No rarefaction of cerebral arterioles in hypertensive rats

A reduction in the density of small arterioles (rarefaction) has been reported in several vascular beds of the spontaneously hypertensive rat (SHR). There have been conflicting reports on the existence of rarefaction in the pial vasculature of SHR. In this study, we determined whether there was rarefaction of pial arterioles in several models of hypertension. We studied SHR; two-kidney, one-clip Goldblatt hypertensive rats; deoxycorticosterone-salt hypertensive rats; and Dahl salt-sensitive rats fed high salt diet. The two groups of normotensive controls were Wistar--Kyoto rats and Dahl salt-sensitive rats fed low salt diet. The duration of hypertension was about 2 months. Density of first-, second-, third-, and fourth-order arterioles was determined by counting the number of vessels from enlarge photographs. We also measured the lengths of segments of the arterioles. We did not observe any evidence of rarefaction of arterioles in the pial vasculature in any of the hypertensive groups of rats. We conclude that (i) rarefaction of arterioles does not occur in the pial microvasculature after approximately 2 months of hypertension and (ii) rarefaction of pial arterioles does not account for abnormalities in the cerebral circulation of hypertensive rats such as protection of the blood-brain barrier or changes in autoregulation of cerebral blood flow.     

Indian-Ink Perfusion Based Method for Reconstructing Continuous Vascular Networks in Whole Mouse Brain

The topology of the cerebral vasculature, which is the energy transport corridor of the brain, can be used to study cerebral circulatory pathways. Limited by the restrictions of the vascular markers and imaging methods, studies on cerebral vascular structure now mainly focus on either observation of the macro vessels in a whole brain or imaging of the micro vessels in a small region. Simultaneous vascular studies of arteries, veins and capillaries have not been achieved in the whole brain of mammals. Here, we have combined the improved gelatin-Indian ink vessel perfusion process with Micro-Optical Sectioning Tomography for imaging the vessel network of an entire mouse brain. With 17 days of work, an integral dataset for the entire cerebral vessels was acquired. The voxel resolution is 0.35×0.4×2.0 µm³ for the whole brain. Besides the observations of fine and complex vascular networks in the reconstructed slices and entire brain views, a representative continuous vascular tracking has been demonstrated in the deep thalamus. This study provided an effective method for studying the entire macro and micro vascular networks of mouse brain simultaneously.     

Aquaporin-1 in blood vessels of rat circumventricular organs

Although the water channel protein aquaporin-1 (AQP1) is widely observed outside the rat brain in continuous, but not fenestrated, vascular endothelia, it has not previously been observed in any endothelia within the normal rat brain and only to a limited extent in the human brain. In this immunohistochemical study of rat brain, AQP1 has also been found in microvessel endothelia, probably of the fenestrated type, in all circumventricular organs (except the subcommissural organ and the vascular organ of the lamina terminalis): in the median eminence, pineal, subfornical organ, area postrema and choroid plexus. The majority of microvessels in the median eminence, pineal and choroid plexus, known to be exclusively fenestrated, are shown to be AQP1-immunoreactive. In the subfornical organ and area postrema in which many, but not all, microvessels are fenestrated, not all microvessels are AQP1-immunoreactive. In the AQP1-immunoreactive microvessels, the AQP1 probably facilitates water movement between blood and interstitium as one component of the normal fluxes that occur in these specialised sensory and secretory areas. AQP1-immunoreactive endothelia have also been seen in a small population of blood vessels in the cerebral parenchyma outside the circumventricular organs, similar to other observations in human brain. The proposed development of AQP1 modulators to treat various brain pathologies in which AQP1 plays a deleterious role will necessitate further work to determine the effect of such modulators on the normal function of the circumventricular organs.     

The vascular morphometry of some brain stem parts

With the view of following the vascular system of the cerebral trunk, there have been simultaneously injected coloured tracers combined with radio-opaque substances and radio-isotopes (32P, 22Ca) by intravital perfusion to 130 animals (dogs, rabbits and rats). The macro- and microscopical vasculature of the human brain stem was studied on 30 specimens of adults and children. The angioarchitecture of the diverse parts and sectors of the brain stem was studied as a whole in tetraline transparentised preparations and on histological section in comparison with the microradiographies and to incorporation of radioisotopes. A particular attention has been given to the irrigation of hypoglossal nuclei, the bulbar oliva, and the pontopeduncular region, all being zones with multiple sources of vascularization. The vascular morphometry has been macro- and microscopically performed, the data being worked out mathematically-statistically by computing some hemodynamical indices in order to estimate the circulation in different zones of the brain stem.     

