Focal cerebral infarction in the rat: II. Neuropathological study and local cerebral blood flow pattern

We have recently reported that middle cerebral artery (MCA) occlusion in the rat produces a uniform pattern of cerebral ischemia in an acute phase. This study was done to determine if this model is also useful for quantitative evaluation of infarction size in a chronic phase. [Methods] Sprague-Dawley rats were anesthetized with halothane and left MCA was occluded via transretro-orbital approach. The following studies were done. Neuropathological study was done one week after MCA occlusion. After perfusion fixation, the brain was cut into 6 coronal slices and stained sections were examined. Local cerebral blood flow patterns were observed by 14C-iodoantipyrine autoradiographic technique 1, 2, and 5 days after the occlusion. [Results] Neuropathological studies invariably showed infarct in the cortex and the lateral part of the basal ganlia. The ratio of the infarct to the total areas of both hemispheres in 6 coronal sections was 14.05 +/- 2.66% (Mean +/- SD) in MCA occluded animals (N = 14) and 0.59 +/- 0.46% in sham operated animals (N = 12). Relative to the contralateral hemisphere, marked reduction in CBF was seen in the territory of the MCA and moderate reductions were also seen in the surrounding areas. The same pattern of increased CBF as previously reported was also seen in the ipsilateral substantia nigra and globus pallidus 1, 2, and 5 days after the occlusion. These results indicate the usefulness of this chronic focal cerebral infarction model in the evaluation of infarction.     

Effect of the kappa-1 opioid agonist CI-977 on ischemic brain damage and cerebral blood flow after middle cerebral artery occlusion in the rat

The effects of the kappa-1 opioid agonist CI-977 upon the volume of ischemic brain damage (defined using quantitative neuropathology) and local cerebral blood flow (CBF) (defined using quantitative [¹⁴C]iodoantipyrine autoradiography) have been examined at 4 h and 30 min, respectively, after permanent middle cerebral artery (MCA) occlusion in halothane-anesthetised rats. Treatment with CI-977 (0.3 mg/kg, s.c.) 30 min before and 30 min after occlusion of the MCA reduced the volume of infarction in the cerebral hemisphere (reduced by 27% when compared to vehicle;P<0.05) and cerebral cortex (reduced by 32%;P<0.05), despite a marked and sustained hypotension, with only minimal effect on damage in the caudate nucleus. In the hemisphere contralateral to the occluded MCA, treatment with CI-977 (0.3 mg/kg, s.c.) 30 min prior to the induction of ischemia failed to demonstrate any significant effect on either the level of local CBF in any of the 25 regions examined or on the volume of low CBF determined by frequency distribution analysis. In the hemisphere ipsilateral to MCA occlusion, CI-977 failed to produce statistically significant alterations in either the level of local CBF in 23 of the 25 regions or on the volume of low CBF, but areas of hyperemia were observed in both the medial caudate nucleus and lateral thalamus (local CBF increased by 65% and 86%, respectively, when compared to vehicle). The results of the present study indicate that the kappa-1 opioid agonist CI-977 is neuroprotective in a rat model of focal cerebral ischemia where key physiological variables have been assessed throughout the entire post-ischemic period, and fail to demonstrate that the neuroprotective effects of CI-977 in this model are due to improved blood flow to ischemic tissue.     

Nimodipine and the haemodynamic and histopathological consequences of middle cerebral artery occlusion in the rat

