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R Prado

Publications and source records attributed to R Prado.

At least 37 records · Page 2Linked to original sources
PubMed1996

Cerebral protection by intermittent reperfusion during temporary focal ischemia in the rat.

Temporary arterial occlusion has been routinely used as an adjunct in intracranial aneurysm surgery. This has commonly been performed using a protocol of multiple short periods of occlusion alternating with periods of restoration of normal circulation. Recently, the logical basis of this method has come under scrutiny. There is extensive experimental evidence to suggest that repetitive, brief periods of global ischemia may cause more severe cerebral injury than an equivalent single period of global ischemia. Only recently has this issue begun to be addressed with regard to focal ischemia. Hence, despite the common use of temporary clipping, little experimental data are available regarding the ischemic consequences of temporary arterial occlusion with periods of reperfusion versus uninterrupted temporary occlusion. To investigate this issue, a protocol of occlusion/reperfusion that simulates the temporal profile that occurs during surgery was performed in a rat model of focal ischemia. Sixteen anesthetized Sprague-Dawley rats were divided into two groups. The animals in Group I underwent 60 minutes of uninterrupted middle cerebral artery occlusion and the animals in Group II were subjected to six separate 10-minute occlusion periods with 5 minutes of reperfusion between occlusions. Histopathological analysis was performed 72 hours postischemia. Group I had significantly increased mean infarction volumes (50.0 +/- 12.1 mm3) compared to Group II (8.7 +/- 3.1 mm3) (p = 0.008). Injuries in Group I occurred in both the cortex and striatum, whereas Group II showed only striatal injuries. Furthermore, the extent of the injuries in Group II was less severe, characterized by ischemic neuronal injury rather than frank infarction. The results indicate that intermittent reperfusion is neuroprotective during temporary focal ischemia and support the hypothesis that intermittent reperfusion is beneficial if temporary clipping is required during aneurysm repair.

Animals↗
PubMed1995

Ischemia-induced changes in extracellular levels of striatal cyclic GMP: role of nitric oxide.

Using microdialysis, we evaluated temporal changes in striatal extracellular cGMP level following ischemia and its relationship to nitric oxide (NO) production. In untreated animals, significant elevation of cGMP was observed during ischemia and during 4 h of recirculation. In animals treated with L-NAME ischemia induced a modest increase in the cGMP level, but this level was significantly lower than that observed in the untreated animals. These results demonstrate first, that the microdialysis technique can be used to detect changes in extracellular cGMP levels during ischemia and second, that ischemia and recirculation induce a rise in cGMP which is diminished by nitric oxide synthase inhibition, suggesting a linkage to NO production.

Analysis of Variance↗
PubMed1995

A dual role for nitric oxide in NMDA-mediated toxicity in vivo.

Nitric oxide has been implicated in N-methyl-D-aspartate (NMDA)-mediated damage in vitro; however, its role in excitotoxic damage in vivo is not clear. In the present study we evaluated the histopathological and hemodynamic consequences of intrastriatal injections of various doses of NMDA and determined the effects of nitric oxide synthase inhibition on these changes. NMDA was injected into the striatum at doses of 50, 150, and 300 nmol with or without N omega-nitro-L-arginine methyl ester (L-NAME; 100 micrograms, locally). Three days following injections histopathological assessment was performed by morphometric analysis of the lesion area in multiple sections taken from the anterior to the posterior borders of the lesion. In animals injected with 150 and 300 nmol of NMDA (+/- L-NAME), local CBF (lCBF) was determined 30 min following injections using 14C-iodoantipyrine autoradiography. All NMDA-treated animals showed a well-demarcated lesion extending beyond the injection site. The volume of the lesion correlated significantly with the NMDA dose injected. The effects of L-NAME on lesion size were dependent on the dose of the NMDA. The lesion induced by 50 nmol of NMDA was not affected by L-NAME. With a dose of 150 nmol of NMDA, L-NAME induced a 43% increase in lesion volume. In contrast, a 38% decrease in lesion size was observed in animals treated with 300 nmol of NMDA combined with L-NAME. At a dose of 150 nmol, NMDA induced a significant elevation in lCBF, which was restricted to regions close to the injection site including the center areas of the anterior and middle striatum. The increase in lCBF observed with 150 nmol of NMDA was significantly attenuated in the NMDA + L-NAME-treated group. The lCBF changes induced by 300 nmol of NMDA were not significantly different from those in the 150-nmol group; however, the extent of the regions involved was larger. The increases in lCBF were observed in all striatal regions including the central and peripheral areas. L-NAME did not have a significant effect on the lCBF changes induced by NMDA at a dose of 300 nmol. These data suggest that in vivo the involvement of nitric oxide in NMDA toxicity depends on the NMDA dose and on the participation of hemodynamic mechanisms secondary to NMDA exposure.

