Capillaries In Central Nervous System Research Articles

As Verified with the Aid of Biotinylated Spermine, the Brain Cannot Take up Polyamines from the Bloodstream Leaving It Solely Dependent on Local Biosynthesis.

The importance of polyamines (PAs) for the central nervous system (CNS) is well known. Less clear, however, is where PAs in the brain are derived from. Principally, there are three possibilities: (i) intake by nutrition, release into the bloodstream, and subsequent uptake from CNS capillaries, (ii) production by parenchymatous organs, such as the liver, and again uptake from CNS capillaries, and (iii) uptake of precursors, such as arginine, from the blood and subsequent local biosynthesis of PAs within the CNS. The present investigation aimed to unequivocally answer the question of whether PAs, especially the higher ones like spermidine (SPD) and spermine (SPM), can or cannot be taken up into the brain from the bloodstream. For this purpose, a biotin-labelled analogue of spermine (B-X-SPM) was synthesized, characterized, and used to visualize its uptake into brain cells following application to acute brain slices, to the intraventricular space, or to the bloodstream. In acute brain slices there is strong uptake of B-X-SPM into protoplasmic and none in fibrous-type astrocytes. It is also taken up by neurons but to a lesser degree. Under in vivo conditions, astrocyte uptake of B-X-SPM from the brain interstitial fluid is also intense after intraventricular application. In contrast, following intracardial injection, there is no uptake from the bloodstream, indicating that the brain is completely dependent on the local synthesis of polyamines.

A novel functional role for the classic CNS neurotransmitters, GABA, glycine, and glutamate, in the kidney: potent and opposing regulators of the renal vasculature.

The presence of a renal GABA/glutamate system has previously been described; however, its functional significance in the kidney remains undefined. We hypothesized, given its extensive presence in the kidney, that activation of this GABA/glutamate system would elicit a vasoactive response from the renal microvessels. The functional data here demonstrate, for the first time, that activation of endogenous GABA and glutamate receptors in the kidney significantly alters microvessel diameter with important implications for influencing renal blood flow. Renal blood flow is regulated in both the renal cortical and medullary microcirculatory beds via diverse signaling pathways. GABA- and glutamate-mediated effects on renal capillaries are strikingly similar to those central to the regulation of central nervous system capillaries, that is, exposing renal tissue to physiological concentrations of GABA, glutamate, and glycine led to alterations in the way that contractile cells, pericytes, and smooth muscle cells, regulate microvessel diameter in the kidney. Since dysregulated renal blood flow is linked to chronic renal disease, alterations in the renal GABA/glutamate system, possibly through prescription drugs, could significantly impact long-term kidney function.NEW & NOTEWORTHY Functional data here offer novel insight into the vasoactive activity of the renal GABA/glutamate system. These data show that activation of endogenous GABA and glutamate receptors in the kidney significantly alters microvessel diameter. Furthermore, the results show that these antiepileptic drugs are as potentially challenging to the kidney as nonsteroidal anti-inflammatory drugs.

Piezo1 Is a Mechanosensor Channel in Central Nervous System Capillaries.

Capillaries are equipped to sense neurovascular coupling agents released onto the outer wall of a capillary, translating these external signals into electrical/Ca2+ changes that play a crucial role in blood flow regulation and ensuring that neuronal demands are met. However, control mechanisms attributable to forces imposed onto the lumen are less clear. Here, we show that Piezo1 channels act as mechanosensors in central nervous system capillaries. Electrophysiological analyses confirmed expression and function of Piezo1 channels in brain cortical and retinal capillaries. Activation of Piezo1 channels evoked currents that were sensitive to endothelial cell-specific Piezo1 deletion. Using genetically encoded Ca2+ indicator mice and an ex vivo pressurized retina preparation, we found that activation of Piezo1 channels by mechanical forces triggered Ca2+ signals in capillary endothelial cells. Collectively, these findings indicate that Piezo1 channels are capillary mechanosensors that initiate crucial Ca2+ signals and could, therefore, have a profound impact on central nervous system blood flow control.

