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Biomedical subjects

N J Lane

Publications and source records attributed to N J Lane.

At least 19 recordsLinked to original sources
PubMed1992

Ebselen. Antioxidant capacity in renal preservation.

Ebselen (PZ51) was tested for its ability to inhibit oxidative membrane damage and improve outcome of rabbit kidneys rendered cold ischaemic for 72 hr. In view of the rapid metabolism of ebselen, the antioxidant capacities of its two principal metabolites were first compared with that of the parent drug in an in vitro hepatic microsomal lipid peroxidation system initiated by NADPH/Fe(3+)-ADP. The potent antioxidant activity of ebselen was confirmed but metabolite I (2-glucuronylselenobenzanilide) exhibited no antioxidant potential up to a concentration of 50 microM; metabolite II (4-hydroxy-2-methyl-selenobenzanilide) did inhibit lipid peroxidation but was about 80 times less effective than the parent compound. The storage of rabbit kidneys in hypertonic citrate solution at 0 degrees for 72 hr of cold ischaemia resulted in greatly increased susceptibility to oxidative membrane damage in both the cortex and medulla as determined by the subsequent in vitro formation of two markers of lipid peroxidation (Schiff's bases and thiobarbituric acid-reactive material). Inclusion of ebselen (50 microM) in the flush and storage solution led to a highly significant reduction in these oxidative markers in both regions of the kidney. Intracellular and interstitial oedema was noted in organs subjected to 72 hr cold ischaemia and was reduced by ebselen (50 microM in the flush/storage solution). The rate of post-ischaemic lipid peroxidation was found to correlate well with the extent of oedema in the renal medulla (r = 0.84, P less than 0.001) but no such correlation was found in the cortex. Administration of ebselen (5.5 mg/kg i.v. and 100 microM in the flush/storage solution) did not improve the long-term survival of rabbits following autotransplantation of a single kidney stored for 48 or 72 hr. No protective effect of ebselen could be demonstrated either in terms of graded physiological function or histological outcome.

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PubMed1992

Freeze-fracture evidence for a novel restricting junction at the blood-brain barrier of the cuttlefish Sepia officinalis.

The blood-brain barrier in the cuttlefish Sepia officinalis has been studied with the freeze-fracture technique. Previous thin-section electron microscopy showed that a restricting junction is formed between perivascular glial processes in microvessels and venous vessels, and between pericytes in arterial vessels; the restriction appeared not to be a classical zonula occludens or septate junction. In freeze-fracture replicas from brain optic and vertical lobe, endothelial cells, pericytes and perivascular glia could be recognized by their morphology and relation to the vascular lumen. In microvessels, endothelial and pericyte membranes showed sparse but uniform distribution of P-face intramembranous particles, with no particular particle aggregations. Perivascular glial membranes had a higher density of intramembranous particles but again no particle alignments characteristic of known restricting junctions were seen, although clusterings of intramembranous particles resembling gap junctions were present. In larger venous vessels, the perivascular glial layer showed a multilamellated organization, but again no arrays of intramembranous particles were detected, although this should be a favourable site for visualization of the restricting junctions. The walls of arterial vessels showed collagen deposits and cell processes with apparent intracellular myofilamentous profiles, but no intramembranous junctional particle arrays. It is concluded that the junctional zone observed in thin section electron microscopy is not associated with aligned aggregations of intramembranous particles detectable in freeze-fracture replicas, strengthening the evidence that this is a novel type of restricting junction.

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PubMed1992

A fibre matrix model for the restricting junction of the blood-brain barrier in a cephalopod mollusc: implications for capillary and epithelial permeability.

A model is proposed for the novel restricting junction forming the blood-brain barrier in a cephalopod mollusc, the cuttlefish Sepia officinalis. The model is based on electron-microscopic findings, from both thin-section and freeze-fracture material, the distribution of electron-dense tracers, and radioisotopic measurements of permeability using small non-electrolytes. Biochemical properties of Sepia plasma proteins are also considered. It is proposed that an effective blood-brain barrier is achieved by a combination of mechanisms. As much as 90% of the Sepia brain microvessel wall is covered by a 'seamless' glial sheath, without intercellular clefts, limiting the number of potential leakage sites. The remaining clefts follow a tortuous course increasing the diffusion path to the neuropile. Entry into the clefts is reduced by a restricting junctional region at the luminal end, characterized by delicate striations spanning the cleft, and forming an effective barrier to both horseradish peroxidase and ionic lanthanum. This is a novel junctional type, different from previously-described vertebrate and invertebrate occluding junctions. It is proposed that the junction acts as a fine-mesh molecular filter, with condensed extracellular material in the cleft, cross-linked and consolidated by bound plasma protein. Cephalopod haemocyanin or its subcomponents are considered likely candidates for the bound protein. The model predicts that blood-brain barrier permeability should be sensitive to the charge structure of the extracellular matrix and the presence of protein, and is analogous to the 'fibre matrix' model of vertebrate capillary permeability. The Sepia blood-brain barrier also highlights the different strategies available for constructing a restricting cell layer, and suggests a possible evolutionary pattern underlying the present range of junctional mechanisms in vertebrate and invertebrate epithelia.