The effect of the administration of nimodipine (1 microgram kg-1 min-1), initiated 5 min after occlusion of a middle cerebral artery (MCA), upon cerebral haemodynamics ([14C]iodoantipyrine autoradiography) and neuropathological outcome (volume of histologically ischaemic tissue) was investigated in anaesthetized rats. Measurements were made of the level of local CBF (LCBF) in a total of 37 neuroanatomically defined areas, either ipsilateral or contralateral to the occluded vessel, and the autoradiograms were examined using a new approach to quantitative densitometry that employed a frequency distribution analysis of the CBF in sections of the brain at different coronal planes. Both methods of analysis showed that nimodipine, administered after the ischemic event, did not modify the pattern of CBF distribution after MCA occlusion. The extent of ischaemic brain damage was determined by histological examination. There was no evidence that the extent of ischaemic damage, either in sections at eight different coronal planes or in overall volume, was significantly different in animals that received nimodipine from that observed in animals that received only the vehicle used to dissolve the drug. The lack of cerebral circulatory and neuropathological alterations when nimodipine administration is initiated after occlusion of the MCA is contrasted with the higher levels of LCBF and the reductions in the volume of ischaemic tissue that were found when nimodipine was administered before occlusion of the artery.     

Cortical infarct volume is dependent on the ischemic reduction of perifocal cerebral blood flow in a three-vessel intraluminal MCA occlusion/reperfusion model in the rat

Occlusion of the middle cerebral artery (MCA) causes a reduction of cerebral blood flow (CBF), which shows a progressive decrease from the periphery to the core of the MCA territory. The severity of ischemia is dependent on the duration of the ischemic episode and degree of CBF reduction. Fixing the ischemic episode to 1 h, we have examined whether or not cortical infarct size was related to the degree of CBF reduction in a perifocal cortical area in rats. One-hour intraluminal MCA occlusion accompanied with bilateral common carotid artery (CCA) occlusion (three-vessel occlusion/reperfusion model) was carried out in Sprague-Dawley rats and CBF was monitored with laser-Doppler flowmetry in the fronto-parietal cortex, an area which is perifocal to the core of the MCA territory. Finally, infarct size was measured 7 days later and was related to the corresponding CBF decrease. Sequential ipsilateral CCA, MCA and contralateral CCA occlusions produced reductions of CBF to 96%, 52% and 33% of baseline, respectively. Cortical infarct volume was found to be dependent on the corresponding reduction of perifocal cortical CBF during the ischemic episode. These results show that the reduction of CBF in the periphery of the MCA territory during 1-h focal ischemia determines infarct size in a three-vessel occlusion/reperfusion model.     

Quantitative EEG alterations after isovolemic-hemodilutional augmentation of cerebral perfusion in stroke patients

Eleven patients with ischemic neurologic deficits in the middle cerebral arterial (MCA) territory and ipsilateral slowing on initial EEG underwent venesection and equal volume replacement with intravenous 5% human serum albumin. As the mean hematocrit was reduced by 19%, the mean cerebral blood flow (CBF) in the MCA territory of the affected and contralateral hemisphere determined by the 133Xenon inhalation technique increased 18 and 21%, respectively. Similarly, CBF in the contralateral occipital region increased 17%. The percentage total slow-wave EEG activity (fractional sum of theta and delta activity, 1.0 to 7.5 Hz) determined by fourier analysis was reduced significantly in the affected MCA territory and in the contralateral occipital region within 1 to 2 hours after isovolemic hemodilution. Using quantitative EEG analysis, rapid improvement in background EEG activity can be demonstrated following the diffuse elevation in CBF by hemodilution.     

Monitoring the temporal and spatial activation pattern of astrocytes in focal cerebral ischemia using in situ hybridization to GFAP mRNA: comparison withsgp-2 andhsp70 mRNA and the effect of glutamate receptor antagonists

