Magnetic resonance angiography of carotid and cerebral arterial occlusion in rats using a clinical scanner

In rat models to induce both focal cerebral ischemia and chronic cerebral hypoperfusion, it is highly desirable to verify the success of vessel occlusion and reopening with non-invasive method. The contrast-agent free 3D time-of-flight magnetic resonance angiography (TOF-MRA), diffusion-weighted imaging (DWI) and T2-weighted imaging by 3.0-T MR clinical scanner were applied when unilateral middle cerebral artery (MCA) was occluded and reopened, and after bilateral common carotid arteries were in ligation. The arterial angiograms of the rat brain and neck were achieved successfully in all chosen directions by the 3D TOF-MRA. It was shown that MCA in occlusion presented no signal in MRA, and the parenchyma of the ipsilateral MCA territory hypointensity signal in maps of apparent diffusion coefficient (ADC). After reperfusion, the signal intensity of ipsilateral MCA was resumed in MRA, and the decreased ADC was restored simultaneously. However, after 5h of reperfusion, it was found that the value of ADC deteriorated second time with high T2 value. In bilateral common carotid artery occlusion (BCCAO) rats, it can be confirmed by MRA that the effectively occluded BCCA presented the absent signal and the basilar artery became tortuous. As a result, MRA by clinical scanner was proved of a valuable method to validate transient middle cerebral artery occlusion (MCAO) and permanent BCCAO rat model.     

Chronic cerebral hypoperfusion induces vascular plasticity and hemodynamics but also neuronal degeneration and cognitive impairment

Chronic cerebral hypoperfusion (CCH) induces cognitive impairment, but the compensative mechanism of cerebral blood flow (CBF) is not fully understood. The present study mainly investigated dynamic changes in CBF, angiogenesis, and cellular pathology in the cortex, the striatum, and the cerebellum, and also studied cognitive impairment of rats induced by bilateral common carotid artery occlusion (BCCAO). Magnetic resonance imaging (MRI) techniques, immunochemistry, and Morris water maze were employed to the study. The CBF of the cortex, striatum, and cerebellum dramatically decreased after right common carotid artery occlusion (RCCAO), and remained lower level at 2 weeks after BCCAO. It returned to the sham level from 3 to 6 weeks companied by the dilation of vertebral arteries after BCCAO. The number of microvessels declined at 2, 3, and 4 weeks but increased at 6 weeks after BCCAO. Neuronal degeneration occurred in the cortex and striatum from 2 to 6 weeks, but the number of glial cells dramatically increased at 4 weeks after BCCAO. Cognitive impairment of ischemic rats was directly related to ischemic duration. Our results suggest that CCH induces a compensative mechanism attempting to maintain optimal CBF to the brain. However, this limited compensation cannot prevent neuronal loss and cognitive impairment after permanent ischemia.     

Chronic cerebral hypoperfusion protects against acute focal ischemia, improves motor function, and results in vascular remodeling

Atherosclerosis may cause severe stenosis of the arteries supplying the brain, which induces chronic cerebral hypoperfusion. Although an infarction often occurs in this area, it is uncertain how brain vessels respond to the chronic hypoperfusion or how the vascular responses are related to stroke severity when the area has been subjected to severe ischemia. To address these uncertainties, we induced chronic cerebral hypoperfusion in Sprague-Dawley rats with a bilateral common carotid artery ligation (BCAL). A middle cerebral artery occlusion/reperfusion (MCAO/R) was introduced with a nylon suture four weeks after either BCAL (BCAL-MCAO) or a sham operation (Sham-MCAO). Motor disability scores and infarct sizes, based on 2,3,5-triphenyltetrazolium chloride staining, were significantly reduced with BCAL-MCAO treatment compared with sham-MCAO treatment (P<0.01). The diameters of the posterior cerebral, posterior communicating, and basilar arteries on the brain surface were larger and more tortuous in BCAL-treated rats (P<0.01). The density of large capillary- and arteriole-sized vessels in the brain parenchyma also increased in BCAL-treated rats (P<0.05). Strokes were less severe when the vicinity subjected to infarction was preconditioned with chronic cerebral hypoperfusion. Increasing the vascular reserve with adaptive vascular remodeling may have contributed to this response.     

