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M N Ghabriel

Publications and source records attributed to M N Ghabriel.

At least 19 recordsLinked to original sources
PubMed1996

Dural microvessels: molecular properties of their luminal anionic sites.

(1) Neurogenic inflammation has been implicated in the pathogenesis of the vascular headaches of migraine and cluster headaches. (2) Dural blood vessels are both pain-sensitive and show neurogenic plasma extravasation. (3) Endothelial cell (EC) surface anionic sites appear to be a determinant of vascular permeability. We therefore examined the anionic sites of dural EC to determine whether they are different from those of pial and parenchymal vessels. Luminal anionic sites of rat optic nerve EC were labelled with cationic colloidal gold (CCG) and cationic ferritin (CF) and examined by electron microscopy. Employing a battery of enzymes, the effects of digestion of ultrathin sections on subsequent labelling with CCG was quantified using image analysis software. In addition, a gold-labelled lectin, wheat-germ agglutinin (WGA), was employed to locate specific saccharide residues. Of the enzymes with a narrow specificity, only neuraminidase substantially reduced CCG binding. Of the proteolytic enzymes, papain was most effective in reducing labelling. These results show that the luminal EC anionic sites are chiefly composed of sialoglycoproteins. The labelling with biotinylated WGA-streptavidin gold was similar to that with CCG without enzyme digestion. This suggests that WGA is binding to N-acetylneuraminic (sialic) acid residues and not to the neutral N-acetylglucosamine (since CCG would not label uncharged molecules). These results do not differ significantly from those for pial and parenchymal EC. It is therefore likely that factors other than anionic site molecular composition account for the susceptibility of dural vessels to neurogenic plasma extravasation. The relevance of these observations in an experimental animal model to the human clinical condition remains to be determined.

Animals↗
PubMed1995

Optic nerve microvessels: a partial molecular definition of cell surface anionic sites.

Incorporated in the luminal glycocalyx of vascular endothelia (EC) are negatively charged microdomains (anionic sites). These sites are considered functionally important (a) in their interaction with circulating blood constituents, and (b) as a determinant of vascular permeability. The molecular composition of these EC sites, described for a number of tissues, has demonstrated a heterogeneity dependent on their anatomical location. Luminal anionic sites have not been characterized for EC of optic nerve. Optic nerves were removed from Sprague-Dawley rats previously fixed by vascular perfusion. EC anionic sites were labelled with the probes cationic colloidal gold (CCG) and cationic ferritin (CF), using the pre- and post-embedding techniques, and examined by electron microscopy. The effects of enzyme digestion of ultrathin sections on subsequent CCG labelling were determined using a battery of enzymes in association with the post-embedding technique. CCG labelling was quantified following each enzyme treatment using image analysis software. The biotinylated lectin wheat germ agglutinin (WGA) with streptavidin gold was also used to localize specific monosaccharide residues. The luminal front of intraneural EC showed a uniform labelling with CCG and CF which was greater than on the abluminal surface. Extracellular matrix components and basal laminae were moderately labelled. Digestion of tissue sections with heparitinase and trypsin had no significant effect on subsequent CCG labelling. Proteinase K was less effective than papain but both produced a significant reduction. Neuraminidase almost completely eliminated labelling. CCG binding to the luminal plasma membrane of optic nerve EC can be significantly reduced with proteolytic and glycolytic enzymes. The results demonstrate that sialoglycoproteins principally constitute these luminal EC anionic sites. Biotinylated WGA-streptavidin gold, which detects both N-acetylneuraminic (sialic) acid and N-acetylglucosamine, gave a similar pattern of labelling to CCG alone on the luminal versus abluminal EC fronts. These findings suggest that WGA is binding predominantly to N-acetylneuraminic acid residues since CCG would not label the neutral (uncharged) N-acetylglucosamine.

Animals↗
PubMed1995

Distribution of a putative endothelial barrier antigen in the ocular and orbital tissues of the rat.

BACKGROUND: A rat endothelial barrier antigen (EBA) recognised by a monoclonal antibody has been shown to be expressed strongly by endothelial cells of brain capillaries possessing a blood-brain barrier and only weakly expressed by fenestrated brain vessels. METHODS: In this study immunocytochemical methods for light and electron microscopy were used to study EBA distribution in the eye and orbital tissues of the rat. RESULTS: Blood-ocular barrier vessels in the optic nerve, retina, iris, and some vessels in th choroid and ciliary body were immunopositive for EBA. By pre-embedding immunocytochemistry for electron microscopy the antigen was observed on the luminal endothelial cell surface. CONCLUSION: Surprisingly, some non-barrier vessels in the ciliary body and choroid expressed EBA suggesting that it may play a broader role in endothelial properties than previously recognised. The functional significance of EBA remains to be elucidated.