Animals↗
PubMed1995

Acadesine reduces indium-labeled platelet deposition after photothrombosis of the common carotid artery in rats.

BACKGROUND AND PURPOSE: The adenosine-regulating agent acadesine has been shown to reduce the incidence of myocardial infarction and stroke after cardiopulmonary bypass surgery. The present study examined the effect of acadesine on the accumulation of indium-labeled platelet emboli and infarct size after photothrombosis of the common carotid artery. METHODS: Rats were anesthetized with halothane and preloaded with 111In-tropolone-labeled platelets (50 to 80 microCi) 30 minutes before nonocclusive common carotid artery thrombosis induced by a rose bengal-mediated photochemical insult. Intravenous infusion of acadesine (0.5, 1, or 2 mg/kg per minute) or vehicle was begun 30 minutes before right common carotid artery thrombosis and continued for an additional 15 minutes. Rats were then killed and brains processed for the autoradiographic quantitation of labeled platelet aggregates. In a separate group of rats, infarct areas and volumes were determined in treated (acadesine 1 mg/kg per minute) (n = 9) and nontreated (n = 9) rats 7 days after thrombosis. RESULTS: Although the ratio of right-to-left common carotid artery radioactivity was not affected by treatment, acadesine at 1 and 2 mg/kg per minute significantly decreased (P < .01) platelet deposition within the right cerebral cortex, hippocampus, and striatum. For example, within the frontoparietal cortex, numbers of platelet aggregates were 11.8 +/- 1.8 (mean +/- SEM), 6.1 +/- 1.4, 2.3 +/- 0.6, and 3.2 +/- 0.8 in rats infused with vehicle, 0.5, 1, and 2 mg/kg per minute acadesine, respectively. In addition, infarct volume was reduced by 48% in acadesine-treated (1 mg/kg per minute) rats, with a significant reduction in infarct area at the coronal level 3.7 mm anterior to bregma (P < .01). CONCLUSIONS: These results support a prophylactic role for acadesine in reducing the accumulation of platelet emboli during vascular thrombosis and subsequent brain infarction. Acadesine treatment in patients at risk for embolic stroke could potentially lead to cerebral protection.

Aminoimidazole Carboxamide↗
PubMed1995

Nonocclusive common carotid artery thrombosis in the rat results in reversible sensorimotor and cognitive behavioral deficits.

BACKGROUND AND PURPOSE: Microemboli released during transient ischemic attack, stroke, and cardiac surgery are thought to cause a variety of functional deficits in humans. The purpose of this study was to characterize the type and extent of neurobehavioral deficits present after photochemically induced common carotid artery thrombosis (CCAT), a thromboembolic model of stroke in the rat that results in a platelet emboli shower. METHODS: Thirty-two male Wistar rats were assigned to four groups. Groups 1 (n = 8) and 3 (n = 8) were long-term (6-week survival) and short-term (2-week survival) experimental groups subjected to right CCAT with the use of the photochemical technique. Groups 2 (n = 8) and 4 (n = 8) served as sham-operated controls for each experimental group. A battery of behavioral tests was applied daily beginning 24 hours after thrombosis; this consisted of elicited forelimb placing, postural reflex, beam balance, beam walking, and open field activity. Cognitive testing with a water maze task was performed on post-CCAT days 30 to 33 for groups 1 and 2 and on post-CCAT day 2 for groups 3 and 4. Ten-micrometer coronal brain sections were stained with hematoxylin and eosin, and infarct location and frequency were determined. RESULTS: Significant sensorimotor deficits were observed, which recovered within 2 weeks after CCAT. The data that follow are derived by combining the two experimental groups and comparing these with the two sham groups. The following tests showed significant effects after CCAT: contralateral elicited forelimb placing, ipsilateral elicited forelimb placing, beam balance, and beam walking score. Cognitive dysfunction was seen acutely (group 3 animals) at 2 days after CCAT; Morris water maze length and latency to target were significantly greater in the experimental group. No deficits were seen in postural reflex, open field activity, or delayed cognitive testing. Histopathological assessment revealed small infarcts in 11 of 16 thrombosed rats. However, a strong relationship between neurobehavioral deficits and infarct location was not consistently demonstrated. CONCLUSIONS: CCAT produces consistent sensorimotor and cognitive behavioral deficits that recover within 2 weeks of injury. Behavioral outcome was not necessarily associated with overt histopathological damage, suggesting that reversible injury mechanisms, both vascular and neuronal, may be partly responsible for the temporary loss of function. These data strengthen the role of CCAT as a clinically relevant model of thromboembolic stroke.