Moderate loss of cerebellar Purkinje cells after chronic bilateral common carotid artery occlusion in rats

Pathological effects of moderate ischemia (oligemia, hypoperfusion) are relevant in relation to vascular factors in dementia. Chronic bilateral common carotid artery occlusion (BCCAO) in adult Wistar rats induces oligemia and leads to acute changes in gene expression, subacute changes in cortical astrocytes and prolonged changes in white matter tracts, while largely sparing neurons in the forebrain areas. Dilation and remodeling of the basilar artery ensures blood flow to the forebrain. The present study examined the hypoxia-sensitive Purkinje cells in the cerebellum after 6 months of BCCAO using conventional neuropathological analysis, immunohistochemistry and high-precision design-based stereologic methods. Purkinje cells in the vermis region revealed abnormally shaped nuclei. A stereologic analysis showed that the mean total number of Purkinje cells within the vermis was statistically significantly smaller in the BCCAO animals than in the control animals (d = 11.8%; P < 0.0001). BCCAO had no significant effect on the mean volumes of the molecular layer, granule cell layer and white matter in the vermis or the entire cerebellum. Remodeling of the basilar artery indicated that secondary vascular perturbations might be responsible for the effects of BCCAO on the cerebellar Purkinje cells.     

Minocycline attenuates white matter damage in a rat model of chronic cerebral hypoperfusion

White matter lesions are thought to result from chronic cerebral ischemia and constitute a core pathology of subcortical vascular dementia. This rarefaction has been known to be associated with microglial activation. We investigated whether minocycline, a microglial inhibitor, attenuates the white matter damage induced by chronic cerebral hypoperfusion that is used as a model of vascular dementia. Male Wistar rats were subjected to bilateral, permanent occlusion of the common carotid arteries (BCCAO) to induce chronic cerebral hypoperfusion. Minocycline or saline was injected daily for 2 weeks after BCCAO. In the corpus callosum and the optic tract, white matter damage observed with Klüver-Barrera staining was significantly attenuated in the minocycline-treated group compared to saline-treated controls. In control rats, immunoreactivities of major basic protein (MBP), Ox-42 as a microglial marker, and matrix metalloproteinase (MMP)-2 were increased in the corpus callosum. Minocycline significantly reduced these changes. Co-expression of Ox-42 and MMP-2 was confirmed by double immunofluorescence histochemistry. Our results suggest that chronic treatment with minocycline could be protective against at least some ischemic white matter damage, and its mechanism may be related to suppressing microglial activation.     

The plasticity of posterior communicating artery influences on the outcome of white matter injury induced by chronic cerebral hypoperfusion in rats

Abstract

Objective: This study was performed to identify which factors are involved in the development of white matter lesions induced by chronic cerebral hypoperfusion in rats.

Methods: Male Wistar and Sprague–Dawley (SD) rats (250–270 g) were subjected to the permanent occlusion of bilateral common carotid arteries (BCCAO). The regional cerebral blood flow (rCBF) was measured by laser Doppler flowmetry and the plasticity of the posterior communicating artery (PcomA) was visualized by transcardiac perfusion of latex solution. For the histological examination, Klüver–Barrera staining was used to evaluate white matter damage.

Results: When compared with SD rats, Wistar rats showed lower rCBF after BCCAO, as well as thinner PcomAs. Moreover, 21 days after BCCAO, Wistar rats showed marked vacuolation of white matter in the optic tract, whereas SD rats had an almost intact optic tract.

Discussion: These results suggest that the plasticity of the PcomA and the reduction of rCBF in Wistar rats are important factors in the development of BCCAO-induced white matter lesions.     