Animals↗
PubMed1995

Differential expression of an endothelial barrier antigen between the CNS and the PNS.

A monoclonal antibody to an antigen (EBA) expressed by neural endothelial cells (EC) was used to investigate any difference in the distribution of EBA between the CNS and PNS. Pre-embedding ultrastructural cytochemistry of rat sciatic and optic nerves was undertaken using anti-EBA, detected with a silver-enhanced gold-conjugated secondary antibody. LM immunocytochemical localisation of EBA was also performed using an HRP-conjugated secondary antibody. EC of pial and parenchymal optic nerve vessels were strongly immunopositive for EBA. Vessels of the dura were negative. At the EM level EBA was observed on the EC luminal surface. In contrast, EC of sciatic nerve were either negative or only weakly immunopositive. The molecular characteristics and function of EBA are largely unknown. Therefore the functional significance of the present findings remains to be determined.

Animals↗
PubMed1992

Influence of fatty acid content of lysophosphatidyl choline on its myelinotoxic properties.

The efficacy of lysophosphatidyl choline (LPC) type I and type IV in producing demyelination was assessed in rat tibial and sural nerve. By light and electron microscopy, a greater myelinolytic activity was demonstrated with type I, and concomitantly electrophysiology showed a more severe conduction block. In teased nerve preparations and 1-microns thin sections, demyelinated fibres were more frequent with LPC type I. At 1 h after injection, electron microscopy showed much more extensive myelin lysis in the form of fine vesicular debris. By 6 days, completely demyelinated fibres were much more common and associated Schwann cells contained either small quantities or no myelin debris. With type IV LPC, cytopathological changes were more extensive at 1 h. A minority of Schwann cells showed swollen hydropic cytoplasm and degradation of organelles. Axonal retraction from the myelin sheath occurred in occasional fibres, and in a few unmyelinated fibres axoplasm showed organelle depletion and increased granularity. By 6 days, Schwann cells still contained large quantities of gross myelin debris and had often retracted to expose extensive areas of axolemma. The findings suggest that the two types of LPC have different myelinolytic actions, which may be related to their different fatty acid content. A possible role for the two types of LPC in 'bystander demyelination' is considered.

Animals↗
PubMed1989

Diffusion barrier properties of the perineurium: an in vivo ionic lanthanum tracer study.

While the perineurium as a diffusion barrier has been extensively investigated by light and electron microscopy, such studies have been largely restricted to the use of protein tracers. In the present study the permeability of the perineurium to a physiologically more relevant ionic tracer has been assessed. In vivo the rat sural or tibial nerve was either microinjected with lanthanum nitrate solution for endoneurial application or bathed in the lanthanum solution for epineurial application. The findings generally demonstrated an effective barrier to the tracer which failed to penetrate the inner layers of the perineurium. Only at the highest lanthanum concentration and longest time intervals employed did trace quantities occasionally penetrate the barrier and then only in the presence of some cytopathological changes to the outermost perineurial cells. The usefulness of the microinjection method was limited by the slight but unavoidable trauma to the perineurium. The findings are related to those of other studies which have used electron dense tracers, also to studies using physiological including electrophysiological techniques and morphological including freeze-fracture methods.

Animals↗
PubMed1989

Lysophosphatidylcholine-induced incipient demyelination: involvement of a new tubular structure.

Demyelination was induced in the rat sciatic and tibial nerves by microinjection with lysophosphatidylcholine (LPC). Accompanying early myelin lysis (1-24 h) was the formation of vesicles and tubular structures. The tubules which are novel structures have a diameter range of 24-27 nm, a centre-to-centre spacing 30-50 nm and may extend for 3 microns in length. In this form they are arranged as a monolayer in the periaxonal space. As demyelination progressed and the periaxonal space widened the tubules increased in number and became more irregularly arranged. The tubules are apparently derived from the myelin lamellae/Schwann cell plasma membrane, while the axolemma remains intact.

Animals↗
PubMed1988

Lysophosphatidyl choline-induced demyelination. A freeze-fracture study.

Focal demyelination was produced in the rat sciatic nerve by microinjection of lysophosphatidyl choline (LPC). The demyelinating lesion was examined over the following 48 h using the freeze-fracture technique to examine myelin, Schwann cell and axonal membranes. Myelin lamellae were replaced by myriad spherical or oval membranous vesicles. The axonal and Schwann cell plasma membranes remained intact and the latter showed a large increase in caveolae-associated pores in some nerve fibres. The lysis of myelin lamellae and membranous vesicle formation are related to the known action of LPC on myelin and its membrane fusogenic properties. The importance of calcium ion influx and membrane protein aggregation and depletion in vesiculation are discussed.