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PubMed1992

Structural domains of the tight junctional intramembrane fibrils.

Freeze-fracture reveals intramembrane fibrils lying along the intermembrane contacts that characterize tight junctions. Tight junctions from a variety of species are reexamined here by rapid freezing prior to freeze-fracture. The tight junction fibril is uprooted alternatively from either the cytoplasmic or the exoplasmic hemibilayer during freeze-cleavage, exposing two distinct but complementary views of its hybrid structure within the same replica. When the transmembrane fibril is uprooted from the exoplasmic hemibilayer it appears on the P-fracture face as a smooth-surfaced cylinder which is sometimes resolved into periodic globular structures. The lack of indication that the P-face cylinder has been pulled out through the opposite membrane half indicates that this domain of the fibril is, in large part, buried in the hydrophobic interior of the membrane. However, when the transmembrane fibril is uprooted from the cytosolic hemibilayer it appears on the E-fracture face as a row of irregular intramembrane particles. The irregular particles on the E-face aspect of the fibril are interpreted as corresponding to transmembrane protein segments that may very well make projections onto the cytosolic surface of the bilayer. En face views of the outermost junction strand between adjacent epithelial cells show periodic lines on the bilayer on each side of the junction which are interpreted as periodic transmembrane protein segments arising from the core structure of the tight junction fibril. If the backbone of the tight junction strand is an inverted cylindrical micelle, it must typically include proteins, which might anchor it to structures outside the membrane bilayer.

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PubMed1991

Morphology of glial blood-brain barriers.

Glial cells, in certain situations in the CNS, may become modified to form the structural basis of the blood-brain barrier. This occurs in more primitive vertebrates, such as the elasmobranch fish, and in some higher invertebrates. In the latter, the outermost glial sheath, often called the perineurium in avascular ganglia, substitutes functionally for the vascular endothelium of higher organisms. The intercellular junctions between the lateral borders of these modified glial or perineurial cells may be of several types. In nearly all cases, adhesive and communicating (gap) junctions are found together with an occluding junctional structure. The latter is assumed to be the morphologic basis of the observed blood-brain barrier. It varies in nature and may be one in which the adjacent cell membranes fuse, partially or completely, to form a classical tight junction, or it may be one in which the cell membranes remain separated by a distinct intercellular cleft. If the latter, the cleft may be straddled by columns or septal ribbons, between which a charged matrix substance may be found. Restrictive linker junctions, recently found to be the basis of the interglial barrier in cephalopod CNS, as well as that of myriapods, are characterized by cross-striations or columns which, in combination with charged residues, inherent either in them or in the associated extracellular matrix, slow down the entry of exogenous molecules. Septate junctions, which occur between glial cells in certain other invertebrates, exhibit intercellular septal ribbons, which do not prohibit paracellular transport of all substances but may slow down the passage of some by virtue of charged moieties. There is an association of cytoskeletal components with these septate, linker, and tight junctions; the role of the cytoskeleton in tight junctions, which can be seen by freeze fracture to be based on simple ridges in insects or a more complex network of them in arachnids, may also be important in the regulation of paracellular permeability. The structural details of the junctions in different groups are summarized and their physiologic implications discussed.

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PubMed1979

Intercellular junctions and the development of the blood-brain barrier in Manduca sexta.