Hemodynamic and metabolic effects of extracranial carotid disease

Cerebral blood flow (CBF), cerebral blood volume (CBV), the CBF/CBV ratio - an index of the hemodynamic reserve capacity - the rate of oxygen metabolism (CMRO2), and the fractional extraction of oxygen by the brain (OEF) were studied by positron emission tomography (PET) in the cortical territory of both internal carotid arteries in 15 cases of transiently symptomatic or progressive extracranial atherosclerotic carotid disease. None of the patients had a major stroke or had a significant neurological deficit except 1 whose damaged hemisphere is excluded from study. All were asymptomatic at the time of PET scanning. Values were obtained in the middle cerebral artery (MCA) distribution, and in the anterior and posterior borderzone regions. Eight cases had unilateral carotid stenosis of 80% or greater and 7 had unilateral or bilateral occlusion of the origin of the internal carotid artery. Results obtained in patients were compared using Student's t-test, to those obtained in neurologically normal, elderly volunteers. Patients with carotid stenosis had a significantly decreased CBF (p less than .025) and CBF/CBV ratio (p less than .025) selectively in the anterior borderzone regions. This was accompanied by a trend toward elevated OEF and declining CMRO2 values. Patients with carotid occlusion had significantly decreased CBF (p less than .005), decreased CBF/CBV ratio (p less than .005) and decreased CMRO2 (p less than .025) in the ipsilateral anterior borderzone and MCA territories. Similar changes were present in the opposite hemisphere of patients with bilateral carotid disease. These results indicate that carotid stenosis is associated with hypoperfusion and diminished hemodynamic reserve capacity in the anterior borderzone, and that carotid occlusion produces more widespread hypoperfusion and metabolic depression.     

Pure sensory stroke revealing a complex malformation of extra- and intracranial cerebral arteries

Pure sensory strokes (PSS) are usually due to thalamic lacunar infarcts. We report a case of PSS in a 34-year-old female suffering from migraine who complained of sudden isolated left-sided paresthesia involving face, upper and lower limbs. We found a complex vascular abnormality associating internal carotid artery occlusion, posterior cerebral artery agenesia, aneurysm at the posterior face of the carotid siphon and angiomatous rete at the top of the basilar artery. The acquired origin of the carotid artery occlusion by possible neonatal dissection is probable since this patient has a permeable carotid canal on CT scan.     

Astrocytes react to oligemia in the forebrain induced by chronic bilateral common carotid artery occlusion in rats

The effects of oligemia (moderate ischemia) on the brain need to be explored because of the potential role of subtle microvascular changes in vascular cognitive impairment and dementia. Chronic bilateral common carotid artery occlusion (BCCAO) in adult rats has been used to study effects of oligemia (hypoperfusion) using neuropathological and neurochemical analysis as well as behavioral tests. In this study, BCCAO was induced for 1 week, or 2, 4, and 6 months. Sensitive immunohistochemistry with marker proteins was used to study reactions of astrocytes (GFAP, nestin), and lectin binding to study microglial cells during BCCAO. Overt neuronal loss was visualized with NeuN antibodies. Astrocytes reacted to changes in the optic tract at all time points, and strong glial reactions also occurred in the target areas of retinal fibers, indicating damage to the retina and optic nerve. Astrocytes indicated a change in the corpus callosum from early to late time points. Diffuse increases in GFAP labeling occurred in parts of the neocortex after 1 week of BCCAO, in the absence of focal changes of neuronal marker proteins. No significant differences emerged in the cortex at longer time points. Nestin labeling was elevated in the optic tract. Reactions of microglia cells were seen in the cortex after 1 week. Measurements of the basilar artery indicated a considerable hypertrophy, indicative of macrovascular compensation in the chronic occlusion model. These results indicate that chronic BCCAO and, by inference, oligemia have a transient effect on the neocortex and a long-lasting effect on white matter structures.     