Animals↗
PubMed1987

The blood-nerve barrier: an in vivo lanthanum tracer study.

The permeability of the blood-nerve barrier was investigated using ionic lanthanum as an electron-dense tracer. The rat sural nerve was microinjected in vivo with lanthanum nitrate solution either into the endoneurial space or into the epineurium. Five to sixty minutes after injection the sural nerves were fixed by vascular perfusion or immersion. Using electron microscopy, lanthanum tracer was observed to be associated with endoneurial vessels in the perivascular spaces, in the inter-endothelial clefts and within the lumina. Furthermore, tracer was present in the spaces between adjacent endothelial cell layers and within vesicles and caveolae of endothelial cells. Epineurial vessels showed a similar distribution of tracer deposits but in greater quantities in inter-endothelial cell spaces and vessel lumina. The results are considered to demonstrate an absence of a blood-nerve barrier to ions as exemplified by lanthanum and are compatible with data from physiological experiments. The blood-nerve and blood-brain barriers are contrasted in their permeability to ions, their related fine structure and their physiological roles.

Animals↗
PubMed1987

The development of Schmidt-Lanterman incisures: an electron microscope study.

The development of Schmidt-Lanterman incisures was investigated in the rat sural nerve during an active phase of postnatal myelination (5-21 days post partum). Two distinct populations of incisures were recognised and the following nomenclature for their developmental stages is proposed. Primary incisures which appear ab initio in myelination and always extend across the whole radial thickness of the myelin sheath but initially around only part of its circumference. Consequently they appear in transverse section as sectoral incisures (occupying less than half the circumference) and in longitudinal section as asymmetric incisures (involving one side only of the myelin sheath). Secondary incisures appear later, in regions of a compact myelin sheath, initially traversing only part of its radial thickness but commonly occupying its whole circumference. Thus they usually appear in transverse section as circumferential incisures and in longitudinal section as symmetric incisures (involving both sides of the myelin sheath). Less commonly secondary incisures may form in a sector of the myelin sheath but still in regions of compact myelin and thus appear asymmetric in longitudinal section and sectoral in transverse section. Secondary incisures appear mainly adaxonally in the earlier stages examined and mainly abaxonally in the later stages. The maturation of primary and secondary incisures into the radially and circumferentially complete incisure characteristic of the mature myelinated nerve fibre is described. The above mechanisms of incisural formation are contrasted with mechanisms previously suggested to occur during normal development and remyelination and related to the plasticity and ultrastructure of the myelin sheath.

Animals↗
PubMed1985

Axonal microtubules: a computer-linked quantitative analysis.

Employing current computer-aided morphometric techniques, axonal microtubule density was determined for the rat sural nerve. Analysis of extensive data showed that while microtubule number increases with axon size, the increase is not directly proportional. Thus the relationship between microtubule density and axonal size is inversely related, so that microtubule density is greater in smaller axons than in larger axons. When a proximal and distal site, separated by 2 cm, were compared for microtubule density there was no significant difference, using pooled data for all fibre diameters. The results are interpreted in terms of our present knowledge of axonal-microtubule quantitative relationships, which is reviewed.

Animals↗
PubMed1985

Axonal degeneration in large and small nerve fibres. An electron-microscopic and morphometric study.

Using computer-aided morphometric methods, axonal degeneration following nerve crush was analysed to reassess whether small fibres degenerate before large fibres or vice versa, or simultaneously. Axonal microtubule density was used as the criterion for determining the extent of fibre degeneration. Axonal areas and axonal microtubule numbers were recorded from a large sample of myelinated fibres in the right unoperated rat sural nerve and distal to crush in the left sural nerve. Both samples were divided into small and large fibre groups, according to axonal areas. Statistical analysis of the data confirmed a significant loss of microtubules from the left crushed nerve fibres but no significant difference in the relative loss of microtubules from small and large fibres. It is concluded, therefore, that in Wallerian degeneration, axonal breakdown, as assessed by microtubule loss, occurs simultaneously in small and large fibres. The findings are related to the electrophysiological changes which occur in Wallerian degeneration.

Animals↗
PubMed1984

Nodes of Ranvier and Schmidt-Lanterman incisures: an in vivo lanthanum tracer study.

