Unilateral Middle Cerebral Artery Occlusion Research Articles

Glial gap junctional communication involvement in hippocampal damage after middle cerebral artery occlusion

Most patients with stroke caused by middle cerebral artery occlusion (MCAO) show cognitive deficit that is generally regarded as resulting from damage to the cerebral cortex rather than the hippocampus. Whether MCAO induces hippocampal damage and whether this contributes to the cognitive defects remains unclear. Here we investigate the hippocampal damage and its correlation to cognitive defects after exclusively unilateral MCAO and the underlying mechanism for that damage. Patients were assessed for hippocampal damage by magnetic resonance imaging (MRI) and magnetic resonance angiography (MRA), and the Mini Mental-Status Evaluation (MMSE) and Rey Auditory Verbal Learning Test (RAVLT) were used to assess for cognitive defects. We provide the first evidence that patients with exclusively unilateral MCAO showed hippocampal damage characterized by an infarct-size-independent atrophy and alterations in neuronal and glial metabolites in the ipsilateral hippocampus, in parallel with cognitive impairment. Rodent MCAO also induced delayed shrinkage and pyramidal neuronal death in the ipsilateral hippocampus and an impairment of hippocampal-dependent spatial memory. Blocking Gap junctional communication (GJC) with carbenoxolone or downregulation of connexin43 (Cx43) significantly increased the survival of the pyramidal neurons in the ipsilateral hippocampus and improved behavioral scores. Furthermore, Cx43 heterozygous mice showed reduced shrinkage and metabolite abnormality in ipsilateral hippocampus after MCAO. Astroglial GJC plays a significant role in MCAO-induced remote hippocampal damage and cognitive impairment. It might be possible to improve the cognition in patients with MCAO by manipulating interastrocytic communication via the gap junction channels.

Imaging Mass Spectrometry Detection of Gangliosides Species in the Mouse Brain following Transient Focal Cerebral Ischemia and Long-Term Recovery

Gangliosides, a member of the glycosphingolipid family, are heterogeneously expressed in biological membranes and are particularly enriched within the central nervous system. Gangliosides consist of mono- or poly-sialylated oligosaccharide chains of variable lengths attached to a ceramide unit and are found to be intimately involved in brain disease development. The purpose of this study is to examine the spatial profile of ganglioside species using matrix-assisted laser desorption/ionization (MALDI) imaging (IMS) following middle cerebral artery occlusion (MCAO) reperfusion injury in the mouse. IMS is a powerful method to not only discriminate gangliosides by their oligosaccharide components, but also by their carbon length within their sphingosine base. Mice were subjected to a 30 min unilateral MCAO followed by long-term survival (up to 28 days of reperfusion). Brain sections were sprayed with the matrix 5-Chloro-2-mercaptobenzothiazole, scanned and analyzed for a series of ganglioside molecules using an Applied Biosystems 4800 MALDI TOF/TOF. Traditional histological and immunofluorescence techniques were performed to assess brain tissue damage and verification of the expression of gangliosides of interest. Results revealed a unique anatomical profile of GM1, GD1 and GT1b (d18∶1, d20∶1 as well as other members of the glycosphingolipid family). There was marked variability in the ratio of expression between ipsilateral and contralateral cortices for the various detected ganglioside species following MCAO-reperfusion injury. Most interestingly, MCAO resulted in the transient induction of both GM2 and GM3 signals within the ipsilateral hemisphere; at the border of the infarcted tissue. Taken together, the data suggest that brain region specific expression of gangliosides, particularly with respect to hydrocarbon length, may play a role in neuronal responses to injury.

Leptomeningeal collateral and cerebral hemodynamics in patients with ICA and MCA steno-occlusion