A new model of white matter injury in neonatal rats with bilateral carotid artery occlusion

Periventricular leukomalacia is an important cause of cerebral palsy and characterized by cysts and coagulation necrosis in the periventricular white matter. Since no model of periventricular leukomalacia has been established in small animals, it is expected to establish a new model of white matter injury in immature rodents. Bilateral carotid arteries were occluded in neonatal rats at 5 days of age, and the brain neuropathologically examined at 7 days of age. Among 22 brains histologically examined, 20 (90.9%) had white matter changes including coagulation necrosis and cystic lesions in and around the internal capsule, while only two had small cerebral infarction and five showed some ischemic neurons in the cerebral cortex. Cerebral blood flow (CBF) decreased to about 25% of controls in the subcortical white matter in the animals with bilateral carotid artery occlusion (BCAO). Amyloid precursor protein (APP) immunohistochemistry demonstrated various APP-immunoreactive axonal profiles in the internal capsule and the subcortical white matter, and stronger expression of APP in pyramidal neurons in the cerebral cortex of BCAO brains. These results indicated that the white matter is more vulnerable than the cerebral cortex in 5-day-old rats when CBF decreases to about 25% and suggested that this model is useful for investigating the white matter changes induced by cerebral hypoperfusion in the neonatal brain, since previous models of hypoxic-ischemic brain injury in neonatal mice and rats revealed preferential susceptibility of the gray matter. It was also indicated that APP is a sensitive marker for mild axonal disruption in the white matter of the immature brain.     

Multiple aneurysms associated with bilateral carotid occlusion and venous angioma: surgical management risk-case report.

A unique case of multiple aneurysms associated with bilateral carotid artery occlusion and venous angioma is described. A 42 year old female presented with subarachnoid haemorrhage. Cerebral angiograms demonstrated(1) a ruptured saccular aneurysm in the right posterior cerebral artery,(2) bilateral occlusion of internal carotid arteries,(3) a rete mirabile in the subtemporal fossa fed by left external carotid artery which connected with the internal carotid artery at the cavernous portion where a saccular aneurysm had formed, and(4) a venous angioma in the posterior fossa. The ruptured aneurysm of the posterior cerebral artery was obliterated preserving the anterior choroidal arteries. However, a left hemiparesis developed and CT scan revealed a small low density area in the right posterior limb of the internal capsule postoperatively. A ruptured aneurysm associated with bilateral extracranial carotid occlusion poses a clinical dilemma and treatment of such cases is challenging and difficult. The non-surgical and surgical outcomes of ruptured cerebral aneurysms associated with internal carotid occlusion are reviewed.     

Decreased cerebral perfusion and oxidative stress result in acute and delayed cognitive impairment

Chronic cerebral hypoperfusion (CCH) is common in the pathogenesis of cognitive impairment, in which oxidative stress plays an important role. Here we describe an alternative rat model for CCH that involves two-stage, three-vessel occlusion (2s-3VO) and compare its effects with those of permanent bilateral occlusion (2VO) of the common carotid arteries. Real-time cerebral blood flow (CBF) during the surgery was monitored. Spatial learning and memory were tested with the Morris water maze, and oxidative damage was evaluated by measuring malondialdehyde (MDA) levels in both the hippocampus and cortex. We found that the CBF drop in the early stage of the 2s-3VO model was more modest than that in the 2VO model. Like 2VO rats, 2s-3VO rats showed impaired spatial learning and memory and increased MDA levels 8 weeks after surgery. Interestingly, when pooling observations from previous studies, we confirmed that oxidative damage appeared later than spatial learning and memory deficits but lasted longer than did cerebral hypoperfusion. Thus, the 2s-3VO model appears to be a suitable model for the study of CCH. Moreover, data support the notion that cognitive impairment in CCH rat models may be induced early by cerebral hypoperfusion early and in a later phase by oxidative stress.     