Animals↗
PubMed1995

A photothrombotic 'ring' model of rat stroke-in-evolution displaying putative penumbral inversion.

BACKGROUND AND PURPOSE: To facilitate reproducible and rigorous study of a tissue zone at risk of encroaching ischemic damage, we propose a new model in which the potentially compromised tissue lies within rather than perifocal to an ischemic locus. The perimeter of the "zone at risk" is defined by a photothrombotically produced cortical lesion in the shape of a toroid (or "ring"). METHODS: The exposed crania of erythrosin B-injected rats were irradiated with a 514.5-nm laser beam, configured as a 5-mm-diameter ring, to yield a ring-shaped lesion caused by photochemically induced platelet occlusion of cortical vasculature. Developing perfusion deficits in the interior region were revealed by carbon black infusion. Tissue damage and infarct volumes were assessed by light and electron microscopy, and blood-brain barrier integrity was assessed with Evans blue dye and horseradish peroxidase as tracers. RESULTS: For rats injected with 17 mg/kg erythrosin B and irradiated for 2 minutes with a ring beam intensity of 0.92 W/cm2 (beam power of 65 mW), carbon black infusion at times up to 4 hours demonstrated a shallow cortical ring lesion encircling a fully patent zone at risk, which by 24 hours evinced an essentially complete perfusion deficit. At times up to 24 hours, the ring lesion was penetrated at the pial surface by distal branches of the middle cerebral and anterior cerebral arteries. Stereotaxically based histopathological assessment showed that by 24 hours the lesion spanned the cortical thickness. Lesion volume increased from 14.5 +/- 8.0 mm3 (mean +/- SD) (n = 8) to 46.2 +/- 15.6 mm3 (n = 8) between 4 and 24 hours after irradiation (P < .01), but the anteroposterior lesion diameter did not change significantly between 4 hours (6.00 +/- 1.03 mm; n = 9) and 24 hours (6.75 +/- 1.15 mm; n = 9). CONCLUSIONS: The present model of slowly developing but inevitable cortical tissue death in a sequestered area should facilitate more precise observations of the evolution of tissue metabolic responses, from the impending onset of ischemia to the threshold of irreversible damage. This system may prove efficient for evaluating treatments intended to salvage a penumbral region.

Animals↗
PubMed1994

Glutamate antagonist MK-801 attenuates incomplete but not complete infarction in thrombotic distal middle cerebral artery occlusion in Wistar rats.

The purpose of this study was to investigate the effects of a non-competitive N-methyl-D-aspartate antagonist, MK-801, on incomplete infarction (selective neuronal necrosis), a zone of which had been found previously in a thrombotic distal middle cerebral artery (MCA) occlusion model in Wistar rats. Male Wistar rats were treated with 1 mg/kg of MK-801 or saline 30 min before MCA occlusion. Laser irradiation with intravenous administration of Rose Bengal dye was used to cause thrombotic distal MCA occlusion. The ipsilateral common carotid artery was occluded permanently and the contralateral carotid artery for 60 min. Head temperature was controlled at 36 degrees C. Cerebral blood flow (CBF) was determined with laser-Doppler flowmetry. Three days after the ischemic insult, brains were perfusion-fixed and volumes of cortical (complete and incomplete) infarction were determined. There were no significant differences in physiological variables or CBF between the two groups. Volumes of complete infarction were equivalent between the two groups (94.9 +/- 15.6 mm3 and 91.6 +/- 14.0 mm3 in the control and MK-801 treated groups, respectively). In MK-801 treated group, the volume of incomplete infarction was reduced by 44% (6.4 +/- 1.7 mm3 vs. 3.6 +/- 2.1 mm3 in control and MK-801 treated groups, respectively, P < 0.05). Although the zone responsive to MK-801 was small in this thrombotic MCA occlusion model, our present study revealed that MK-801 has a beneficial effect on the tissue zone containing selective neuronal alterations (incomplete infarction). Our results support the concept that this drug is effective in the area of less severe ischemia.