We investigated the temporo-spatial expression of astrocyte glial fibrillary acidic protein (gfap) and sulfated glycoprotein 2 (sgp-2) mRNAs in comparison to 70-kDa heat shock protein (hsp70) mRNA by in situ hybridisation in rats subjected to permanent occlusion of the middle cerebral artery (MCA).Gfap mRNA started to increase in the cingulate cortex of the lesioned hemisphere 6 h after MCA occlusion and gradually spread over the lateral part of the ipsilateral cortex and the striatum from 12 h to 3 days, peaking at 3 days after MCA occlusion.Gfap mRNA also increased in the contralateral cingulate cortex and corpus callosum at 12 and 24 h.Hsp70 mRNA increased markedly in the ipsilateral cortex adjacent to the ischemic lesion, and slightly within the lesion area from 3 to 24 h and disappeared after 3 days. By 7 days,gfap andsgp-2 mRNAs were increased markedly in the peri-infarct area, and in the ipsilateral thalamus parallel with the delayed neuronal damage, whereas the widespread increase ofgfap mRNA in the ipsilateral hemisphere declined. Post-occlusion treatment with the glutamate receptor antagonists MK-801 and NBQX slightly attenuated the induction ofgfap but did not qualitatively affect the topical expression pattern. Within the cingulate cortex MK-801 treatment resulted in a significant decrease of the signal intensity at all survival times, reflecting most likely an attenuation of lesion-induced spreading depression like depolarization waves by MK-801. The area ofhsp70 expression was reduced by both MK-801 and NBQX, most likely reflecting the decrease of the lesion area by both treatment regimens. Our study thus revealed an early and widespread increase ofgfap mRNA in the non-ischemic areas including the contralateral hemisphere starting between 3 and 6 h, and a delayed circumscribed expression in the peri-infarct border zone after 1 week. Comparison with the expression ofhsp70 mRNA suggests that the absence of an earlygfap mRNA induction in the peri-lesion zone reflects an impairment of astrocytic function which may be of importance for infarct growth during the early evolution of the pathological process.     

Acute improvement in histological outcome by MK-801 following focal cerebral ischemia and reperfusion in the cat independent of blood flow changes.

The present study reports on the acute effects of MK-801 on the histopathological outcome and blood flow changes during focal cerebral ischemia and reperfusion. In addition, acute changes in the EEG and blood pressure are also reported. In 16 halothane-anesthetized cats, the left middle cerebral artery (MCA) was occluded for 2 h followed by 4 h of reperfusion. Thirty minutes after the onset of ischemia, eight animals were treated with 1 mg/kg of MK-801, while eight animals received saline. Blood flow from the peripheral MCA territory was measured with H2 clearance. There was a comparable reduction in blood flow (down to 20% of control) in the ischemic gyri of the two groups followed by a partial recovery after recirculation. There was a similar decrease in the EEG amplitude over the ischemic central MCA territory in the treated and the untreated group. Treatment with MK-801 induced a burst suppression in the EEG and a transient drop (11.4 +/- 6.5 mm Hg) in the mean arterial pressure. The volume of early ischemic damage decreased by one-third in the MK-801-treated group compared to the untreated one, both in the total hemisphere (from 29 +/- 10 to 20 +/- 5%) and in the hemispheric cortex (range 36 +/- 8 to 24 +/- 13%). A major fraction of this improvement was localized to the middle and posterior parietal (mainly perifocal) regions of the MCA territory. These results show that in our model, MK-801 improves histopathological outcome despite the lack of apparent effect on the cortical blood flow, and an adverse effect on the systemic blood pressure. This is the first report that describes data on a reproducible model of reperfusion after temporary occlusion of the MCA in a cat, extending the findings of the Glasgow group, who observed similar neuroprotection in models of permanent MCA occlusion.     

Blood flow through cerebral collateral vessels one month after middle cerebral artery occlusion