The permeability of the tight junctional system of myelin, at the juxtanodal myelin terminal loops and Schmidt-Lanterman incisures, was investigated using the ionic tracer lanthanum (a) in vivo followed by fixation, (b) concurrently with fixation, (c) following fixation. Employing the same methods the juxtanodal membrane complex formed between myelin loops and axolemma was also tested. The results of this study demonstrate that the periaxonal space (between axon and Schwann cell) is apparently accessible to lanthanum via the myelin loop-axolemmal junction, irrespective of the mode of exposure of myelinated fibres to the tracer. Similarly, the tight junctions between adjacent myelin terminal loops apparently do not prevent lanthanum penetration either in living or in fixed nerves. By contrast the tracer obtained access to the extracellular space within incisures only in vivo. The results are interpreted in terms of the permeability of nodes and incisures in vivo to physiologically important ions and related to current concepts of the electrophysiology of the myelinated nerve fibre.

Animals↗
PubMed1984

Myelinated nerve fibres and the fate of lanthanum tracer: an in vivo study.

The permeability of the marginal tight junctional system of myelin was tested in the rat employing the electron-dense tracer lanthanum nitrate. Lanthanum was either included in the fixative used for vascular perfusion (at a concentration of 20 mM) or was microinjected in vivo into the sural or tibial nerve (5, 10 and 20 mM). After 5-60 minutes, the microinjected nerves were fixed either by immersion or vascular perfusion. Lanthanum tracer was present in the intraperiod line gap of myelin, irrespective of the mode of application of the tracer, the method of fixation or the time of exposure to lanthanum. However, the tracer was present more extensively when included in the fixative compared with in vivo microinjection. Internodally, lanthanum was usually restricted to the inner, or more commonly, the outer lamellae of larger fibres, while all lamellae were usually penetrated by tracer in smaller fibres. Paranodally, compact myelin was more extensively penetrated. The periaxonal space (between axon and Schwann cell) was readily accessible to tracer. It is concluded that the marginal tight junctional system of myelin is apparently of the 'leaky' type and is permeable to ions. The findings have implications for the electrophysiology and pathophysiology of the myelinated nerve fibre.

Animals↗
PubMed1984

The non-directional pattern of axonal changes in Wallerian degeneration: a computer-aided morphometric analysis.

Wallerian degeneration was investigated to determine whether axonal changes occur progressively in a somatofugal or somatopetal direction or simultaneously along the length of the axon. Microtubule density was used as a measure of the extent of axonal degeneration and was assessed by a computer-aided analysis of electron micrographs. The left sural nerves of ten rats were crushed and 30 hours later axonal areas and axonal microtubule numbers were recorded from a large sample of axons at two sites 1 cm and 3 cm distal to the crush. The same recordings were made from the right unoperated nerve at two comparable sites. Statistical analysis of all the data provided no evidence for a somatofugal or reverse direction of degeneration. It is concluded therefore that in Wallerian degeneration axonal changes, as indicated by microtubule dissolution, occur simultaneously along the length of the axon. It is proposed that to interpret the conflicting published data on the direction of fibre degeneration, Schwann cell changes (e.g. myelin ovoid formation) and axonal changes (e.g. microtubule dissolution) should be considered independently since they have different aetiological mechanisms which may account for the differing experimental results reported.

Animals↗
PubMed1983

A combined morphological and electrophysiological study of conduction block in peripheral nerve.

The reliability of the electrophysiological criterion of conduction block in determining the presence of focal demyelination in a peripheral nerve has been studied in an animal model. Demyelination was produced in the rat tibial nerve by one or two closely spaced microinjections of lysophosphatidylcholine (LPC). Histological and electrophysiological data were obtained on the acute lesion (up to 6 days), and during recovery (up to 11 weeks). Single LPC injections produced a lesion of very variable severity. Double injections more reliably produced a severe lesion with marked conduction block. Slight axonal damage was occasionally seen in nerves showing severe demyelination. The ratio of amplitude of muscle action potentials evoked by stimuli proximal and distal to the sites of nerve injection was calculated to detect the development of conduction block. The post injection ratio was more than 2 standard deviations below the control mean in 86% of nerves showing signs of demyelination. No control saline injected nerves showed such evidence of conduction block. The severity of the electrophysiological abnormality did not prove a reliable indicator of the severity of the histological lesion, however. The possible reasons for this variability are discussed and it is argued that caution should be exercised when interpreting this particular electrophysiological finding in clinical practice.

Animals↗
PubMed1982

Demyelination: a failure of cell communication?

A hypothesis is proposed that demyelination in both the CNS and PNS involves a failure of cell communication between the axon and oligodendrocyte/Schwann cell, as a primary event. The site of communication is assumed to be the paranodal myelin loop-axolemma membrane complex. It is postulated that "cross-talk" between the two cell types can be interrupted, and hence demyelination initiated, by pathophysiological changes in either the axon or myelinating cell. Experimental evidence in support of the hypothesis is cited in so far as it exists.

Axonal Transport↗