Quantitative assessments of cerebral vascular damage with a silicon rubber casting method in photochemically-induced thrombotic stroke rat models

Previous studies have described microvascular disturbances downstream of occluded large vessels arising during the acute phase (several hours) following cerebral ischemic insult. Prolonged microvascular disturbances may cause delayed neuronal cell death in ischemic penumbral regions, leading to expanded brain infarctions and poor neurological and functional outcomes. The lack of simple and quantitative methods for investigating this microcirculation failure suggests the need to develop a new method for clarifying the precise distribution and persistence of post-ischemic microvascular disturbances. The present study used a silicone rubber casting method in quantitative analyses of microvascular conditions in photochemically-induced thromboembolic (PIT) stroke rat models. After the casting procedure in rats with PIT stroke, a 6 microm-thick coronal section was obtained, and quantitative analyses of microvascular density and measurements of the infarct area in the serial section were performed. The major findings of the present study are as follows: (1) Silicone rubber casting techniques can be applied to precise quantitative analyses of microvessels in the same individual in whom brain infarct volume was measured; (2) the persistence and spatial distribution of microvascular disturbances assessed at the ischemic core, ischemic penumbra, and non-ischemic regions strongly suggest that microvascular disturbances affect brain infarct expansion; (3) the current method demonstrated the protective effects of MK-801 on microvessels, indicating that the technique may be useful in investigating factors that provide vascular protection. The experimental procedure introduced here would facilitate future evaluations of vascular protective agents.     

Mathematical simulation of gliding contusions

A gliding contusion in the acute phase is characterized by a streaklike hemorrhage of venous origin situated subcortically in a paramedial convolution. In later stages perivascular necrosis may develop. This type of injury is caused by head angular acceleration and is often seen in traffic casualties when the head has hit the steering wheel, the dashboard or the windshield.

The deformation of the brain matter close to the superior sagittal sinus has been simulated by means of a mathematical viscoelastic model in order to clarify the genesis of the gliding contusions. The blood vessels in the brain matter will be strained as a consequence of the brain deformation which results from head angular acceleration. The highest values of the strain occur subcortically where the blood vessels are injured first. The tolerance levels for gliding contusions have been determined. The calculations which were based on experiments regarding the dynamic properties of the superior cerebral veins and on two alternative injury criteria proposed, indicate that a gliding contusion is not likely to arise if the maximal angular acceleration does not exceed 4500 rad/sec² or the change in angular velocity does not exceed 70 rad/sec.     

Vascular damage after fractionated whole-brain irradiation in rats

Whole-brain irradiation of animals and humans has been reported to lead to late delayed structural (vascular damage, demyelination, white matter necrosis) and functional (cognitive impairment) alterations. However, most of the experimental data on late delayed radiation-induced brain injury have been generated with large single doses or short fractionation schemes that may provide a less accurate indication of the events that occur after clinical whole-brain radiotherapy. The pilot study reported here investigates cerebral vascular pathology in male Fischer 344 rats after whole-brain irradiation with a fractionated total dose of 137Cs gamma rays that is expected to be biologically similar to that given to brain tumor patients. The brains of young adult rats (4 months old) were irradiated with a total dose of 40 Gy, given as eight 5-Gy fractions twice per week for 4 weeks. Brain capillary and arteriole pathology was studied using an alkaline phosphatase enzyme histochemistry method; vessel density and length were quantified using a stereology method with computerized image processing and analysis. Vessel density and length were unchanged 24 h after the last dose, but at 10 weeks postirradiation, both were substantially decreased. After 20 weeks, the rate of decline in the vessel density and length in irradiated rats was similar to that in unirradiated age-matched controls. No gross gliosis or demyelination was observed 12 months postirradiation using conventional histopathology techniques. We suggest that the early (10-week) and persistent vascular damage that occurs after a prolonged whole-brain irradiation fractionation scheme may play an important role in the development of late delayed radiation-induced brain injury.     

Age and changes in human brain capillaries (histochemical study)

The vascular-capillary bed in the field 17 of the human brain cortex (from fetuses up to 86 years of age) has been studied by means of some histochemical methods to reveal alcaline phosphatase and Mg-ATPase. As demonstrate quantitative methods, active processes of the capillary bed formation in the cerebral cortex take place before 8-16 years of age. Towards the old age, the amount of the vessels with a high and moderate enzymatic activity decreases, but those with a low enzymatic activity increases. The average parameters of the optic density of the reaction product decreases respectively in the capillary wall, so does the intensity of metabolic processes, while the value of the working surface increases up to 75 years of age.     