Acute occlusion of a middle cerebral artery (MCA) reduces cerebral blood flow in normotensive Wistar-Kyoto rats (WKY) and in stroke-prone spontaneously hypertensive rats (SHRSP). The goal of this study was to determine whether MCA occlusion produces a sustained reduction in cerebral blood flow or whether collateral vessels restore blood flow to normal levels. We measured blood flow through cerebral collateral vessels to the territory of the occluded MCA and to homologous tissue of the other hemisphere in WKY 1 month after occlusion of the MCA. Cerebral blood flow, measured with microspheres, was restored to normal levels under control conditions in the territory of the occluded MCA. During vasodilatation produced by seizures, blood flow and vascular conductance were increased to similar levels in tissue distal to the site of MCA occlusion and in the homologous tissue of the other hemisphere. MCA occlusion did not produce infarction in any of the WKY. In contrast, 1 month after MCA occlusion in SHRSP, a large atrophic infarct was invariably present in the territory of the occluded MCA. The number of collateral vessels to the territory of the MCA do not differ in SHRSP and WKY. Internal diameter and orientation of the anastomotic vessels differ in SHRSP and WKY. We conclude that, after 1 month of MCA occlusion, changes in the collateral vessels supplying the territory of the occluded MCA in WKY were sufficient to restore blood flow to normal under control conditions and to virtually normal levels during vasodilatation.(ABSTRACT TRUNCATED AT 250 WORDS)     

Failure of the somatosensory evoked potential following middle cerebral artery occlusion and high-grade ischemia in the cat--effects of hemodilution

Acute focal ischemia was created in 10 cats by unilateral retro-orbital middle cerebral artery (MCA) occlusion. Regional cerebral blood flow (CBF) was determined utilizing the hydrogen clearance technique from electrode recordings within the gray matter and white matter of the ectosylvian gyrus of both hemispheres. The somatosensory evoked potential (SSEP) was obtained during contralateral median nerve stimulation. When the MCA was clipped the white and gray matter blood flows in the ipsilateral ectosylvian gyrus were reduced to 14.8 +/- 19.6% and 19.3 +/- 23.7% of control, and the cortical component of the SSEP was abolished. In the contralateral hemisphere an average increase of 3.5% above the control latency and a 10% mean depression in the amplitude of the cortical component of the SSEP were observed following occlusion. CBF in the contralateral hemisphere was unaffected by the MCA clip. Infusion of saline or dextran to lower the hematocrit by approximately 45% did not significantly improve blood flow or restore the SSEP in the hemisphere ipsilateral to the MCA clip. However, significant increases in the contralateral hemisphere gray matter CBF occurred following hemodilution while the latency of the cortical component of the SSEP in this same hemisphere was significantly extended. Elevations in gray and white matter blood flows were achieved in the experimental hemisphere of 3 of 10 cats suggesting a wide range of variation in the collateral circulation.     

Focal cerebral ischemia in rats: effect of hemodilution with alpha-alpha cross-linked hemoglobin on CBF.

Hemodilution has had limited success as a treatment of cerebral ischemia. When using a nonoxygen binding fluid, the therapeutic efficacy of hemodilution-induced increases in CBF are offset by concomitant decreases in oxygen content. The effect of hemodilution, with diaspirin alpha-alpha cross-linked hemoglobin (DCLHb), on CBF during middle cerebral artery occlusion was assessed. Rats were hemodiluted to one of the following hematocrits (Hct): (a) 44/Hct, (b) 37/Hct, (c) 30/Hct, (d) 23/Hct, (e) 16/Hct, or (f) 9/Hct. After 10 min of ischemia, CBF was determined with 14C-iodoantipyrine. Coronal brain sections were evaluated for areas with a CBF of 0-10 and 11-20 ml 100 g-1 min-1. In addition, oxygen delivery was calculated. In the center of the ischemic zone, both areas of low CBF were less in the 30/Hct, 23/Hct, and 16/Hct groups compared with the 44/Hct and 37/Hct groups; and both areas were less in the 9/Hct group compared with the other five groups (p < 0.05). For the hemisphere contralateral to occlusion, there was a direct correlation between hematocrit and oxygen delivery. However, for the hemisphere ipsilateral to occlusion, oxygen delivery increased as hematocrit decreased (44/Hct, 8.6 +/- 0.3 vs. 9/Hct, 13.6 +/- 0.4 [mean +/- SD, ml 100 g-1 min-1]). The results of this study support a hypothesis that hemodilution with DCLHb decreases the extent of focal cerebral ischemia.     