In early embryonic development of the tobacco horn moth no blood-brain barrier is present, as shown by the unimpeded entry of exogenous tracers into the nervous system. However, later on, just before hatching, lanthanum and horseradish peroxidase (HRP) are unable to move inwardly beyond the level of the perineurium, which is the morphological site of the blood--brain barrier in the adult moth, as well as in other insects. Freeze-fracture studies indicate that in the early embryo, 10 nm particles are scattered about in the perineurial membrane PF, either as separate entities or as short linear arrays. By hatching or just before, however, the 10 nm particles have become aligned into lengthy linear aggregates as PF ridges with EF grooves. These would appear to be the simple, arthropod-form of tight junction, and are presumed to be the basis of the perineurial blood-brain barrier. At about the same time, gap junctional elements appear both between adjacent perineurial cells and between glial cells. In both cell types, the gap junctions form from free 13 nm EF particles which gradually become aligned or clumped into strands and aggregates which ultimately coalesce to form first irregular masses and then the macular plaques typical of mature gap junctions. Many of the latter stages are coincident with the hatching of a motile larvae, so that the perineurial and glial cells are by this stage coupled via the channels of the gap junctional particles. They are therefore able to undergo both ionic and metabolic exchange and cooperation during larval life, in addition to being able to respond to hormonal substances in an integrated way. During the 5 larval instars more gap junctions form as the perineurial layer grows thicker. These junctions become more regular in outline and their particles more tightly packed; these larval structures are compared with junctions found in the adult which tend to be more extensive but otherwise similar. Since no septate junctions are apparent during Manduca embryonic or larval life when the blood-brain barrier forms, nor in adults, the results of this report support the contention that it is the tight junctions rather than septate ones which form the basis of permeability barriers in this, and probably other, arthropod systems.

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PubMed1979

Tight junctions in a fluid-transporting epithelium of an insect.

Occluding junctions have been found between the lateral cell borders at the base of the rectum of Periplaneta americana. They appear as punctate membrane appositions in thin sections, and after incubation in physiological solutions containing lanthanum before fixation the inward penetration of tracer is impeded in this same basal area. Moreover, freeze-fracture studies of this region reveal simple linear ridges on fracture face P and grooves on fracture face E, which are similar to the less complex vertebrate tight junctions. The luminal clefts, which permit free inward diffusion of tracers, present no tight junctions, but do have septate junctions. These results support the contention that, contrary to earlier speculation, arthropods do possess tight junctions; these, rather than septate junctions, appear to form the morphological basis of at least some of the permeability barriers observed in invertebrates.

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PubMed1979

Intramembranous particles in the form of ridges, bracelets or assemblies in arthropod tissues.

Non-junctional intramembranous particle arrays in the form of ridges, bracelets or rectilinear assemblies have been found by freeze-fracturing in the cytoplasmic half or P face of the plasma membrane in a variety of arthropod tissues. These tissues include both excitable cells, nerve and muscle, and such other cells as those from the intestinal tract, the tracheal system and the connective tissue. The intramembranous ridges are short rows of fused particles about 10 nm in diameter; comparable particles comprise the bracelets and the rectilinear aggregates, although the former are of lower profile. In cells sending out cytoplasmic projections during migration and development, for example, axons in embryonic, newly hatched or pupal tissues, tracheoles or fibroblasts, the intramembranous ridges are always aligned parallel to the longitudinal axis of the cellular process. The physiological significance of these may be that they play some role in recognition during development, perhaps by contact guidance. The ridges and rectilinear arrays found in the gut could also be involved in recognition and/or adhesion. In muscle, bead-like ridges are intimately associated with the transverse tubular system and may have a receptor function. Irregular and circular low-profile ridges occur in the tissues of the horseshoe crab, Limulus, and 'bracelet' forms are found in the inner membrane of insect pupal tracheae. The latter may play a part in the initiation and development of small tracheoles.

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PubMed1979

Freeze-fracture and tracer studies on the intercellular junctions of insect rectal tissues.

Both rectal pads of the cockroach and rectal papillae of the blowfly possess highly infolded lateral borders; these are associated by desmosomes and septate junctions that maintain the physical integrity of the cell layer at the luminal and basal intercellular regions. Adjacent cells are coupled by gap junctions that allow for cell-to-cell communication and which occur at intervals along the undulating lateral clefts. In rectal pads, occluding basal tight junctions are found as well as extensive scalariform junctions. The latter, like the stacked membrane infoldings of rectal papillae, exhibit intercellular columns and numerous intramembranous P face particles; these are undoubtedly involved in ion transport. In the inter-stack clefts of papillae, reticular septate junctions are encountered which, after freeze-fracture, possess a striking network of PF ridges and EF grooves that are discontinuous and not always complementary. These may serve to regulate the speed and extent of distension of the clefts during solute movement to allow for even and effective fluid flow in this transporting epithelium.

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PubMed1979

An unusual cell surface modification: a double plasma membrane.