<title/>Objectives: The purpose of our study was to evaluate the correlation between fluid-attenuated inversion recovery (FLAIR) images of hyperintense vessel (HV), a representative of leptomeningeal vessels (LMVs), and cerebral vascular hemodynamic status in patients with steno-occlusive disease of the internal carotid artery (ICA) and middle cerebral artery (MCA).Methods: Sixty-eight patients with unilateral ICA or MCA stenosis or occlusion were enrolled. Correlations between hyperintense vessel (HV) on FLAIR images and hemodynamic status measured by quantitative single-photon emission computed tomography data were evaluated in four groups: affected (A) and non-affected (B) hemispheres in HV-positive patients, and affected (C) and non-affected (D) hemispheres in HV-negative patients.Results: The presence of HV was most frequently seen on affected hemispheres in patients with MCA and ICA occlusions. HV was significantly higher in the anterior communicating artery (AcomA) (−), posterior communicating artery (PcomA) (−) group than in other groups, which included either or both AcomA and PcomA (P = 0·008). The presence of HV had a significant correlation with transdural anastomosis (P = 0·046) and LMV (P = 1·64×10−9). No significant difference was observed in resting regional cerebral blood flow among the four groups. Vascular reserve in group A was significantly less compared to other groups (P = 0·021).Conclusions: HV is related to the hemisphere with severe ICA and MCA steno-occlusion when the circle of Willis is insufficient. The HV-positive hemisphere on the steno-occlusive side shows a lower vascular reserve than that in the contralateral hemisphere or HV-negative hemispheres.

Mechanism of Moyamoya Vessels Secondary to Intracranial Atherosclerotic Disease: Angiographic Findings in Patients with Middle Cerebral Artery Occlusion

Moyamoya vessels, which form a fragile collateral vascular network at the base of the brain, are associated with moyamoya disease and moyamoya syndrome. The mechanisms involved in their development remain unknown. This study evaluated the angiographic findings of the collateral vasculature in patients with moyamoya vessels secondary to atherosclerotic middle cerebral artery (MCA) occlusion. The study population comprised 26 patients with chronic atherosclerotic unilateral MCA occlusion who underwent digital subtraction angiography. We evaluated the presence of moyamoya vessels and intracranial arterial lesions and the degree of leptomeningeal anastomosis. We divided the patients into two groups, those with and those without moyamoya vessels, and compared clinical backgrounds and angiographic findings between the groups. Of the 26 patients, 17 had moyamoya vessels. The presence of moyamoya vessels was associated with ipsilateral or contralateral anterior cerebral artery stenosis to the occluded MCA (P = .004) and poor development of anterior leptomeningeal anastomosis (P = .012). It also was associated with vascular lesions involving more than one intracranial branch vessel in patients with moyamoya syndrome secondary to atherosclerosis. Our findings suggest that moyamoya vessels might be compensatory collateral vessels associated with poorly developed leptomeningeal collateral vessels.

Bone Marrow-Derived Nonreactive Astrocytes in the Mouse Brain After Permanent Middle Cerebral Artery Occlusion

We studied the effect of permanent unilateral middle cerebral artery occlusion (PMCAO) on the generation of bone marrow (BM)-derived astrocytes in female mice previously transplanted with enhanced green fluorescent protein-expressing BM from male donors. In addition to an untreated PMCAO group, one group of mice also received intracerebral infusion of transforming growth factor-alpha, resulting in a decrease in the size of the infarct. Two months after PMCAO, we found a specific type of astrocyte of BM origin in the side of the injury, near the lesion. These astrocytes did not express glial fibrillary acidic protein (GFAP) by conventional fluorescence immunostaining; however, GFAP was easily detectable by tyramide signal amplification. These cells also expressed S100β, confirming their astrocytic character. Unlike the endogenous reactive astrocytes, these BM-derived astrocytes did not proliferate during the first week of ischemia and did not contribute to the glial scar formation. Transforming growth factor-alpha infusion increased the number of BM-derived astrocytes, without affecting their distribution. Interestingly, exclusively by tyramide signal amplification staining, we found that endogenous astrocytes displaying an identical morphology were also present in control mouse and human brains. Our data demonstrate that a subpopulation of nonreactive astrocytes expressing low levels of GFAP can originate from transplanted BM in the ischemic brain. We believe that these cells represent a subpopulation of astrocytes earlier considered to be GFAP negative. The high number of astrocytes with identical morphology and chemical character in control brains suggest that these type of astrocytes may have important functional role in the central nervous system that calls for further studies.

Local hemodynamics dictate long-term dendritic plasticity in peri-infarct cortex.