Protective effect of the glutamate antagonist, MK-801 in focal cerebral ischemia in the cat

The effects of the glutamate N-methyl-D aspartate (NMDA) receptor antagonist, MK-801, upon ischemic brain damage has been examined in anesthetized cats. Focal cerebral ischemia was produced by permanent occlusion of one middle cerebral artery and the animal were killed 6 h later. The amount of early ischemic damage was assessed in coronal sections at 16 predetermined stereotactic planes. Pretreatment with MK-801 (5 mg/kg, i.v.), 30 min before occlusion of the middle cerebral artery significantly reduced the volume of ischemic damage (from 32.7 +/- 4.0% of the cerebral hemisphere in vehicle-treated cats to 16.2 +/- 4.5% in MK-801-treated cats). NMDA receptor antagonists that penetrate the blood-brain barrier, such as MK-801, merit further study as protective agents against ischemic brain damage.     

Local cerebral glucose utilization in chronic middle cerebral artery occlusion in the cat

This study examines the correlation between local CMRglc (LCMRglc) alterations and clinicopathological changes in a chronic middle cerebral artery (MCA) occlusion model in the cat. The left MCA was occluded for a period of 2 h. The animals were grouped into mild, moderate, and severe ischemia based on the depression of the EEG 30 min after the MCA occlusion. Following release of the clip, the animals were allowed to recover for a week during which time daily neurological examinations were performed. On the seventh day [14C]2-deoxyglucose was injected for the determination of LCMRglc. Alternative blocks were processed for histological evaluation in which both neuronal and phagocytic changes were graded into four categories (0 = normal to 3 = severe). LCMRglc (mumol/100 g/min) in the ischemic hemisphere (all histological grades) was significantly lower than the metabolic rate in comparable regions of the sham MCA occlusion group. Regions with significant phagocytosis (grade 2 and 3) invariably exhibited activated glucose metabolism (57.4 +/- 8.4 and 105.9 +/- 6.8 mumol/100 g/min, respectively), which was significantly higher than in regions without phagocytosis (30.4 +/- 0.8 mumol/100 g/min). There was a significant gradient of metabolism in the central, peripheral, and boundary zone and the non-MCA territory in the animals with severe ischemic lesions. LCMRglc in the central MCA territory was well correlated with the EEG amplitude changes (r = 0.82, p less than 0.05) and the morphological score (r = -0.89, p less than 0.05). The metabolic rate was significantly depressed in both the ipsilateral and the contralateral central MCA territories in comparison with the sham occlusion animals. The depression in LCMRglc in the contralateral hemisphere correlated well with the concomitant depression in the contralateral EEG amplitude. These studies demonstrate that local heterogeneous metabolic alterations and contralateral cortical diaschisis exist chronically following temporary MCA occlusion and that the increases in local cerebral glucose metabolism seen in chronic stroke may be due to phagocytotic activity.     

Effect of pretreatment with the calcium antagonist nimodipine on local cerebral blood flow and histopathology after middle cerebral artery occlusion

We used the [¹⁴C]iodoantipyrine autoradiography technique to study the effect of pretreatment with the calcium antagonist nimodipine on local cerebral blood flow (1CBF) in rats that underwent middle cerebral artery (MCA) occlusion. In untreated control animals there were profound localized reductions in 1CBF 30 minutes after MCA occlusion. These were most pronounced in neocortical areas and in the caudate nucleus ipsilateral to the MCA occlusion. In animals pretreated with nimodipine (1 μg · kg^?1 · min^?1 for 30 minutes before and 30 minutes after MCA occlusion), the ipsilateral decrease in 1CBF in cortical regions was significantly less than that in control animals. The drug did not appear to alter 1CBF in the ipsilateral caudate nucleus. Neuropathological quantification of the ischemic damage present 3 hours after occlusion showed thai: nimodipine pretreatment reduced the volume and extent of cellular damage in the periphery but not in the core of the lesion.     