The occurrence of an unusual double plasma membrane structure is reported; it has been studied in conventional thin sections, after lanthanum-impregnation and with freeze-fracturing. This modification of the plasmalemma is found where the luminal cell membrane (I membrane) of gut microvilli in the haematophagous insect, Rhodnius prolixus, is surrounded by a second, outer membrane (O membrane), the 2 separated from one another by a highly regular I-O space of about 10 nm. Lanthanum impregnation reveals the presence of columns inclined at an angle, within this I-O space; as in the continuous junctions which link the lateral borders of these cells, these columns may maintain the very precise I-O distance. From the outer microvillar membranes radiate short spoke-like fibrils or sheets which encounter another more extensive system of myelin-like sheets. Freeze-fracturing reveals that the spoke-like sheets and the other ones which lie like a tube, around and parallel to the microvilli, contain linear ridges composed of particles, lying at random within layers of the myelin-like material which also extends into the lumen of the gut. The microvillar membanes, both O and I, fracture into faces containing rows of either PF particles or EF pits arranged as spiral ridges or grooves around the sides and across the tip of each microbillus. These could be the insertion sites of one or both of the I-O columns and spoke-like sheets while the sheets could represent a variant of peritrophic membrane. The double membrane may be a cellular device to increase the strength of the microvillar layer in these blood-sucking animals, since the cell layer must withstand great pressure owing to a sudden massive extension of the gut during a blood meal.

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PubMed1978

Developmental stages in the formation of inverted gap junctions during turnover in the adult horseshoe crab, Limulus.

Stages leading to the formation of inverted gap junctions between certain basal replacement or interstitial cells in the mid-gut of adult Limulus can be followed by freeze-fracturing. Free, 13-nm EF intramembranous particles first appear to be organized into short linear arrays or small clusters of particles, which then become transformed into anastomosing particulate networks covering a considerable surface area. These subsequently become concentrated into smaller, more nearly circular, macular plaques of EF particles or PF pits. These EF particles, both when free or assembled into macular arrays, possess a central channel or pore. Numerous formed gap junctions are present in Limulus mid-gut, which suggests that cell-to-cell communication is an important feature of the mature tissue. The results show that arthropod tissues can be used to study the development of gap junctions not only in differentiating systems but also in adult tissues during normal cell turnover.

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PubMed1977

Lanthanum penetration in crayfish nervous system: observations on intact and 'desheathed' preparations.

Central neural connectives and peripheral nerves from the crayfish Procambarus clarkii were incubated in 5 mM lanthanum solutions in physiological saline, for periods from 15 min to 2 h. The tracer only rarely reaches the axon surfaces in the perineurium-ensheathed connectives, penetrating the elaborate perineurial layer slowly. In peripheral nerves, on the other hand, where the perineurium is extermely attenuated and interrupted by open extracellular clefts, inward movement of lanthanum to the axon surfaces occurs readily. When the perineurial layer of the neural connectives is removed by 'desheathing', penetration of the tracer to the level of the axolemma occurs rapidly, implicating the perineurium as the major site of restriction of entry of large ions and exogenous molecules. This conclusion is discussed in relation to recent electrophysiological studies on K+ movements. In both peripheral nerves and desheathed connectives, the transcellular tubular lattice system present in crustacean glial cells appears to serve as a route for the entry of tracer to the axon surfaces, and is more direct than the long and complex extracellular pathway formed by the interdigitations of the extensive glial cell processes.

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PubMed1977

Intercellular junctions in the central nervous system of insects.

The intercellular junctional complexes in the central nervous system (CNS) from a variety of insect species have been examined by thin-sectioning and freeze-fracturing techniques. Of particular concern has been the fine-structural basis of the blood-brain barrier observed to be present in the outer perineurial layer around the avascular insect CNS. The basis of this has been found in the form of tight junctions (zonulae occludentes) present both in sections and in replicas of the perineurium. In the latter, they appear as one or two simple linear ridges, lying parallel to the outer surface, which occasionally display overlapping. The complex geometry of the interdigitating perineurial cells apparently permits such a relatively simple series of ridges to function as a barrier, since tracers are found not to penetrate beyond this level into the underlying nervous tissue. Such evidence is supported by microprobe X-ray analysis of lanthanum-incubated tissues, the perineurium compared with the glia-ensheathed axons showing the presence and absence of lanthanum, respectively. Possible physiological mechanisms that could operate 'in vitro' to maintain the blood-brain barrier are also considered. Other intercellular junctions such as desmosomes, septate junctions and gap junctions are found in the perineurial layer too, the last exhibiting EF particle plaques and PF pits. Glia-glia junctions also occur in some insect species; they include desmosomes, inverted gap junctions and occasional tight junctions. Septate, gap and tight junctions are also found on the membranes of tracheoles penetrating the CNS. Short, ridge-like elaborations and other particle arrays are found on the PF on the axon surfaces and the significance of these structures is discussed.

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