Changes in dendritic spine turnover are a major mechanism of experience-dependent plasticity in the adult neocortex. Dendritic spine plasticity may also contribute to functional recovery after stroke, but in that setting its expression may be complicated by alterations in local tissue perfusion, especially around the infarct. Using adult Thy-1 GFP-M mice, we simultaneously recorded long-term spine dynamics in apical dendrites from layer 5 pyramidal cells and blood flow from surrounding capillaries with in vivo two-photon microscopy in peri-infarct cortex before and after unilateral middle cerebral artery occlusion. Blood flow in peri-infarct cortex decreased significantly immediately after stroke and improved gradually over time, in a distance-dependent manner from the epicenter of the infarct. However, local tissue perfusion was never fully restored even after a 3 month recovery period. On average, surviving layer 5 pyramidal neurons experienced a ∼20% decrease in spine density acutely after stroke but eventually recovered. The dynamics of this improvement were different depending on the degree of tissue perfusion acutely after arterial occlusion. Cells in ischemic areas closer to the infarct returned to normal spine density levels slowly by retaining spines, while cells in more remote regions with preserved blood flow recovered faster by adding more spines, eventually surpassing baseline spine density by 15%. Our data suggest that maintaining tissue perfusion in the area surrounding the infarct could hasten or augment synaptic plasticity and functional recovery after stroke.

Soy protein diet increases skilled forelimb reaching function after stroke in rats

Stroke is a leading cause of lasting disability. Dietary strategies aimed at increasing post-stroke outcomes are lifestyle alterations which could be easily implemented by people at risk of occlusive stroke. Soy diets have been demonstrated to provide some benefits in the short term following stroke, but longer time periods have not been studied. Further, carefully defined diets containing soy protein isolates have not been investigated. In the current study, male Long Evans Hooded rats were fed semi-purified diets containing either sodium caseinate or soy protein isolate. Rats were trained to perform the skilled forelimb reaching task and subsequently underwent unilateral middle cerebral artery occlusion (MCAO) to induce a stroke lesion. After stroke, rats remained on the same diet and were tested daily for a period of 8 weeks to observe their performance on the skilled forelimb reaching task. In the first week following stroke, rats receiving the soy protein-containing diet (SP) demonstrated less severe reaching deficits than rats fed the Na caseinate-containing diet (CAS) ( p < 0.05). These results suggest that a soy protein-based diet provides significant protection from neurological damage following MCAO stroke in rats.

Pre-ischemic exercise preserves cerebral blood flow during reperfusion in stroke

Introduction: The neuroprotective nature of exercise has been well established and the mechanisms of this protection are still a subject of much research. This study aims to determine if cerebral blood flow is constituently higher during the ischemia or reperfusion events in stroke.Materials and methods: Adult male Sprague–Dawley rats were randomly assigned into exercise or non-exercise (control) groups. Exercised rats underwent 30 minutes of running on a treadmill for 3 weeks. A 2 hour unilateral middle cerebral artery occlusion using an intraluminal filament was performed to induce ischemic stroke, followed by a 24 hour reperfusion. A sham control without exercise and middle cerebral artery occlusion was used. Laser Doppler flowmetry (LDF) and 15O-H2O positron emission tomography (PET) were used to determine cerebral blood flow, respectively. 18F-fluorodeoxy-D-glucose was used to determine cerebral metabolism in some animals. Histological analysis determined infarct volume in the same animal after blood flow examination.Results: LDF and PET both indicated that middle cerebral artery occlusion significantly (p<0·05) reduced cerebral blood flow during ischemia and reperfusion in association with reduced cerebral metabolism after stroke. However, pre-ischemic exercise significantly (p<0·05) improved cerebral blood flow during reperfusion, although cerebral blood flow remained at a similar level to that of the non-exercise stroke group during the middle cerebral artery occlusion. This improved cerebral blood flow during reperfusion was associated with decreased brain infarct volume.Conclusions: This study revealed that pre-ischemic exercise in rats improved cerebral blood flow during reperfusion, suggesting that exercise provides neuroprotection by partially ameliorating the 'no reflow' phenomenon in stroke.