Local hemodynamic changes during transient middle cerebral artery occlusion and recirculation in the rat: a [C]iodoantipyrine autoradiographic study

We evaluated acute alterations of local cerebral perfusion following 30 min of transient right proximal middle cerebral artery (MCA) clip-occlusion in the rat and following two intervals of postischemic reperfusion. Local cerebral blood flow (1CBF) was assessed by [¹⁴C]iodoantipyrine autoradiography. Brain temperature was controlled at 35.5–36.5°C throughout the experiment. We measured ICBF in four groups of rats: (a) sham-operated controls (n = 5), (b), following 30 min MCA occlusion (n = 5), (c) following 30 min of MCA occlusion with 15-min reperfusion (n = 6) and (d) following 30 min of MCA with 120-min reperfusion (n = 6). 1CBF was measured in seven regions of the ischemic and non-ischemic hemispheres. MCA occlusion induced an ipsilateral reduction of 1CBF, which was most severe in the parietal cortex (8.4 ± 4.0% of control, mean ± S.D.), and dorsolateral caudoputamen (20.0 ± 13.4% of control). 1CBF in the non-ischemic hemisphere and in ipsilateral regions lying outside the MCA territory also decreased significantly. 1CBF recovery was incomplete when assessed following only 15 min of reperfusion. Reperfusion of 120 min led to return of cortical CBF to control levels, but 1CBF in the caudoputamen remained depressed (50–55% of control values). Caudoputaminal CBF and cortical CBF values were highly correlated with one another under normal and ischemic conditions, but this correlation was disrupted following reperfusion. On the basis of these results, we speculate that, if a means were found to enhance the early recovery of 1CBF following transient ischemia, this might expand the therapeutic window of opportunity for the institution of other neuroprotective strategies.     

Reversible middle cerebral artery occlusion without craniectomy in rats

To develop a simple, relatively noninvasive small-animal model of reversible regional cerebral ischemia, we tested various methods of inducing infarction in the territory of the right middle cerebral artery (MCA) by extracranial vascular occlusion in rats. In preliminary studies, 60 rats were anesthetized with ketamine and different combinations of vessels were occluded; blood pressure and arterial blood gases were monitored. Neurologic deficit, mortality rate, gross pathology, and in some instances, electroencephalogram and histochemical staining results were evaluated in all surviving rats. The principal procedure consisted of introducing a 4-0 nylon intraluminal suture into the cervical internal carotid artery (ICA) and advancing it intracranially to block blood flow into the MCA; collateral blood flow was reduced by interrupting all branches of the external carotid artery (ECA) and all extracranial branches of the ICA. In some groups of rats, bilateral vertebral or contralateral carotid artery occlusion was also performed. India ink perfusion studies in 20 rats documented blockage of MCA blood flow in 14 rats subjected to permanent occlusion and the restoration of blood flow to the MCA territory in six rats after withdrawal of the suture from the ICA. The best method of MCA occlusion was then selected for further confirmatory studies, including histologic examination, in five additional groups of rats anesthetized with halothane. Seven of eight rats that underwent permanent occlusion of the MCA had resolving moderately severe neurologic deficits (Grade 2 of 4) and unilateral infarcts averaging 37.6 +/- 5.5% of the coronal sectional area at 72 hours after the onset of occlusion.(ABSTRACT TRUNCATED AT 250 WORDS)     