Neuronal expression of the Ccm2 gene in a new mouse model of cerebral cavernous malformations

Cerebral cavernous malformations are vascular defects of the central nervous system consisting of clusters of dilated vessels that are subject to frequent hemorrhaging. The genes mutated in three forms of autosomal dominant cerebral cavernous malformations have been cloned, but it remains unclear which cell type is ultimately responsible for the lesion. In this article we describe mice with a gene trap insertion in the Ccm2 gene. Consistent with the human phenotype, heterozygous animals develop cerebral vascular malformations, although penetrance is low. β-galactosidase activity in heterozygous brain and in situ hybridization in wild-type brain revealed Ccm2 expression in neurons and choroid plexus but not in vascular endothelium of small vessels in the brain. The expression pattern of Ccm2 is similar to that of the Ccm1 gene and its interacting protein ICAP1 (Itgb1bp1). These data suggest that cerebral cavernous malformations arise as a result of defects in the neural parenchyma surrounding the vascular endothelial cells in the brain. Nicholas W. Plummer, Teresa L. Squire and Sudha Srinivasan contributed equally to this work.     

一氧化氮对脑血流的调节

一氧化氮是近年来发现的一种重要的血管活性因子,它通过激活平滑肌细胞内水溶性鸟苷酸环化酶,而产生血管舒张作用,在正常生理条件下,ＮＯ不仅对外因管有作用,对脑血管也有作用,但关于它在低氧和高二氧化碳条件下脑血管是否具有调节作用还存在着争议。     

Microcirculatory bed of the bulbar conjunctiva in 3- to 9-month-old fetuses

In 32 corpses of 3-9-month-old fetuses the diameter, length amount of the microcirculatory bed vessels and their wall thickness have been studied in different areas of the eye bulb conjunctiva by means of impregnation after Kuprianov. General morphofunctional vascular peculiarities have been revealed. They are connected with age and structure of the developing tissues in the eye bulb conjunctiva. Two stages of development have been defined: 3-6 and 7-9 months.     

Testosterone 5 alpha-reductase in discrete hypothalamic nuclear areas in the rat: effect of castration

The conversion of testosterone into 5 alpha-dihydrotestosterone (DHT) has been studied in different hypothalamic nuclear areas and in the superficial layers of the cerebral cortex of normal and castrated male rats. The tissue fragments utilized in each incubation have been punched from frozen brain sections utilizing calibrated needles. Castration has been performed 12 (short term) and 180 (long term) days before sacrifice. The nuclear areas studied include: the medial preoptic nucleus (MPN), the lateral preoptic nucleus (LPN), the anterior hypothalamic nucleus (AHN), the lateral hypothalamic nucleus (LHN), the posterior hypothalamic nucleus (PHN), the nucleus ventromedialis (HVM), the arcuate nucleus (AR), the median eminence (ME), the nucleus paraventricularis (HPV), the supraoptic nucleus (SO) and the suprachiasmatic nucleus (SC). The possible effect of castration on the 5 alpha-reductase, were assessed in the MPN,LPN,AHN,LHN,PHN and in the cerebral cortex. The results indicate that, in the male rat: 1) the lateral preoptic(LPN) and the lateral hypothalamic nuclei(LHN) possess a 5 alpha-reductase activity higher than that present in the cerebral cortex and in the other hypothalamic nuclei considered; 2)the suprachiasmatic nucleus (SC) apparently possesses a testosterone metabolizing activity lower than that found in any other nervous structures studied so far; 3) castration does not seem to influence the 5 alpha-reductase activity either in the hypothalamic nuclear structures considered or in the cerebral cortex.     

The effect of the calcium antagonist nimodipine upon local cerebral glucose utilization in the rat brain

The new calcium antagonist Nimodipine has been shown to have more powerful dilator action on cerebral than peripheral vessels. The effect of the drug on cerebral metabolism was studied in conscious rats using the /14C/-2-deoxyglucose quantitative autoradiographic technique. Intravenous injection of Nimodipine, 2 mcg/Kg, determined significant increases in local cerebral glucose utilization that appeared to be homogeneous in magnitude and anatomic distribution throughout the brain. This study raises the question whether Nimodipine affects brain functions by other mechanisms than an increase in cerebral blood flow.     

Comparative study of the cholinergic and adrenergic innervation of the cerebral vessels in humans and various laboratory animals

Choline- and adrenergic innervation of intracerebral branches of the middle and posterior cerebral arteries has been investigated histochemically and electron microscopically in slices, or after their preparation after W. Penfield. The vessels have been studied in the area of the fields 41, 17 of the cerebrum, trunk and spinal cord of the human being, cat and dog. When studying innervation of the intracerebral arteries (ICA), the preparation method has some advantages in comparison with investigation of these vessels in slices of the brain. Around most of the ICA from 200 up to 30 mcm in the diameter choline- and adrenergic nervous conductors are revealed. Using the method for calculating varicosities in nervous plexuses, it is demonstrated that degree of the ICA innervation is two times less than that in the arteries of similar caliber in the cerebral pia mater.     

