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N J Lane

Publications and source records attributed to N J Lane.

At least 55 records · Page 3Linked to original sources
PubMed1984

Isolation and characterisation of arthropod gap junctions.

Gap junctions have been isolated from the hepatopancreas of the crustacean arthropod, Nephrops norvegicus (Norway lobster). SDS-PAGE of these preparations shows two major protein bands, mol. wt. 18 000 (18 K) and mol. wt. 28 000 (28 K). The 18-K and 28-K proteins are interconvertible, cannot be distinguished by two dimensional tryptic and chymotryptic peptide mapping, and therefore appear to be different (most likely monomeric and dimeric) forms of the same protein. The protein can also aggregate to higher multimeric forms mol. wt. 38 000 (presumed trimer), and mol. wt. 52 000 (presumed tetramer). The buoyant density of the isolated gap junctions in continuous potassium iodide gradients is 1.260 g/cm. The junctions are progressively solubilized in increasing SDS concentrations, mostly between 0.1% and 0.2% SDS, and this is accompanied by the release of the 18-K and 28-K forms of the junctional protein. The Nephrops hepatopancreas 18-K junctional protein has antigenic determinants in common with the vertebrate 16-K junctional protein as shown by cross-reactivity with two different affinity purified antibody preparations. However, no detectable similarity can be seen between the major I-labelled tryptic and chymotrytpic peptides of the Nephrops hepatopancreas 18-K protein and the mouse liver 16-K protein.

Journal Article↗
PubMed1984

Intercellular junctions in the hepatopancreas of the lobster Nephrops norvegicus.

The hepatopancreas of the lobster has recently been found to be a rich source of material from which to isolate arthopod gap junctions biochemically (Finbow et al., 1983a; 1984). It has therefore been studied here to assess the features of these intercellular junctions and any others that may be present, in vivo. The tissue consists of columnar epithelial cells which possess apical microvilli and basal infoldings. In thin sections the lateral borders of these cells are characterized by desmosomes and smooth septate junctions as well as by gap junctions. The desmosomes exhibit no apparent freeze fracture profile but the septate junctions display parallel rows of ridges or aligned intramembranous particles (IMPs) with complementary grooves on the other membrane half; these IMPs shift in their preferential fracturing plane depending on whether the tissue has first been fixed, always remaining on the EF if unfixed. The IMPs or connexons, of which the gap junctions are composed, fracture onto the E face, leaving complementary pits on the P face, regardless of whether the tissue is fixed or not. At the base of the pancreatic cells, the lateral borders are thrown into interdigitating folds which display endocytotic profiles and possible internalization of junction-bearing membranes. This phenomenon, which is readily visualized both after tracer incubation and in replicas, may represent junctional degradation relating to membrane turnover.

Animals↗
PubMed1984

Glial repair in an insect central nervous system: effects of surgical lesioning.

Surgical lesioning of central nervous connectives in the cockroach (Periplaneta americana (L.], although causing only local glial damage, resulted in complex and prolonged cellular changes. An early response to mechanical disruption was the appearance of granule-containing cells within the damaged perineurium, among adjacent, undamaged, perineurial cells, and between glial processes deep within the connectives. These cells, which were strikingly similar to hemocytes, were clearly involved in phagocytic activity and persisted in the damaged regions for more than a month after lesioning. There was only a slow restoration of organized perineurial glia and re-establishment of the blood-brain barrier, as indicated by the exclusion of an extracellular tracer, ionic lanthanum. These observations contrast with the speedy, ordered repair of the neuroglia observed following selective glial disruption and suggest that undamaged axons and/or the extracellular matrix exert a profound influence on the mechanisms of glial repair.

Animals↗
PubMed1983

Myelinating glia of earthworm giant axons: thermally induced intramembranous changes.

