabolism in the area of the brain around the ischemic tissue in 36 male spontaneously hypertensive stroke-prone rats (SHRSP) in the acute stage of focal ischemia. The right middle cerebral artery (MCA) was occluded dorsal to the rhinal fissure. Four hours after occlusion, local cerebral blood flow (LCBF), glucose content (LCGC), and glucose utilization (LCGU) were measured by quantitative autoradiographic techniques. The lumped constant was determined from the corresponding LCGC. LCBF showed a widespread and marked decrease in the cortex surrounding the ischemic core, in the thalamus, and in the medial portion of the striatum in the MCA-occluded hemisphere, while the lateral segment of the striatum showed an increase of 36%, compared with findings on the contralateral side. LCGC showed little regional variation, but there was an increase of 38% in the zone bordering the ischemic area. LCGU at the cortex surrounding the ischemic core and in the external capsule showed an increase of 55%. The cortex surrounding the ischemic core, the thalamus, and the lateral segment of the striatum in the MCA-occluded hemisphere showed significant decreases in LCGU. It has been speculated that a high accumulation of glucose reflects a demand for glucose for anaerobic glycolysis in the border areas and that such a demand is probably greater in cases of impaired oxygen delivery due to the presence of microcirculatory disturbances in the MCA-occluded SHRSP. The enhancement of glucose consumption may reflect anaerobic glycolysis. Because the hypermetabolic band was present in the cortex and the white matter, hypermetabolism of the white matter may be related to the glial cell.

Hepatocyte growth factor reduces infarct volume after transient focal cerebral ischemia in rats.

Hepatocyte growth factor (HGF) was originally discovered as a powerful mitogen for hepatocytes. HGF functions both as a neurotrophic factor as well as an angiogenetic factor. Furthermore, HGF has an anti-apoptotic effect on vascular endothelial cells. The present study examined the neuroprotective effect of HGF after transient focal cerebral ischemia in rats, in which an anti-apoptotic and an angiogenetic effect of HGF was assumed to contribute to the reduction of the infarct volume. The intraventricular administration of human recombinant HGF (90 micrograms) significantly reduced the infarct volume after 120 minutes occlusion of both the right middle cerebral artery (MCA) and the bilateral common carotid arteries (CCAs). In a separate series of experiments, we investigated both the anti-apoptotic effect on neurons and the angiogenetic effect of HGF histopathologically. The number of survival neurons and vascular lumina in the HGF group were significantly higher than those in the vehicle group. A large number of TUNEL positive neurons were observed in the inner boundary of the infarct area in the vehicle group, whereas only a few TUNEL positive neurons were observed in a corresponding area in the HGF group. In the HGF group, Bcl-2 protein was obviously represented in survival neurons as well as in vascular endothelial cells and in glial cells subjected to ischemia. These data suggest that HGF prevents apoptotic neuronal cell death by upregulating the production of Bcl-2 protein and by an angiogenetic effect in the central nervous system which affected transient focal cerebral ischemia.

Intraischemic hypothermia during pretreatment with sublethal ischemia reduces the induction of ischemic tolerance in the gerbil hippocampus.

We examined whether mild brain hypothermia during pretreatment with sublethal 2-min ischemia affected the tolerance to subsequent lethal 5-min ischemia. The neuronal densities in the hippocampal CA1 sector of gerbils preconditioned at mild brain hypothermia (32% of normal) were significantly lower than those in gerbils preconditioned at brain normothermia (70% of normal). 72-kDa heat-shock protein immunoreactivity in the CA1 sector preconditioned at mild hypothermia was reduced. These results suggest that mild brain hypothermia during pretreatment with sublethal ischemia reduces the tolerance to subsequent lethal ischemia.