Early operation of ruptured basilar artery aneurysm associated with bilateral carotid occlusion (moyamoya disease)

A 34-year-old Caucasian man presented with subarachnoid hemorrhage. Angiography revealed bilateral carotid occlusion at the cavernous sinus and an aneurysm at the basilar artery bifurcation. The whole brain was supplied with blood from the basilar artery and posterior cerebral arteries through a large number of collateral vessels to the internal carotid artery bifurcation, middle cerebral and anterior cerebral arteries: the moyamoya phenomenon. The aneurysm was clipped within hours of the subarachnoid hemorrhage. The relation between moyamoya disease and basilar artery aneurysms is discussed and some surgical and management considerations are given.     

Magnetic resonance angiography of the extracranial carotid arteries and intracranial vessels: a review

MRI is uniquely suited for evaluation of vascular structures due to its sensitivity to a variety of flow-related phenomena. Recent work has demonstrated that high quality magnetic resonance angiograms (MRA) of the carotid arteries and intracranial vasculature can be achieved by using gradient-echo techniques with short echo times. These MRAs are displayed like conventional arteriograms, but are acquired in a noninvasive fashion with a minimal increase in examination time. We used MRA to visualize 50 of 54 carotid bifurcations tested, with good correlation to the intra-arterial angiograms. We examined the intracranial vasculature in over 40 patients, and demonstrated aneurysms, vascular malformations, and occlusions.     

Communications between vertebro-basilar and carotid arterial circulations in the gerbil

The cerebrovascular anatomy of the gerbil was evaluated to assess the existence and significance of arterial communications at the base of the brain. Postmortem intra-aortic dye injection in ten gerbils demonstrated communications between both anterior cerebral arteries and between the basilar and posterior cerebral arteries. Vessels of the carotid circulation filled with dye even when both common carotid arteries were occluded. Intravenous injection of carbon black in vivo produced cerebral staining in six of ten gerbils that were previously subjected to bilateral common carotid artery occlusion, thereby demonstrating that such occlusion does not invariably produce complete cerebral ischemia. Variability in ischemia, possibly related to differences in the number and size of these communicating vessels, may influence the outcome and interpretation of experiments utilizing carotid occlusion in the gerbil.     

慢性脑低灌注大鼠海马缝隙连接蛋白32和36的表达变化

目的：观察慢性脑低灌注大鼠海马组织中缝隙连接蛋白（Cx）32和Cx36的变化,探讨其在慢性脑低灌注认知功能损害中的作用。方法：双侧颈总动脉永久性结扎（2VO）制作慢性脑低灌注模型,Morris水迷宫检测2VO大鼠空间学习记忆能力变化,免疫组化检测2VO大鼠海马各区Cx32和Cx36的表达水平。结果：慢性脑低灌注大鼠的空间学习记忆能力较假手术对照组显著下降,海马Cx32和Cx36的表达也有降低。结论：Cx32和Cx36可能参与了慢性脑低灌注导致的认知功能损害。     

Arteriogenesis in hypoperfused rat brain.

Experimental and clinical studies have provided evidence for spontaneous and therapeutically induced arteriogenesis after occlusion of major peripheral or cardiac vessels. Such evidence is lacking for the cerebrovascular system. In halothane-anesthetized rats, different degrees of brain hypoperfusion were induced by one- to four-vessel occlusion, that is, one or both common carotid arteries in combination with or without bilateral vertebral artery occlusion. The flow decline was monitored by laser Doppler flowmetry, the residual hemodynamic reserve by testing flow reactivity to ventilation with 6% CO(2) and arteriogenesis by intravascular latex infusion and immunohistochemistry of vascular proliferation and monocyte adhesion. The optimum condition for induction of arteriogenesis was three-vessel (one carotid plus both vertebral arteries) occlusion, which led to reduction of blood flow to about 50% and complete suppression of CO(2) reactivity, but no histologic injury. One week after three-vessel occlusion, the ipsilateral posterior cerebral artery significantly enlarged by 39%, and after 3 weeks by 72%, paralleled by the partial return of CO(2) reactivity and the appearance of immunohistochemical markers of arteriogenesis. Three-vessel occlusion is a reliable model for the induction of arteriogenesis in the adult brain and is a new approach for exploring the potentials of arteriogenesis for the prevention of progressing brain ischemia.     