Transient and bilateral increase in Neuropilin-1, Fer kinase and collapsin response mediator proteins within membrane rafts following unilateral occlusion of the middle cerebral artery in mouse

Membrane rafts, rich in sphingolipids and cholesterol, are membrane microdomains important in neuronal domain-specific signaling events such as during axonal outgrowth and neuronal death. The present study seeks to determine the spatiotemporal association of several axonal guidance signaling molecules with membrane rafts. These molecules are Neuropilin-1 (NRP-1), Fer Kinase, and collapsin response mediator proteins (CRMPs), which are known to have important functions in axonal outgrowth and neuronal death caused by cerebral ischemia. Mice were subjected to sham or a 1 h unilateral middle cerebral artery occlusion (MCAO) followed by a time course of reperfusion up to 24 h. Brain cortices were separated and membrane rafts were extracted based on their insolubility in Triton X-100 and separation by sucrose gradient fractionation. We demonstrate the early and transient induction of NRP-1 and CRMP-2 in membrane rafts in both ipsilateral and contralateral hemispheres, in contrast to an early, but sustained elevation of Fer kinase and other CRMPs (1, 3, 4, 5) in response to unilateral MCAO. The fact that NRP1/Fer kinase/CRMP-2 co-localize in membrane rafts early during ischemic injury suggests that the membrane rafts may form a scaffold to support and initiate NRP1/Fer/CRMP-2-mediated signal transduction in neuronal damage response during ischemia-reperfusion. Further understanding of the time-specific and membrane domain-specific protein–protein interaction may lead to the identification of therapeutic targets for stroke.

Effects of a high soy protein diet on forelimb function after stroke in rats

Dietary strategies aimed at increasing post‐stroke outcomes are lifestyle alterations which could be easily implemented by people at risk of stroke. Soy diets have been demonstrated to provide some benefits in the short term following stroke, but longer time periods have not been studied. Further, carefully defined diets containing soy protein isolates have not been investigated. In the current study, male LE Hooded rats were fed semi‐ purified diets containing either sodium caseinate or a soy protein isolate. Rats were trained to perform the skilled forelimb reaching task and subsequently underwent unilateral middle cerebral artery occlusion (MCAO) to induce a stroke lesion. After stroke, rats remained on the same diet and were tested daily for a period of 8 weeks to observe their performance on the skilled forelimb reaching task. In the first and second weeks following stroke, rats which were fed the soy protein diet demonstrated less severe reaching deficits than rats fed the casein diet (p&lt;0.05). Further, rats fed the soy protein diet reached statistical recovery to within baseline range at three weeks post‐stroke. Casein‐fed rats did not reach statistical recovery within the time course of the study. These results indicate that a diet rich in soy protein provides some protection from neurological damage following MCAO stroke in rats and may encourage a more rapid spontaneous recovery of skilled forelimb reaching.Grant Funding Source: none

Effects of Fentanyl Pretreatment on Regional Cerebral Blood Flow in Focal Cerebral Ischemia in Rats

Background:There are reports that opioid preconditioning induces opioid-receptor-dependent neuroprotection against cerebral ischemia. This experiment was performed to test whether pretreatment with fentanyl, a synthetic primary µ-opioid receptor agonist, would affect the regional cerebral blood flow (rCBF) in focal cerebral ischemia in rats. Methods: Twenty-four hours before permanent and unilateral middle cerebral artery (MCA) occlusion, rats were pretreated with normal saline, 200 µg/kg of fentanyl i.p. or 500 µg/kg of fentanyl i.p. The rats were anesthetized with isoflurane and were mechanically ventilated to cannulate the vessels and to occlude MCA. One hour after MCA occlusion, the rCBF was measured using ¹⁴C-iodoantipyrine. Results: The cortical rCBF decreased 1 h after MCA occlusion in all the experimental groups. In the ischemic cortex, the rCBF of the rats treated with 500 µg/kg of fentanyl was significantly greater (+80%, p < 0.05) than that of the control animals. The rCBF of the ischemic cortex of the rats treated with fentanyl 200 µg/kg seemed higher than in the control animals, but the difference was not statistically significant. The rCBF was similar in the nonischemic brain regions such as the contralateral cortex or pons among the experimental groups. Conclusion: Our data demonstrated that pretreatment with fentanyl improved the rCBF in the focal ischemic area without change in rCBF in the nonischemic cortex. Our data suggest that fentanyl could be effective in improving the rCBF in the focal ischemic area when used as a preconditioning agent and that improvement of rCBF could be one of the contributing factors of neuroprotection by opioid preconditioning.