Blood-Brain Barrier, Cerebral Blood Flow, and Cerebral Plasma Volume Immediately After Head Injury in the Rat

The purpose of the present study was to determine blood-brain barrier (BBB) permeability, regional cerebral blood flow (rCBF) and regional cerebral plasma volume (rCPV) in the period immediately after head injury, and thereby to evaluate the effects of vascular factors in the pathophysiology of traumatic brain injury. Male Sprague-Dawley rats (350-450 g) anesthetized with 1.0-1.5% halothane were subjected to an impact acceleration closed head injury at the moderate level. BBB permeability (n = 5), rCBF (n = 8) and rCPV (n = 9) were measured by quantitative autoradiographic techniques using 14C-alpha-aminoisobutyric acid (AIB), 14C-iodoantipyrine and 14C-sucrose, respectively. Intravenous administration of each radiotracer was simultaneous with the traumatic impact. At 10 min after injury, BBB permeability, the transfer constant for AIB, was less than 0.1 ml/kg/min for all regions except for those with a relatively leaky BBB. At 30 s after injury, a significant and heterogeneous increase in rCBF was observed at 9 subcortical regions (p < 0.05). RCPV increased significantly in the frontal cortex, parietal cortex, thalamus, and hypothalamus (p < 0.05). In our closed head injury model without severe hypertension, BBB disruption did not occur immediately after trauma. Vascular responses in the period immediately after trauma may result from the derangement of cerebral autoregulation.

In Vivo Studies of Extracellular Metabolites in the Striatum After Distal Middle Cerebral Artery Occlusion in Stroke-Prone Spontaneously Hypertensive Rats

BACKGROUND AND PURPOSE We demonstrated in a previous study that 45Ca accumulation in the lateral part of the striatum was detected 3 days after distal middle cerebral artery (MCA) occlusion using a 45Ca autoradiographic technique in stroke-prone spontaneously hypertensive rats. However, the mechanism of delayed neuronal damage that occurred in the lateral part of the striatum is unknown. We examined changes in amino acids and monoamines in the striatums of rat brains after MCA occlusion in stroke-prone spontaneously hypertensive rats using an in vivo brain microdialysis technique. METHODS Microdialysis probes were inserted into the lateral or medial part of the striatum 24 hours before the experiment. The dialysis probe was perfused continuously at 2 microL/min with Ringers solution, and the dialysate samples were collected every 20 minutes. After a 3-hour period for baseline stabilization, the right MCA was occluded. The dialysate count of monoamines and amino acids was determined by high-performance liquid chromatography. RESULTS After MCA occlusion, a threefold transient increase in glutamate was observed in the lateral part of the striatum. The level returned to its baseline value 60 minutes after MCA occlusion. Dopamine in the lateral part increased twofold to its peak value. This release persisted for 2 hours after MCA occlusion. There were no significant changes in these components in the extracellular fluid of the medial part of the striatum. CONCLUSIONS Our study demonstrated that changes of neurotransmitters in the lateral part of the striatum after MCA occlusion differed from those in the medial part. These results suggest that excessive release of glutamate and dopamine is related to delayed neuronal damage that occurs in the lateral part of the striatum in this model.

Resolution of peritumoral brain edema following excision of meningioma.

We studied the resolution of peritumoral brain edema after meningioma excision. In twenty-nine patients with meningioma, the total volume of tumor and the peritumoral edema were measured planimetrically by serial CT scans and MRI with or without contrast enhancement. Four different patterns of postoperative resolution of hypodense volume on CT were observed: Group A: a large hypodensity rapidly decreased and disappeared, which may be related to the clearance of the real peritumoral edema in meningioma. Group B: a small hypodensity gradually disappeared. Group C: the hypodensity remained unchanged, which may result from the damaged brain tissue. Group D: the hypodensity progressively decreased but persisted, which may represent both the peritumoral edema and damaged brain tissue. We have calculated the resolution rate of edema fluid using the clearance curve of Groups A and D. The average resolution rate of edema fluid during the passage through 1 cm3 of the peritumoral white matter was 0.0493 ml/day. We speculate that 50% of edematous white matter, which presented as hypodensity on a CT scan, may be resolved in 4 days after total removal, and that 90% may be resolved in 14 days.