The glutamate antagonist MK-801 reduces focal ischemic brain damage in the rat

Excessive activation of the N-methyl-D-aspartate (NMDA) subtype of glutamate receptor has been implicated in the sequence of neurochemical events that results in irreversible neuronal damage in cerebral ischemia. The effects of the NMDA antagonist (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate (MK-801) upon the amount of ischemic brain damage has been assessed quantitatively in the lightly anesthetized rat. Focal cerebral ischemia was produced by the permanent occlusion of one middle cerebral artery (MCA), and the animals were killed 3 hours after the arterial occlusion. MK-801 (0.5 mg/kg) was administered intravenously either 30 minutes prior to MCA occlusion or 30 minutes after the induction of ischemia. Pretreatment with MK-801 reduced the volume of ischemic damage both in the cerebral cortex (by 38% compared with untreated rats with MCA occlusion; p less than 0.01) and in the caudate nucleus (by 18% compared with controls; p less than 0.05). Treatment with MK-801, initiated 30 minutes after MCA occlusion, reduced the volume of ischemic damage in the cerebral cortex (by 52% compared with controls; p less than 0.01). The volume of ischemic damage in the caudate nucleus was minimally influenced by MK-801 treatment initiated after MCA occlusion. The antiischemic effects of MK-801 were readily demonstrable despite the hypotension that MK-801 induced in rats anesthetized with halothane (0.5%), nitrous oxide (70%), and oxygen (30%). The potency of MK-801 in reducing ischemic brain damage, even when administered after the induction of ischemia, highlights the potential use of NMDA receptor antagonists for the treatment of focal cerebral ischemia in humans.     

Acute cerebral infarction and changes of regional cerebral blow flow (rCBF) following experimental middle cerebral artery (MCA) occlusion

An unanaesthetized cat model of acute focal cerebral ischaemia has been established by the technique of transorbital snare ligature for middle cerebral artery (MCA) occlusion. The model was used to investigate the patterns of changes of regional cerebral blood flow (rCBF) for up to 16 h following MCA occlusion by the hydrogen clearance technique and to explore the correlation among changes of rCBF, neurological deficits, and pathological changes. The animals were divided into 2 groups according to the size of infarct which was identified by 2% triphenyl tetrazolium chloride (TTC) solution stain. Infarct larger than 10% of coronal section surface of ipsilateral hemisphere was found in 18 cats (Group A), and a smaller infarct was found in the remaining 7 cats (Group B). Between these 2 groups, there was a statistically significant difference in the average rCBF value of the ipsilateral MCA territory. The increasing grade of contralateral paralysis correlated well with the decreasing rCBF in the ipsilateral MCA territory, and the increasing grade of cerebral hemispheric swelling was directly proportional to the increasing grade of paralysis. Three patterns of changes of rCBF were observed: persistent severe ischaemia (15 cats, Group A); persistent mild to moderate ischaemia (7 cats, Group B); and immediate severe ischaemia followed by early post-ischaemic hyperaemia and death (3 cats, Group A). During the initial stage of ischaemia, a significant decrease in rCBF of the contralateral hemisphere was observed in both groups, however, in Group A rCBF gradually increased to preocclusion level, while Group B presented a further decrease in rCBF suggesting the occurrence of interhemispheric diaschisis. Thus, Group B appeared to take advantage of diaschisis during the late phase of infarct development, as well as the substantial collateral flow from the surrounding anterior and posterior cerebral artery territories.     

Cortical spreading depression causes a long-lasting decrease in cerebral blood flow and induces tolerance to permanent focal ischemia in rat brain.