Protective role of COMP‐Ang1 in ischemic rat brain

In cerebral ischemia, the induction of angiogenesis may represent a natural defense mechanism that enables the hypoxic brain to avoid progression into infarction. Angiopoietin-1 (Ang1) is known to produce non-leaky and stable blood vessel formation mainly by the Tie2 receptor. Therefore, we envisioned that the application of cartilage oligomeric matrix protein-Ang1 (COMP-Ang1), a soluble, stable, and potent form of Ang1, would promote angiogenesis and provide a protective effect following unilateral middle cerebral artery occlusion (MCAO) in rats. To this end, we employed a 2-hour-MCAO model, and treated rats with adenovirus encoding COMP-Ang1 (Ade-COMP-Ang1) or control virus encoding beta-gal (Ade-beta-gal). Time course magnetic resonance images (MRIs) revealed significantly reduced infarct volume in the rats treated with Ade-COMP-Ang1 with an improvement of post-ischemic neurological deficits compared with rats treated with Ade-beta-gal. Moreover, compared to the rats treated with Ade-beta-gal, the rats treated with Ade-COMP-Ang1 showed an increase in blood vessels, especially in the border zone adjacent to the infarction, increased number of endogenous neuronal progenitor cells in the ischemic brain, and decreased number of TUNEL-positive cells. Taken together, COMP-Ang1 reduced infarct volume and consequently attenuated post-ischemic neurological deficits through enhanced angiogenesis and increased viable cell mass of neuronal cells.

PGF2α FP Receptor Contributes to Brain Damage Following Transient Focal Brain Ischemia

Although some of the COX-2 metabolites and prostaglandins have been implicated in stroke and excitotoxicity, the role of prostaglandin F(2alpha) (PGF(2alpha)) and its FP receptor have not been elucidated in the pathogenesis of ischemic-reperfusion (I/R) brain injury. Here we investigated the FP receptor's contribution in a unilateral middle cerebral artery (MCA) occlusion model of focal cerebral ischemia in mice. The MCA in wild type (WT) and FP knockout (FP(-/-)) C57BL/6 male mice was transiently occluded with a monofilament for 90 min. After 96 h of reperfusion, the FP(-/-) mice had 25.3% less neurological deficit (P < 0.05) and 34.4% smaller infarct volumes (P < 0.05) than those of the WT mice. In a separate cohort, physiological parameters were monitored before, during, and after ischemia, and the results revealed no differences between the groups. Because excitotoxicity is an acute mediator of stroke outcome, the effect of acute NMDA-induced neurotoxicity was also tested. Forty-eight hours after unilateral intrastriatal NMDA injection, excitotoxic brain damage was 20.8% less extensive in the FP(-/-) mice (P < 0.05) than in the WT counterparts, further supporting the toxic contribution of the FP receptor in I/R injury. Additionally, we investigated the effect of post-treatment with the FP agonist latanoprost in mice subjected to MCA occlusion; such treatment resulted in an increase in neurological deficit and infarct size in WT mice (P < 0.05), though no effects were observed in the latanoprost-treated FP(-/-) mice. Together, the results suggest that the PGF(2alpha) FP receptor significantly enhances cerebral ischemic and excitotoxic brain injury and that these results are of importance when planning for potential development of therapeutic drugs to treat stroke and its acute and/or long term consequences.

Recovery of spontaneous neuromagnetic activity after extracranial–intracranial bypass in a patient with middle cerebral artery occlusion

Cerebral blood flow and neuromagnetic activity were measured using (123)I-iodoamphetamine-single photon emission computed tomography (IMP-SPECT) and magnetoencephalography (MEG), before and after extracranial-intracranial (EC-IC) bypass surgery in a 55-year-old woman with unilateral middle cerebral artery occlusion that occurred with intraventricular haemorrhage. Frequency analysis of slow waves measured on MEG was performed using an adaptive beam-former method. Distribution of delta waves was observed pre-operatively corresponding to areas of cerebral hypo-perfusion as confirmed by IMP-SPECT but disappeared post-operatively with improvements in cerebral blood flow. Imaging of slow-wave distributions with MEG may represent a new technique for identifying cerebral ischaemia.