Pericyte-to-endothelial cell signaling via vitronectin-integrin regulates blood-CNS barrier

Endothelial cells of blood vessels of the central nervous system (CNS) constitute blood-CNS barriers. Barrier properties are not intrinsic to these cells; rather they are induced and maintained by CNS microenvironment. Notably, the abluminal surfaces of CNS capillaries are ensheathed by pericytes and astrocytes. However, extrinsic factors from these perivascular cells that regulate barrier integrity are largely unknown. Here, we establish vitronectin, an extracellular matrix protein secreted by CNS pericytes, as a regulator of blood-CNS barrier function via interactions with its integrin receptor, α5, in endothelial cells. Genetic ablation of vitronectin or mutating vitronectin to prevent integrin binding, as well as endothelial-specific deletion of integrin α5, causes barrier leakage in mice. Furthermore, vitronectin-integrin α5 signaling maintains barrier integrity by actively inhibiting transcytosis in endothelial cells. These results demonstrate that signaling from perivascular cells to endothelial cells via ligand-receptor interactions is a key mechanism to regulate barrier permeability.

Endothelial Cells Filopodia in the Anastomosis of Central Nervous System Capillaries.

In this article we explore filopodia of endothelial cells (ECs) in the developing central nervous system (CNS) using the Golgi method and transmission electron microscopy. Filopodia of ECs play a crucial role in the anastomosis of growing capillaries of the CNS. The leading ECs filopodia from approaching capillaries interconnect forming complex conglomerates that precede the anastomotic event. The contacting filopodia form narrow spaces between them filled with proteinaceous basal lamina material. The original narrow spaces coalesce into larger ones leading to the formation of a single one that will interconnect (anastomose) the two approaching capillaries. The four leading ECs (two for each approaching capillary) become the wall of the newly formed post-anastomotic CNS capillaries. These new CNS capillaries are very small with narrow and irregular lumina that might permit the passage of fluid but not yet of blood cells. Eventually, their lumen enlarges and permits the passage of blood cells.

Bidirectional control of CNS capillary diameter by pericytes

Neural activity increases local blood flow in the central nervous system (CNS), which is the basis of BOLD (blood oxygen level dependent) and PET (positron emission tomography) functional imaging techniques. Blood flow is assumed to be regulated by precapillary arterioles, because capillaries lack smooth muscle. However, most (65%) noradrenergic innervation of CNS blood vessels terminates near capillaries rather than arterioles, and in muscle and brain a dilatory signal propagates from vessels near metabolically active cells to precapillary arterioles, suggesting that blood flow control is initiated in capillaries. Pericytes, which are apposed to CNS capillaries and contain contractile proteins, could initiate such signalling. Here we show that pericytes can control capillary diameter in whole retina and cerebellar slices. Electrical stimulation of retinal pericytes evoked a localized capillary constriction, which propagated at approximately 2 microm s(-1) to constrict distant pericytes. Superfused ATP in retina or noradrenaline in cerebellum resulted in constriction of capillaries by pericytes, and glutamate reversed the constriction produced by noradrenaline. Electrical stimulation or puffing GABA (gamma-amino butyric acid) receptor blockers in the inner retina also evoked pericyte constriction. In simulated ischaemia, some pericytes constricted capillaries. Pericytes are probably modulators of blood flow in response to changes in neural activity, which may contribute to functional imaging signals and to CNS vascular disease.

Blood-Spinal Cord and Brain Barriers in Health and Disease Edited by: Hari Shanker Sharma, Jan Westerman. Elsevier Academic Press, San Diego, CA; 2004.

The health of the central nervous system (CNS) is a subject that impacts on all people and especially on those who have the misfortune to be afflicted by CNS injury or disease. The cerebrovascular barriers at the CNS capillaries and the choroid plexus play a vital role in maintaining the microenvironment of the cells in the brain and spinal cord. The stated aim of this book is to act as a stimulus for research directed towards minimizing the effects of CNS stress, injuries and insults. With seven sections that include 27 chapters from 59 contributors and 605 pages in total, this book covers many topics including: basic aspects of the barriers, in vitro models, physiological transport mechanisms, neurochemical mediators, stress situations, changes in disease conditions and clinical aspects. Each chapter consists of a review, together with up-to-date original data. The chapters are laid out clearly in a consistent format with informative illustrations. This is an important wide-ranging book aimed at researchers in neuroscience, drug discovery and development, and neuropathology. Although large, it contains a wealth of information on contemporary research with an impressive 29-page index from arachadonic acid to zonula occludens proteins. The bulk of the text deals with research relating to the cerebrovascular barriers in the brain. However, in contrast with past books on the blood-brain barriers, this one contains several chapters concerned with the blood-spinal cord barrier. Considering the volume of research on spinal cord injury in recent decades, this serves as a useful source of information and a reminder that barrier function is important throughout the CNS. Furthermore, it reinforces the probability that information obtained from research on barriers in the brain can be applied to the therapeutic treatment of spinal cord injury. One rather disappointing aspect of this otherwise excellent book is the small amount of space devoted to the choroid plexus and the cerebrospinal fluid (CSF). This is the alternative route to the brain, the blood-CSF barrier with somewhat different properties to the blood-brain barrier. Only two chapters contribute to this very important aspect of brain homeostasis and to the potential for manipulation of the CSF for diagnostic and therapeutic purposes. Overall, 'Blood-Spinal Cord and Brain Barriers in Health and Disease' is a contemporary and very informative volume that should be a ready source of reference for all researchers and clinicians concerned with the CNS in health and disease.