Effect of aging on neurotransmitter receptor binding in rat and human brain.

β-Adrenergic and GABA receptor binding were measured in brain areas of rats 3 to 24 months of age. While GABA receptor binding was not significantly different across age in any area, β-adrenergic receptor binding was significantly reduced in the cerebellum and brain stem, but not cerebral cortex, of 24-month-old animals. The loss in β-adrenergic receptor binding does not correlate in a temporal fashion with the reported decrease in norepinephrine-stimulated cyclic AMP accumulation in the cerebellum which occurs as early as 12 months of age. An age-related reduction in β-adrenergic binding was also noted in human cerebellar tissue obtained at autopsy, suggesting that the cerebellar dysfunction seen with aging may be related to a loss of cerebellar neurons which receive noradrenergic input.     

Lateralized behaviour of a non-social cichlid fish (Amatitlania nigrofasciata) in a social and a non-social environment

Cerebral lateralization, the partitioning of cognitive function preferentially into one hemisphere of the brain, is a trait ubiquitous among vertebrates. Some species exhibit population level lateralization, where the pattern of cerebral lateralization is the same for most members of that species; however, other species show only individual level lateralization, where each member of the species has a unique pattern of lateralized brain function. The pattern of cerebral lateralization within a population and an individual has been shown to differ based on the stimulus being processed. It has been hypothesized that sociality within a species, such as shoaling behaviour in fish, may have led to the development and persistence of population level lateralization. Here we assessed cerebral lateralization in convict cichlids (Amatitlania nigrofasciata), a species that does not shoal as adults but that shoals briefly as juveniles. We show that both male and female convict cichlids display population level lateralization when in a solitary environment but only females show population level lateralization when in a perceived social environment. We also show that the pattern of lateralization differs between these two tasks and that strength of lateralization in one task is not predictive of strength of lateralization in the other task.     

Changes in dehydrogenase glutamate in the rat brain caused by thyroid hormone deficiency

The dependent GDH-NADPH activity in adenohypophysis and other cerebral areas, has been studied in hypothyroid rats, in which hypothyroidism has been induced surgically. After thyroidectomy a decrease of GDH activity in limbic system (amygdala, septum and hippocampus), and an increase of this enzyme in cortex and hypothalamus have been found, with no changes in adenohypophysis. The alterations of GDH activity, induced by thyroidectomy, have been corrected, although not uniformly in the different brain areas after L-T3 treatment.     

Vascular growth factors in cerebral ischemia

During the past decade, there has been a surge of interest in growth factors (GFs) that act selectively on vascular endothelium and perivascular cells. Studies employing mutant mice or the administration of recombinant proteins have suggested that these factors not only mediate the proliferation of endothelial cells, but also regulate vascular differentiation, regression, and permeability. During and after cerebral ischemia, brain vasculature becomes leaky and unstable, and the normally impermeable blood-brain barrier breaks down. Several days after the ischemic insult, endothelial cells begin to proliferate, and angiogenesis occurs. Expression studies have shown that key vascular GFs are regulated, during these processes, in a complex and coordinated manner. The distinct pattern of regulation exhibited by each vascular GF suggests a unique role for each factor during the initial vascular destabilization and subsequent angiogenesis that occurs after cerebral ischemia. Data from studies in other biological systems support these suggested roles. Thus, manipulation of vascular GFs may prove to be an effective means of stabilizing or enriching brain vasculature after ischemia, and ameliorating the detrimental effects of blood-brain barrier breakdown and vessel regression after stroke.     

Characteristics of tumour vessels in cytological squash smears of astrocytic tumours

S. Sato, Y. Sato, K. Marutsuka, H. Takeshima and Y. Asada Characteristics of tumour vessels in cytological squash smears of astrocytic tumours Objective: Smear preparations are useful tools from which to diagnose brain tumours intraoperatively. Although vascular proliferation is histologically a key feature of high‐grade astrocytoma, the characteristics of tumour vessels in smear preparations have not been determined. Methods: We examined the density and morphological parameters (area, width, nuclear layer and branches of vessel wall) of tumour vessels in squash smears of 43 primary astrocytomas (grade II diffuse astrocytomas, n = 9; grade III anaplastic astrocytomas, n = 13; grade IV glioblastomas, n = 21) and normal brain tissues (n = 11). Results: Vessel density and all morphological parameters were significantly higher in grade IV than in the other grades of tumours and in normal brain tissue. Vessel area, width and nuclear layer were greater in grade III than in normal brain tissue. The sensitivity and specificity of these vessel parameters for astrocytomas were 75–100% and 82–100%, respectively. Conclusions: Tumour vessel evaluations from squash smears provide useful information for the intraoperative diagnosis and grading of astrocytic tumours.