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N J Lane

Publications and source records attributed to N J Lane.

At least 73 records · Page 4Linked to original sources
PubMed1980

Definitive evidence for the existence of tight junctions in invertebrates.

Extensive and unequivocal tight junctions are here reported between the lateral borders of the cellular layer that circumscribes the arachnid (spider) central nervous system. This account details the features of these structures, which form a beltlike reticulum that is more complex than the simple linear tight junctions hitherto found in invertebrate tissues and which bear many of the characteristics of vertebrate zonulae occludentes. We also provide evidence that these junctions form the basis of a permeability barrier to exogenous compounds. In thin sections, the tight junctions are identifiable as punctate points of membrane apposition; they are seen to exclude the stain and appear as election- lucent moniliform strands along the lines of membrane fusion in en face views of uranyl-calcium-treated tissues. In freeze-fracture replicas, the regions of close membrane apposition exhibit P-face (PF) ridges and complementary E-face (EF) furrows that are coincident across face transitions, although slightly offset with respect to one another. The free inward diffusion of both ionic and colloidal lanthanum is inhibited by these punctate tight junctions so that they appear to form the basis of a circumferential blood-brain barrier. These results support the contention that tight junctions exist in the tissues of the invertebrata in spite of earlier suggestions that (a) they are unique to vertebrates and (b) septate junctions are the equivalent invertebrate occluding structure. The component tight junctional 8- to 10-nm-particulate PF ridges are intimately intercalated with, but clearly distinct from, inverted gap junctions possessing the 13-nm EF particles typical of arthropods. Hence, no confusion can occur as to which particles belong to each of the two junctional types, as commonly happens with vertebrate tissues, especially in the analysis of developing junctions. Indeed, their coexistance in this way supports the idea, over which there has been some controversy, that the intramembrane particles making up these two junctional types must be quite distinct entities rather than products of a common precursor.

Animals↗
PubMed1980

Electrical interactions between the giant axons of a polychaete worm (Sabella penicillus L.).

Intracellular recordings demonstrated a transfer of impulses between the paired giant axons of Sabella, apparently along narrow axonal processes contained within the paired commissures which link the nerve cords in each segment of the body. This transfer appears not to be achieved by chemical transmission, as has been previously supposed. This is indicated by the spread of depolarizing and hyperpolarizing voltage changes between the giant axons, the lack of effects of changes in the concentrations of external divalent cations on impulse transmission and by the effects of hyperpolarization in reducing the amplitude of the depolarizing potential which precedes the action potentials in the follower axon. The ten-to-one attenuation of electronic potentials between the giant axons argues against the possibility of an exclusively passive spread of potential along the axonal processes which link the axons. Observation of impulse traffic within the nerve cord commissures indicates, on the other hand, that transmission is achieved by conduction of action potentials along the axonal processes which link the giant axons. At least four pairs of intact commissures are necessary for inter-axonal transmission, the overall density of current injected at multiple sites on the follower axon being, it is presumed, sufficient to overcome the reduction in safety factor imposed by the geometry of the system in the region where axonal processes join the giant axons. The segmental transmission between the giant axons ensures effective synchronization of impulse traffic initiated in any region of the body and, thus, co-ordination of muscular contraction, during rapid withdrawal responses of the worm.

Action Potentials↗
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.

Animals↗
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.

Animals↗
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.

Animals↗
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.

Animals↗
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.

Animals↗
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.

Animals↗
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.

Animals↗
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.

Animals↗
PubMed1976

Interrelationships between Golgi, GERL and synaptic vesicles in the nerve cells of insect and gastropod ganglia.

In addition to demonstrating synaptic vesicles, staining with the zinc-iodide-osmium tetroxide (ZIO) method reveals the presence of positively reacting GERL membranes in association with the Golgi complex and lysosomes in the nerve cell bodies within ganglia from the locust Schistocerca gregaria and the gastropod molluscs, Limnaea stagnalis and Helix aspersa. A positive response to ZIO occurs in certain Golgi vesicles and saccules, in GERL (Golgi-endoplasmic-reticulum-lysosomes), in multivesicular bodies as well as residual bodies and in small vesicles and cisternae of axonal smooth endoplasmic reticllum (ER). The interrelationships between these organelles are considered in view of the similarity of the ZIO localization to phosphatase-rich sites in the neuronal perikarya and with respect to the possibility that components of the synaptic vesicles are formed in the Golgi region of the cell and migrate via the axonal smooth ER to the synaptic regions.

Animals↗
PubMed1975

The organization of the nervous system in the crayfish Procambarus clarkii, with emphasis on the blood-brain interface.

Central neural connectives and peripheral nerves of the crayfish Procambarus clarkii are surrounded by an acellular neural lamella, beneath which lies a layer of specialised glia, the perineurium. Cell process of the connective perineurium interdigitate extensively, and are frequently closely associated with each other by gap junctions. Occasional zonulae occludentes are encountered. Nerve perineurium, however, is much less elaborate, and may be reduced to a single or incomplete cell layer. In both connective and nerve, the perineurium appears involved in the formation of the collagen-like fibrils of the neural lamella. The comparative fine structure of connective and peripheral nerve correlates well with recent experimental studies in crayfish, where it was concluded that the perineurium in connective but not nerve offers some restriction to diffusion of small ions and molecules. Within the connective, deeper glia re either closely associated with axons (Schwann cells) or lie relatively free in the extracellular space. Cytoplasmic process of both cell types possess "tubular lattice" systems, which are especially elaborate in the Schwann cells. The extracellular space contains a flocculent material and bundles of collagen, together with layers of basal lamina-like material. The physiological implications of the observations are discussed.

Animals↗
PubMed1975

Insect peripheral nerves: accessibility of neurohaemal regions to lanthanum.

During incubation in vivo, exogenously applied ionic lanthanum comes to surround the numerous neurosecretory terminals which are found lying within or immediately beneath the acellular neural lamella ensheathing the nerves from fifth instar and adult specimens of Rhodnius prolixus. The lanthanum does not penetrate beyond the cellular perineurium, which completely surrounds the non-neurosecretory axons in these nerves and constitutes a form of 'blood-brain barrier'. In some cases, however, lanthanum is found in the vicinity of a neurosecretory axon lying beneath the perineurium, where it can be assumed to have leaked in from the neurosecretory terminal lying free in the neural lamella. When nerves are incubated in calcium-free media, regions with an attenuated perineurium become 'leaky', in that lanthanum is found lying in those extracellular spaces between axons and glia which lie immediately below the thin part of the perineurial layer. Bathing solutions made slightly hyperosmotic to the haemolymph with sucrose have no apparent disruptive effects on the barrier. When the tissues are incubated in more hypertonic solutions, the perineurial barrier becomes 'leaky' throughout, and tracer pervades beyond its cells into all the intercellular spaced between glia and axons. The possible role of the zonulae occludentes in both the maintenance of the perineurial barrier and in the formation of interglial occlusions to local penetration of exogenous substances is considered.

Animals↗