Cortical spreading depression (CSD) has previously been shown to induce tolerance to a subsequent episode of transient cerebral ischemia. The objective of the present study was to determine whether CSD also induces tolerance to permanent focal ischemia and, if so, whether tolerance may be mediated by alterations in cerebral blood flow (CBF). Sprague-Dawley rats were preconditioned by applying potassium chloride to one hemisphere for 2 hours, evoking 19 +/- 5 episodes of CSD (mean +/- SD, n = 19). Three days later, the middle cerebral artery (MCA) was permanently occluded using an intraluminal suture. In a subset of animals, laser Doppler blood flow (LDF) was monitored over the parietal cortex before and during the first 2 hours of MCA occlusion. Preconditioning with CSD reduced the hemispheric volume of infarction from 248 +/- 115 mm3 (n = 18) in sham-conditioned animals to 161 +/- 81 mm3 (n = 19, P< 0.02). Similarly, CSD reduced the neocortical volume of infarction from 126 +/- 82 mm3 to 60 +/- 61 mm3 (P < 0.01). Moreover, preconditioning with CSD significantly improved LDF during MCA occlusion from 21% +/- 7% (n = 9) of preischemic baseline in sham-conditioned animals to 29% +/- 9% (n = 7, P< 0.02). Preconditioning with CSD therefore preserved relative levels of CBF during focal ischemia and reduced the extent of infarction resulting from permanent MCA occlusion. To determine whether CSD may have altered preischemic baseline CBF, [14 C]iodoantipyrine was used in additional animals to measure CBF 3 days after CSD conditioning or sham conditioning. CSD, but not sham conditioning, significantly reduced baseline CBF in the ipsilateral neocortex to values 67% to 75% of those in the contralateral cortex. Therefore, CSD causes a long-lasting decrease in baseline CBF that is most likely related to a reduction in metabolic rate. A reduction in the rate of metabolism may contribute to the induction of tolerance to ischemia after preconditioning with CSD.     

Impairment of Fos protein formation in the rat infarct borderzone by MK-801, but not by NBQX

In the present immunocytochemical study, we investigated the mechanism of Fos protein induction and the regional distribution of the Fos protein in brains of spontaneously hypertensive rats subjected to 2 h of permanent middle cerebral artery occlusion (MCAO). Rats were administered either saline or a glutamate receptor antagonist; the non-competitive NMDA receptor antagonist MK-801 or the AMPA receptor antagonist NBQX which are known to be able to reduce infarct size in MCA occluded rats. The saline treated rats showed presence of Fos protein in nerve cell nuclei throughout the cortical and striatal infarct borderzone, but no staining in the infarct core or contralateral hemisphere. MK-801 almost totally abolished this expression of Fos protein whereas NBQX had no significant effect on Fos protein expression. It is suggested that the Fos protein induction is due to repeated spreading depressions mediated by NMDA receptors in the infarct borderzone, and that Fos protein due to its persistence in the tissue can be used as a histochemical marker of borderzone tissue at risk for eventually becoming recruited in the infarct.     

Neuropeptide Y and the cerebral circulation

The significance of neuropeptide Y (NPY) in the cerebral circulation has been examined in the rat using immunocytochemistry, isolated cerebral artery preparations, and quantitative autoradiographic techniques for determining local CBF and glucose utilisation. In the rat the middle cerebral artery and the lenticulostriate artery from which blood is supplied to the caudate nucleus were found to be invested with numerous perivascular NPY immunoreactive nerve fibres. NPY (3-300 nM) contracted rat middle cerebral artery segments in a concentration-dependent manner. Intracerebral microinjections of NPY (200 pmol) or vehicle (1 microliter) were performed in rats after full recovery from anaesthesia via previously implanted guide cannulae. Following injection of NPY into the striatum, local blood flow was markedly decreased by 36% throughout the ipsilateral caudate nucleus (e.g., from 104 +/- 25 to 67 +/- 15 ml 100 g-1 min-1; mean +/- SD), whereas glucose use in this region was not altered significantly (e.g., 73 +/- 8 and 74 +/- 10 mumol 100 g-1 min-1 with vehicle and NPY, respectively). Intrastriatal NPY did not alter CBF or glucose use in the majority of other brain areas, including all of the 40 contralateral regions examined and almost all regions within the ipsilateral hemisphere. In a small number of highly discrete brain areas remote from the injection site (e.g., amygdala), there were significant reductions in blood flow with minimal changes in glucose use. Since NPY is present around rat cerebral blood vessels, is capable of evoking their contraction, and has the ability to produce reductions in blood flow independently of oxidative metabolism, this neuropeptide may be of major importance in cerebrovascular regulation.