Elimination of phenolsulfonphthalein from the cerebrospinal fluid via capillaries in central nervous system in cats by active transport

It was recently proposed that organic anions, such as cerebral acidic metabolites and phenolsulfonphthalein (PSP), are eliminated from cerebrospinal fluid (CSF) by diffusion into the central nervous system (CNS) and further by active transport into capillaries. To test this hypothesis, PSP was injected into cisternal CSF and its distribution into various parts of the CNS was measured 1 and 3 h later in control cats and those pretreated with probenecid, which blocks active transport of organic anions into capillaries. PSP in tissue shows an intensive pink color when exposed to 1 N NaOH. Planimetric analysis of color pictures of coronal CNS slices showed that at the first hour, diffusion and distribution of PSP into the CNS in both groups of animals was similar, while at the third hour, a great reduction of PSP distribution in the CNS in control and only a slight reduction in probenecid pretreated cats was observed. The results support the hypothesis that active transport across the capillary wall in the CNS is the main avenue for elimination of cerebral acidic metabolites from both CSF and CNS and in such a way that central homeostasis is maintained.

A non-invasive transport system for GDNF across the blood-brain barrier.

Glial cell line-derived neurotrophic factor (GDNF) is a neurotrophin which supports midbrain dopaminergic neurons and spinal cord motorneurons. GDNF has been proposed as a possible therapeutic agent for Parkinson's disease, spinal cord injury or motorneuron degenerative disorders. Administration of GDNF is complicated by its poor penetration across the blood-brain barrier (BBB). Central nervous system capillaries are uniquely enriched in transferrin receptors and antibodies to these receptors (OX-26) have been proposed as potential carriers to transport large molecules across the BBB. Intravenous administration of an OX-26-GDNF conjugate enhanced survival of spinal cord motorneurons in intraocular transplants, which possess an organotypic BBB. This suggests that the OX-26-GDNF conjugate could be utilized for non-invasive treatment of neurodegenerative diseases of the spinal cord or midbrain dopaminergic neurons.

Purified Shiga-like toxins induce expression of proinflammatory cytokines from murine peritoneal macrophages.

Infections with Shiga toxin-producing Shigella dysenteriae type 1 and Shiga-like toxin (SLT)-producing Escherichia coli cause outbreaks of bloody diarrhea in which patients are at risk for developing life-threatening complications involving the renal and central nervous systems. Histopathology studies and in vitro experiments suggested that the toxins damage toxin receptor-expressing endothelial cells (EC) lining glomerular and central nervous system capillaries. In the presence of inducible host factors (cytokines), EC sensitivity to SLT toxicity was increased approximately 1 million-fold. We hypothesized that to manifest the vascular lesions characteristic of infection with toxin-producing bacteria, two signals were needed: systemic toxins and elevated proinflammatory cytokines (tumor necrosis factor alpha [TNF-alpha], interleukin 1 [IL-1], and IL-6). Human EC do not secrete these cytokines when stimulated with SLTs in vitro, suggesting that additional cells may be involved in pathogenesis. Therefore, we carried out comparative analyses of the capacity of purified (endotoxin-free) SLTs and lipopolysaccharides (LPS) to induce cytokine mRNA and proteins from murine macrophages. The cells were essentially refractory to SLT cytotoxicity, expressing low to undetectable levels of toxin receptor. SLTs and LPS induced TNF activity and IL-6 expression from macrophages, although dose response and kinetics of cytokine induction differed. LPS was a more effective inducing agent than SLTs. SLT-I-induced TNF activity and IL-6 expression were delayed compared with induction mediated by LPS. IL-1 alpha production required approximately 24 h of exposure to SLTs or LPS. Macrophages from LPS-hyporesponsive C3H/HeJ mice produced low levels of TNF activity when treated with SLT-I, suggesting that LPS and SLTs may utilize separate signaling pathways for cytokine induction.

Morphological observations on the unique paired capillaries of the opossum retina.