Hypertension impairs leptomeningeal collateral growth after common carotid artery occlusion: restoration by antihypertensive treatment

Chronic mild hypoperfusion has been shown to enlarge pial collateral vessels in normal mouse brains. The purpose of this study was to clarify the effect of hypertension on pial collateral vessel development after chronic hypoperfusion using spontaneously hypertensive rats (SHR). In normotensive rats, unilateral common carotid artery (CCA) occlusion enlarged leptomeningeal collateral vessels. CCA occlusion also preserved residual cerebral blood flow (CBF) and attenuated infarct size after middle cerebral artery (MCA) occlusion 14 days later. In contrast, in SHR, CCA occlusion neither enlarged the leptomeningeal anastomosis nor showed protective effects after MCA occlusion. However, decreasing blood pressure using an angiotensin II AT1 receptor blocker restored the beneficial effect of CCA occlusion on collateral growth as well as on residual CBF and infarct size after MCA occlusion. Adaptive responses in CBF autoregulation curves observed 14 days after CCA occlusion in normotensive rats were impaired in untreated SHR, but were restored after antihypertensive treatment. In conclusion, SHR have impaired leptomeningeal collateral growth after CCA occlusion, but antihypertensive treatment restores the beneficial effect of CCA occlusion on collateral circulation.     

Impact of intracranial blood-flow redistribution on stroke size during ischemia-reperfusion in 7-day-old rats

We evaluated color-coded pulsed Doppler ultrasound imaging for the assessment of intracranial blood flow in two models of cerebral ischemia in 7-day-old (P7) rats. Blood-flow velocities (BFVs) were measured in the internal carotid arteries and basilar trunk upstream from the circle of Willis, and in the posterior cerebral arteries downstream (1) before, (2) during left middle cerebral artery electrocoagulation and 50 min-transient either one (I/R-1) or both (I/R-2) common carotid (CCA) arteries occlusion, and (3) after release of CCA(s) occlusion. At 48 h after ischemia 41-48% (I/R-1 model) and 24% (I/R-2 model) of rats did not present a lesion. Those rats displayed increased mean BFV in both right internal carotid artery and basilar trunk in I/R-1 model, and increased mean BFV in the basilar trunk (BT) in I/R-2 model. In contrast, no significant changes in mean BFV were observed in lesioned rats. Furthermore, mean BFV in the BT was inversely correlated to the size of the lesion (r2 = 0.75, p<0.0001) in the I/R-2 model. Thus, we demonstrated the protective role of collateral support in P7 rodents. Ultrasound imaging can evidence the establishment or not of the cerebral collateral recruitment, leading to the presence or absence of a lesion. This novel approach should greatly help preclinical studies to reduce animal variability.     

慢性前脑缺血致血管性痴呆大鼠不同脑区MAP-2的经时变化研究

目的研究微管结合蛋白-2(MAP-2)在慢性前脑缺血致血管性痴呆大鼠额、颞叶皮质和海马区的经时变化。方法采用双侧颈总动脉永久结扎方法制备慢性前脑缺血致VD大鼠动物模型;采用免疫组织化学方法检测痴呆鼠不同脑区MAP-2的经时变化情况;采用同济大学研制的HPIAS-1000高清晰彩色病理图文分析系统对不同脑区MAP-2阳性信号的面积密度进行分析。结果实验发现痴呆大鼠不同脑区MAP-2的表达随时间变化。缺血1个月、2个月、4个月后MAP-2的表达阳性面积逐渐减少,与对照组相比有统计学意义。从形态学变化角度来说,1个月时MAP-2轻度变化,提示为可逆性损伤。随着缺血时间的延长,MAP-2免疫组化染色越来越淡。至4个月时,MAP-2免疫组化染色基本消失,提示为不可逆损伤。结论慢性持续性脑血流量下降致MAP-2的改变在VD的病理生理过程中起重要作用。