Chronic Middle Cerebral Artery Occlusion: A Hemodynamic and Metabolic Study with Positron-Emission Tomography

Chronic middle cerebral artery (MCA) occlusion is more common than generally thought. It is important to assess the cerebral hemodynamic status in patients with this chronic condition. We investigated the cerebral hemodynamic and metabolic disturbances in these patients in relation to the development of the collateral vasculature. We studied 13 patients with chronic unilateral MCA occlusion who had a minor or no stroke by using positron-emission tomography (PET). PET was performed by the oxygen 15 ((15)O) gas steady-state inhalation method. The intracranial arteries were evaluated by digital subtraction angiography. We divided the patients into 2 subgroups according to whether they had a normal or increased oxygen extraction fraction (OEF) in the occluded MCA territory and compared the 2 groups. Of the 13 patients, 9 were classified into the normal OEF and 4 were classified into the increased OEF group. In the increased OEF group, the mean OEF values were also increased in the territories of the ipsilateral anterior cerebral artery, ipsilateral posterior cerebral artery, and contralateral MCA. The patients in the increased OEF group had more than 1 steno-occlusive lesion in the major intracranial arteries (P = .008). Three of the 4 patients in the increased OEF group also had vascular lesions in the collateral pathways to the MCA territory. Most patients with chronic MCA occlusion did not show severe hemodynamic impairment. Those with increased OEF tended to have other areas of severe hemodynamic impairment and other vascular lesions, especially in the collateral pathways.

Ipsilateral and contralateral cortical apoptosis in rats after unilateral middle cerebral artery occlusion

Cerebral ischemia and subsequent reperfusion is known to induce irreversible tissue damage with the consequence of more or less pronounced impairments. The underlying cellular and molecular mechanisms however are far from being understood today. In the present study, we employed the well established rat model MCAO (middle cerebral artery occlusion), which uses a unilateral ligature of the middle cerebral artery for a defined period of time, prior to reperfusion. Animals develop symptoms, comparable to stroke in humans. Brains of such animals were used for histochemical and biochemical evaluation of markers, indicating programmed cell death (apoptosis) as evidenced by presence of tissue transglutaminase stained cells, DNA fragmentation and presence of cytochrome c in both cerebral hemispheres ipsilateral and contralateral to occlusion.

Acute Leakage Patterns of Fluorescent Plasma Flow Markers after Transient Focal Cerebral Ischemia Suggest Large Openings in Blood‐Brain Barrier

This study tested the hypothesis that blood-brain barrier (BBB) opening during acute reperfusion permits the passage of smaller macromolecules but not larger ones and that this molecular size restriction disappears over time. Following 3 hours (h) of unilateral middle cerebral artery occlusion and either 3 or 21 h of reperfusion, Wistar rats (n = 42) were injected with Evans blue (EB, a fluorescent dye that binds instantly to plasma albumin yielding EB-tagged albumin, EB-Alb) and with one of three fluorescent dextrans ranging in size from 77- to 2000-kDa. During occlusion and reperfusion, ischemic status of the affected tissue was confirmed by magnetic resonance imaging (MRI). Blood-to-brain transfer of the dextrans relative to that of EB-Alb was examined by fluorescence microscopy within three regions with ischemic damage. Increase in EB-Alb leakage from 3 to 21 h of reperfusion was significant (from 40-60% to 80-90% of fields examined; p < 0.05). Co-leakage of the largest dextran used 2000-kDa, with EB-Alb was observed in only 40% of the fields at 3+3 h, but nearly in all at 3 + 21 h (p < 0.01). Parenchymal distribution of the tracers differed among the fields and included considerable cellular uptake of EB-Alb and some of dextrans. Supporting the hypothesis, opening of the BBB was insufficient to allow passage of the largest dextran at 3 + 3 h in about 40% of the microvascular networks viewed. The number of total leaky microvascular beds increased by nearly 50% between 3 + 3 h and 3 + 21 h.