Animals↗
PubMed1993

Effects of nitric oxide synthase inhibition on cerebral blood flow following bilateral carotid artery occlusion and recirculation in the rat.

The effects of bilateral carotid artery occlusion/recirculation on cortical CBF (cCBF) were studied in rats following the intravenous administration of either the nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester hydrochloride (L-NAME; 30 mg/kg) or an equivalent volume of saline (500 microliters). Induction of bilateral carotid occlusion (BCO) in L-NAME-treated animals resulted in a reduction of cCBF to 30% of baseline. During recirculation subsequent to 20 min of BCO, cCBF in L-NAME-infused animals remained at 30% of baseline. In contrast, cCBF in saline-treated control animals returned to the original baseline level following a similar reduction to 30-40% of baseline during BCO. These results indicate that inhibition of nitric oxide generation limits normalization of regional cortical perfusion following occlusion of proximal large cerebral vessels.

Amino Acid Oxidoreductases↗
PubMed1993

Microvascular and neuronal consequences of common carotid artery thrombosis and platelet embolization in rats.

The microvascular and neuronal consequences of nonocclusive common carotid artery (CCA) thrombosis were documented in rats. Thrombosis of the CCA was produced by a rose bengal-mediated photochemical insult and regional patterns of blood-brain barrier (BBB) disruption were documented by horseradish peroxidase (HRP) histochemistry at 15 min (n = 12), 4 h (n = 3), 1 day (n = 5) or 7 days (n = 5) after vascular injury. At 15 min and 4 h after thrombosis, multiple foci of BBB disruption were present throughout the thrombosed hemisphere; protein leakage was occasionally detected contralaterally. Extravasated HRP was associated with well-perfused arterioles and arterioles containing luminal platelet aggregates at different stages of degranulation. Evidence for local platelet adhesion and aggregation or endothelial disruption at these sites was not detected. However, HRP-containing endothelial plasmalemmal vesicles were present at leaky sites. Variable degrees of parenchymal injury were documented including dendritic and astrocytic swelling with neuronal necrosis. By 1 day after CCA thrombosis, the overall frequency of permeable sites, more commonly associated with luminal leukocytes and parenchymal necrosis, was reduced. At 7 days, vessels permeable to HRP were associated with tissue necrosis, reactive astrocytes and microglial infiltration. Arteriole wall thickening and leukocyte accumulation within arterioles and venules were also detected. Widespread platelet embolization leading to variable degrees of BBB disruption and tissue injury occurs after CCA thrombosis. Acute abnormalities in vascular permeability are thus hypothesized to play an important role in the acute pathogenesis of cerebrovascular thrombosis. Delayed leukocyte accumulation in this model of embolic infarction may represent a secondary insult to the injured brain.

Animals↗
PubMed1993

Selective brain cooling increases cortical cerebral blood flow in rats.

To evaluate the effect of selective brain cooling on cortical cerebral blood flow, we reduced brain temperature in nitrous oxide anesthetized adult rats using a high speed fan while keeping rectal temperature at 37-38 degrees C. During selective brain cooling, cortical cerebral blood flow, as measured by laser-Doppler flowmetry, increased to 215 +/- 26% (mean +/- SE) of baseline at a cortical brain temperature of 30.9 +/- 0.5 degrees C and a rectal temperature of 37.5 +/- 0.1 degrees C. During rewarming, as brain temperature increased, cortical cerebral blood flow decreased. The cerebral vasodilatory response to hypothermia may explain its protective effects during and after cerebral ischemia.

Animals↗
PubMed1993

Transient platelet accumulation in the rat brain after common carotid artery thrombosis. An 111In-labeled platelet study.