Light-microscopic and ultrastructural analysis of the ocular tissues of the North American opossum (Didelphis virginiana) revealed that the arterial and venous segments of retinal vessels, including capillaries of the smallest calibre, occur in pairs. They do not form anastomotic networks, the common pattern in mammals with vascularised retinae, but instead the two segments of the pair join to form hairpin end loops. The paired vessels, with the arteriolar limb usually on the vitread aspect, penetrate the retina and branch to form three distinct layers of capillaries. The most superficial lies in the nerve fiber layer, the middle is situated in the inner nuclear layer and the deepest extends to the external limiting membrane, which is considerably deeper than in normal mammalian holangiotic retinae. The paired capillaries display classical morphological features of central nervous system capillaries, i.e., they are lined by continuous endothelial cells united by tight junctions. The lining endothelium is supported by a distinct basal lamina that splits to envelop pericytes. The latter, although abundant, are invariably interposed between the two vessels that form each vascular unit. Phylogenetic and functional aspects of this unique form of retinal vascularisation are discussed.

A study of the expression of laminin in the spinal cord of the frog during development and regeneration.

In the present experiments, we have used an affinity-purified polyclonal antibody to laminin to determine the time course of expression of laminin in the central nervous system (CNS) of Rana temporaria tadpoles during normal development and during restoration of the dorsal columns of the spinal cord. Immunoblotting analysis indicated that in the peripheral nervous system (PNS) of adult frogs the antibody recognized proteins of molecular weights 350-400 and 205-220 kDa, corresponding to the A and B chains respectively of mammalian laminin. Immunohistochemistry with our antibody suggested that laminin was absent from the tadpole spinal cord, and did not appear even in the basal lamina of the blood vessels within the spinal cord until after metamorphosis. Furthermore, there was no evidence of laminin expression in the dorsal columns after hemisection of the spinal cord. However, throughout development laminin was present in basal lamina outside the CNS, in particular in the pial membranes, and in the basal lamina of blood vessels and sheath cells in the dorsal and ventral roots. Electron microscopy showed that the blood vessels of CNS capillaries had basal laminae throughout development that was morphologically indistinguishable from that seen in peripheral vessels.

Basement membrane of central nervous system capillaries lacks ruthenium red-staining sites

We used the cationic dye ruthenium red to examine the distribution of anionic macromolecules (presumably proteoglycans) in the basement membranes of the fenestrated capillaries and epithelium of the choroid plexus, and of the continuous capillaries forming the blood-brain barrier. Both the endothelial and epithelial basement membranes of choroid plexus displayed discrete, 10- to 20-nm-diameter, electron-dense sites after exposure to ruthenium red. These sites were similar in size, appearance, and distribution to those found in the vascular and epithelial basement membranes of a variety of tissues outside the central nervous system. In contrast, the basement membranes of continuous, blood-brain barrier capillaries did not display electron-dense sites following exposure to ruthenium red, even after measures had been taken to enhance penetration of the dye across the endothelial cells. The lack of discrete ruthenium red staining in the basement membrane of continuous blood-brain barrier capillaries could be due to a relative paucity of anionic macromolecules, or may be the result of the compact architecture of this particular basement membrane. Regardless of the final explanation, these findings suggest that the basement membrane of the blood-brain barrier, like its endothelium, is structurally (and perhaps functionally) unique.

Cellular aspects of transcapillary exchange.

Mise au point bibliographique sur l'organisation de la paroi capillaire et de la membrane des cellules de ces capillaires ainsi que sur les processus d'endocytose et de transcytose

The pathway between hyaloid blood and retinal neurons in the toad. Structural observations and permeability to tracer substances.

The hyaloid vessels form a capillary network on the inner surface of the retina. These capillaries are embedded in the vitreous humor, and they lack a glial investment. The intercellular spaces of the retina communicate with the ocular cavity, as can be evidenced by following the penetration of tracer substances. Hence, there is an extracellular diffusion pathway between hyaloid capillaries and retinal neurons, without interposition of glial cells. Trypan blue and ferrocyanide were not detected within the vitreous humor nor the retina after systemic injection. To this extent, at least, the hyaloid capillaries functionally resemble central nervous system capillaries. Intravascular injections of horseradish peroxidase established the absence of vesicular transfer across the endothelium of the hyaloid capillaries. In addition, quintuple-layered junctions between endothelial cells prevented the intercellular passage of the enzyme. It is likely, therefore, that the only pathway across the endothelium of the hyaloid capillaries is through the plasmalemma of the endothelial cells.