Endogenous neurogenesis and neovascularization in the neocortex of the rat after focal cerebral ischemia

The present study was designed to examine whether endogenous neurogenesis and neovascularization occur in the neocortex of the ischemic rat brain after unilateral middle cerebral artery occlusion (MCAO). Sprague-Dawley rats were divided into six groups (n = 29): one control group (n = 4) and five groups composed of animals sacrificed at increasing times post-MCAO (2 days and 1, 2, 4, and 8 weeks; n = 5 per group). To determine the presence of neurogenesis and neovascularization in the ischemic brain, nestin, Tuj1, NeuN, GFAP, Tie2, RECA, and 5-bromo-2'-deoxyuridine (BrdU) were analyzed immunohistochemically. In addition, nestin, GFAP, and Tie2 expression was determined by Western blotting. Triple-labeling of nestin, BrdU, and laminin was performed to visualize the interaction between endogenous neurogenesis and neovascularization. The number of BrdU- and nestin-colabeled cells increased markedly in the neocortex and border zone of the ischemic area up to 1 week after MCAO and decreased thereafter. Western blot analysis revealed that the expression of nestin, Tie-2, and GFAP was amplified in the ipsilateral hemisphere 2 days after MCAO and peaked 1 week after MCAO, compared with that in the normal brain. After ischemic injury, nestin- and BrdU-colabeled cells were observed in the vicinity of the endothelial cells lining cerebral vessels in the ipsilateral neocortex of the ischemic brain. Endogenous neurogenesis and neovascularization were substantially activated and occurred in close proximity to one other in the ipsilateral neocortex of the ischemic rat brain.

Dynamic susceptibility contrast perfusion magnetic resonance imaging in patients with symptomatic unilateral middle cerebral artery stenosis or occlusion

To evaluate cerebral hemodynamic disturbance in patients with symptomatic unilateral middle cerebral artery (MCA) high-grade stenosis or occlusion using dynamic susceptibility contrast perfusion magnetic resonance imaging (DSC-pMRI). DSC-pMRI was performed in 28 patients with symptomatic unilateral MCA high-grade stenosis or occlusion. Hemodynamic parameters including relative cerebral blood flow (rCBF), relative cerebral blood volume (rCBV), relative mean transit time (rMTT), and time to peak (TTP) were calculated and compared between the stenosed or occluded side and contralateral side. Seven of the 28 patients underwent surgery or interventional therapy, and hemodynamic changes between pre- and post-therapy were investigated. Prolonged rMTT and TTP were found in the stenosed or occluded MCA territories in all 28 patients. Significant differences were found in rCBV (P<0.05), rMTT (P<0.001), and TTP (P<0.001) between the stenosed or occluded side and the contralateral side. However, no significant difference was observed in rCBF (P>0.05). In the seven patients with surgical or interventional therapy, the values of rMTT and TTP significantly decreased after surgery or interventional therapy (P<0.001). DSC-pMRI may be a useful tool for evaluating and monitoring cerebral hemodynamic disturbance in patients with symptomatic unilateral MCA high-grade stenosis or occlusion.

Proton transfer ratio, lactate, and intracellular pH in acute cerebral ischemia

The amide proton transfer ratio (APTR) from the asymmetry of the Z-spectrum was determined in rat brain tissue during and after unilateral middle cerebral artery occlusion (MCAo). Cerebral lactate (Lac) as determined by (1)H NMR spectroscopy, water diffusion, and T(1rho) were quantified as well. Lac concentrations were used to estimate intracellular pH (pH(i)) in the brain during the MCA occlusion. A decrease in APTR during occlusion indicated acidification from 7.1 to 6.79 +/- 0.19 (a drop by 0.3 +/- 0.2 pH units), whereas pH(i) computed from Lac concentration was 6.3 +/- 0.2 (a drop by 0.8 +/- 0.2 pH units). Despite the disagreement between the two methods in terms of the size of the change in the absolute pH(i) during ischemia, DeltaAPTR and pH(i) (and Lac concentration) displayed a strong correlation during the MCAo. Diffusion and T(1rho) indicated cytotoxic edema following MCA occlusion; however, APTR returned slowly toward the values determined in the contralateral hemisphere post-ischemia. These data argue that the APTR during ischemia is affected not only by pH(i) but by other physicochemical factors as well, and indicates different aspects of pathology in the post-ischemic brain compared to those that influence water diffusion and T(1rho).