BACKGROUND AND PURPOSE: Thromboembolic events are a major cause of ischemic stroke. To obtain evidence for platelet embolization after cerebrovascular injury, the accumulation of indium-labeled platelets was documented after photothrombosis of the rat common carotid artery. METHODS: Heterologous blood was collected from donor rats, and the isolated platelets were labeled with 111In-tropolone. Labeled platelets were then infused into Wistar rats 30 minutes before right carotid artery thrombosis. Nonocclusive common carotid artery thrombosis was induced by a laser-driven rose bengal-mediated photochemical insult to the vascular endothelium, and the rats were killed 15 minutes or 3 hours later. Carotid arteries and brains were immediately removed and dissected for regional radioactivity assessment or sectioned for the autoradiographic visualization of platelet emboli. RESULTS: At 15 minutes after thrombosis, the ratio of right-to-left common carotid artery radioactivity was significantly elevated compared with control (33 +/- 12 [mean +/- SEM] versus 0.97 +/- 0.2). Within individual brain regions, including the frontal and frontoparietal cortices and hippocampus, significant elevations in right-to-left radioactivity ratios were also documented. Autoradiographic images revealed multiple foci of 111In-labeled platelets throughout the thrombosed hemisphere. At the level of the frontal cortex, bilateral platelet accumulation was seen. Regional counts demonstrated significantly increased platelet density within selective cortical and subcortical regions. In contrast to the 15-minute findings, right-to-left ratios of carotid arteries or brain regional radioactivities were not significantly elevated at 3 hours after injury. In addition, the areal densities of autoradiographically visualized platelets in the 3-hour group were not different from control except in the right frontal cortex. CONCLUSIONS: These data demonstrate (1) the acute accumulation of labeled platelets in downstream vessels after nonocclusive common carotid artery thrombosis, (2) that platelet accumulation is widespread and also involves contralateral areas, and (3) that platelet accumulation within the thrombosed carotid artery and brain is largely transient.

Animals↗
PubMed1993

Comparative histopathologic consequences of photothrombotic occlusion of the distal middle cerebral artery in Sprague-Dawley and Wistar rats.

BACKGROUND AND PURPOSE: We have developed a minimally invasive model of photothrombotic occlusion of the distal middle cerebral artery in rats and have evaluated the patterns and features of the resulting histopathologic injury in two normotensive strains. METHODS: Food-deprived male Sprague-Dawley (n = 14) and Wistar (n = 10) rats anesthetized with halothane/nitrous oxide underwent a small craniotomy to expose the right distal middle cerebral artery just above the rhinal fissure. The animals were injected intravenously with the photosensitizing dye rose bengal, and the distal middle cerebral artery was irradiated with light from an argon laser-activated dye laser at three separate points to induce thrombotic occlusion. The ipsilateral common carotid artery was then permanently occluded, and the contralateral common carotid artery was occluded for 60 minutes. Three days later, the brains were perfusion-fixed and prepared for histopathologic examination, and infarct volume was determined by quantitative planimetry. RESULTS: In Sprague-Dawley rats, a large consistent temporoparietal cortical infarct was observed; mean +/- SD infarct volume was 130.5 +/- 40.0 mm3 (coefficient of variation, 30.7%) and a relatively small adjacent zone of selective neuronal necrosis ("incomplete infarction"), amounting to only 9.1% of the total injury volume, was also seen. By contrast, Wistar rats had smaller and more variable cortical infarcts (volume, 48.4 +/- 26.9 mm3; coefficient of variation, 55.6%) but displayed a much more substantial zone of incomplete cortical infarction (volume, 20.8 +/- 10.1 mm3; 30.1% of the total injury volume). In neither strain was infarct size related to alterations of blood pressure. In both strains, infarcts were limited to the cortex, typically involving the parietal cortex, somatosensory cortex, and forelimb region. Three rats exhibited infarcts in the contralateral hemisphere. CONCLUSIONS: This model has the advantages of necessitating only minimal surgery, allowing the dura to remain intact, and avoiding mechanical trauma to the brain surface. In Sprague-Dawley rats, the resulting large cortical infarct exhibited relatively small interanimal variation, making the model suitable, for example, for replicate studies of pharmacotherapy. In Wistar rats, the large zone of incomplete infarction, a unique feature heretofore undescribed in rodent models of permanent focal ischemia, lends the model to the study of the pathomechanisms underlying graded cortical ischemic injury.

Animals↗
PubMed1993

Failure of MK-801 to reduce infarct volume in thrombotic middle cerebral artery occlusion in rats.

BACKGROUND AND PURPOSE: We examined the effects of the noncompetitive N-methyl-D-aspartate receptor antagonist MK-801 using a newly developed stroke model of thrombotic distal middle cerebral artery occlusion under conditions of carefully controlled head temperature. METHODS: Male Sprague-Dawley rats were treated with 1 mg/kg of MK-801 or saline before the induction of ischemia. An argon laser-activated dye laser (562 nm) was used to cause thrombotic distal middle cerebral artery occlusion. In experiments 1 and 2, the single laser beam (20 mW) was separated into three beams. Each beam was positioned onto the distal middle cerebral artery at three sites along the vessel. The photosensitizing dye rose bengal (20 mg/kg) was administered intravenously over 2 minutes; the three points were then irradiated for 3 minutes. In experiment 3, higher power of the laser (three separate irradiations using a single beam of 20 mW) was used. The ipsilateral common carotid artery was occluded permanently, and the contralateral carotid artery was occluded for 60 minutes. Head temperature was controlled at 36 degrees C in experiment 1 and not controlled in experiments 2 and 3. Three days after the ischemic insult, brains were perfusion-fixed and infarct volumes were determined. RESULTS: Head temperature was mildly hypothermic (34-35 degrees C before ischemia, with a further decrease of 1-2 degrees C during the initial 60 minutes of ischemia) in experiment 2. However, no differences were observed in head temperature between the MK-801-treated and control groups. Cortical infarct volume in experiment 1 was 89 +/- 29 mm3 (mean +/- SD) in the treated group, which was not different from the control value of 84 +/- 40 mm3. Infarct volumes were smaller (58 +/- 35 mm3 and 54 +/- 14 mm3) in the control groups of experiments 2 and 3, respectively. However, MK-801 also failed to reduce infarct volumes in experiments 2 and 3. CONCLUSIONS: MK-801 is not effective in this stroke model of focal thrombotic infarction under conditions of either controlled (normothermic) or uncontrolled (mildly hypothermic) head temperature.

Animals↗
PubMed1992

Ischemia-induced changes in extracellular levels of striatal cyclic AMP: role of dopamine neurotransmission.

Dopamine has been demonstrated to be involved in the development of ischemic neuronal damage in the striatum. This detrimental effect of dopamine may involve activation of second messenger systems, such as the cyclic AMP (cAMP) cascade, which may enhance the susceptibility of striatal neurons to ischemia. In the present study, we have evaluated the relationship between ischemia-induced changes in cAMP and dopamine neurotransmission. Microdialysis probes were implanted in both striata, and a D1 antagonist (SCH-23390, 100 microM) was administered through one probe and modified Ringer's solution through the other. After a stabilization period, rats (n = 6) were subjected to 20 min of ischemia by two-vessel occlusion plus hypotension. Extracellular samples were collected from both striata, before, during, and after ischemia, and analyzed for cAMP by radioimmunoassay. Ischemia induced a significant increase in extracellular cAMP (means +/- SE, fmol/microliter; baseline: 4.35 +/- 1.1, ischemia: 12.2 +/- 1.98), which was also observed at 4 h of recirculation (mean level of 8.45 +/- 1.14). Treatment with the D1 antagonist significantly inhibited the rise in extracellular cAMP during ischemia and recirculation. These results indicate that an ischemia-induced surge in dopamine and activation of D1 receptors are involved in the generation of cAMP during ischemia and recirculation. Because activation of the adenylate cyclase cascade may modulate the effects of glutamate, generation of cAMP through this pathway may play a role in facilitating the injurious effects of dopamine during ischemia.

Animals↗
PubMed1992

Serotonin release into plasma during common carotid artery thrombosis in rats.

BACKGROUND AND PURPOSE: We have tested the hypothesis that platelet-derived serotonin is released into the bloodstream during cerebrovascular thrombosis. METHODS: Nonocclusive common carotid artery thrombosis was produced photochemically in 22 anesthetized adult male Wistar rats using the photosensitizing dye rose bengal and irradiation with an argon-pumped dye laser. Plasma serotonin levels were recorded before, during, and after photothrombosis by intra-arterial in vivo microdialysis with the probe placed distal to the site of thrombosis. RESULTS: During and immediately after the common carotid artery thrombosis, serotonin levels increased significantly to a peak value of 781 nmol/l (p less than 0.001 by analysis of variance), representing a 15-fold increase compared with baseline levels. The increased serotonin levels gradually decreased but remained significantly elevated for 90 minutes. Ultrastructural analysis of the carotid thrombi identified a dense mass of aggregated platelets at various stages of degranulation. CONCLUSIONS: These results are the first to demonstrate directly that serotonin accumulation occurs in plasma during and after the acute phase of common carotid artery thrombosis. Increased plasma serotonin levels may play a major role in the cerebral blood flow and blood-brain barrier abnormalities previously documented in this model of large-vessel thrombotic stroke.

Animals↗
PubMed1992

Endothelium-derived nitric oxide synthase inhibition. Effects on cerebral blood flow, pial artery diameter, and vascular morphology in rats.

BACKGROUND AND PURPOSE: We determined the effects of inhibiting the production of cerebral endothelium-derived nitric oxide on pial artery diameter, cortical blood flow, and vascular morphology. METHODS: An inhibitor of endothelium-derived nitric oxide synthesis, NG-nitro-L-arginine methyl ester hydrochloride (L-NAME), or an equivalent volume of 0.9% saline was infused into rats intra-arterially in a retrograde fashion via the right external carotid artery at a rate of 3 mg/kg/min to a total dose of 190 mg/kg or intravenously at 1 mg/kg/min to a total dose of 15 mg/kg. Large pial arteries were continuously visualized through an operating microscope, and cortical cerebral blood flow was monitored by laser-Doppler flowmetry. To localize areas of morphological interest, the protein tracer horseradish peroxidase was injected 15 minutes before termination of the L-NAME infusion and the rats were perfusion-fixed 15 minutes later for light and electron microscopic analysis. RESULTS: Infusion of L-NAME significantly raised arterial blood pressure at both doses (for 190 mg/kg, from 103.2 +/- 3.4 to 135 +/- 3.4 mm Hg; for 15 mg/kg, from 125 +/- 2.8 to 144.4 +/- 4.0 mm Hg). Pial arteries constricted within 10 minutes after the start of the intracarotid infusion to 40% of the preinfusion diameter, while cortical cerebral blood flow decreased to an average of 72.5% of that at baseline. Morphological abnormalities in the experimental rats included microvascular stasis and focal areas of blood-brain barrier disruption to protein. Ultrastructural examination of cortical leaky sites revealed constricted arterioles with many endothelial pinocytotic vesicles and microvilli. CONCLUSIONS: These observations suggest that inhibition of endothelium-derived nitric oxide synthesis affects the relation between cerebral arterial diameter and cerebral blood flow and can lead to subtle cerebral vascular pathological changes consistent with focal brain ischemia.

Amino Acid Oxidoreductases↗
PubMed1991

Hemodynamic consequences of common carotid artery thrombosis and thrombogenically activated blood in rats.

We documented the hemodynamic consequences of nonocclusive common carotid artery thrombosis (CCAT) and tested the hypothesis that vasoactive substances capable of altering local CBF (LCBF) are released into the systemic circulation following cerebrovascular injury. Ten minutes after photochemically induced CCAT, an autoradiographic determination of LCBF was conducted with [14C]iodoantipyrine. In blood transfusion studies using donor and recipient rats, a 1-ml sample of thrombogenically activated blood (TAB) collected downstream from the forming thrombus was reinjected into a recipient rat 15 or 60 min before CBF study. A heterogeneous pattern of abnormal LCBF was documented in the ipsilateral hemisphere of CCAT rats and recipient rats receiving TAB 15 min before CBF study. Acute hemodynamic abnormalities included ischemic (less than 35% of control) and hyperemic (greater than 125% of control) foci and more global reductions (50-80% of control) in cortical and subcortical LCBF. Border zone hyperemia exceeding 2.0 ml/g/min was associated with focal sites of severe LCBF reductions. Although recipient rats that received TAB 15 min before CBF study displayed similar hemodynamic abnormalities, LCBF values in 60-min recipient rats were not significantly different from control despite ischemic foci. Humoral factors generated during CCAT appear to be responsible for the acute LCBF consequences of cerebrovascular thrombosis. Vasoactive substances released from a thrombotic site, capable of regionally affecting vascular reactivity in a time-dependent fashion, might be expected to participate in the pathogenesis of transient ischemic attacks and acute stroke.

Animals↗