The median and lateral giant axons in the ventral nerve cord of the earthworm Lumbricus terrestris are ensheathed by extensive spiral glial cell wrappings which resemble vertebrate myelin. The other, smaller, axons are encompassed by attenuated glial processes, as is typical of invertebrates. The fine structural details of the glial cells have been studied in thin sections and in replicas produced by freeze-fracturing where the intramembranous particle (IMP) populations within the lipid bilayer are visible. These consist of both low-profile IMPs as well as prominent ones 6-8 nm in diameter, scattered at random over the lipid interface in the myelinating glia. The larger IMPs on both P and E faces number about 80/mum2 at 16 degrees C in contrast to the IMP density of 400/mum2 in the other glial membranes. After acclimation to 5, 16 and 26 degrees C, the loose myelin glial membranes show variations in the density of their larger IMP population; in animals acclimated over 3 or more weeks to 5 degrees C, the number of these IMPs is significantly (P less than 0.001) less per unit area than in animals acclimated to 16 or 26 degrees C. The size of the particles at 5 degrees C is significantly (P less than 0.001) smaller than those at 16 or 26 degrees C. When animals are subjected to a sudden differential in ambient temperature, from 26 or 16 to 5 degrees C, or from 5 to 26 degrees C, and their giant axons with encompassing glia are fixed and frozen 30 min after this temperature change, the IMP population of the glial membranes remaining does not appear to alter. The differences in the IMP population of the myelinating glial membranes at different temperatures may reflect the extent to which they insulate and/or influence the velocity of impulse propagation.

Acclimatization↗
PubMed1983

Pre- and post-synaptic structures in insect CNS: intramembranous features and sites of alpha-bungarotoxin binding.

The central neuropile of thoracic ganglia in the central nervous system (CNS) of the cockroach Periplaneta americana contains synapses with characteristic pre- and post-synaptic membrane specializations and associated structures. These include dense pre-synaptic T-bars surrounded by synaptic vesicles, together with post-synaptic densities of varying electron opacity. Exocytotic release of synaptic vesicles is observed only rarely near presynaptic densities, but coated pits are seen at variable distances from them, and may be involved in membrane retrieval. After freeze-fracture, paralinear arrays of intramembranous articles (IMPs) are detected on the P face of many presynaptic terminals, with associated dimples indicative of vesicular release. The E face of these membranes exhibits protuberances complementary to the P face dimples, as well as scattered larger IMPs. Post-synaptic membranes possess dense IMP aggregates on the P face, some of which may represent receptor molecules. Electrophysiological studies with biotinylated alpha-bungarotoxin reveal that biotinylation does not inhibit the pharmacological effectiveness of the toxin in blocking acetylcholine receptors on an identified motoneurone in the metathoracic ganglion. Preliminary thin section ultrastructural analysis of this tissue post-treated with avidin-HRP or avidin-ferritin indicates that alpha-bungarotoxin-binding sites are localized at certain synapses in these insect thoracic ganglia.

Animals↗
PubMed1983

Insect intercellular junctions: rapid freezing by jet propane.

The ventral nerve cord of the cockroach, Periplaneta americana, and that of the locust, Schistocerca gregaria, have been studied after rapid freezing by cryo-jet, using liquid propane. Such tissues, unfixed and uncryoprotected, have been compared with unfixed cryoprotected tissues, as well as with material fixed with glutaraldehyde and cryoprotected with glycerol or polyvinylpyrrolydone. The perineurial tight junctions in the cryo-jet-frozen tissues exhibit characteristic intramembranous P-face ridges, but frequently these are composed of smooth-surfaced strands, comparable to those seen with fast-freezing in mammalian tissues, rather than of bead-like fibres. The intramembranous PF ridges, characteristic of axonal and glial processes in the insect central nervous system, also display a smooth surface in rapidly frozen preparations. Prior fixation and/or cryoprotection produces a bead-like appearance in the ridges. The interglial gap junctions, after fast-freezing, exhibit both clustered connexon arrays in the E-face and loosely aggregated ones; hence the coupled state cannot be unequivocally associated with the latter configuration. The septate junctions between glial cells are unchanged after rapid freezing, exhibiting the typical rows of P-face intramembranous particles with complementary E-face pits that are found in replicas from fixed and cryoprotected tissues. The surfaces of the axons and glial processes exhibit pleiomorphic depressions and associated particles as well as PF pits with complementary EF mounds, both with associated IMPs. These structures are not usually seen after fixation or cryoprotection and may represent some kind of receptor structure, or axo-glial specialization.

Animals↗
PubMed1982

Physiological and ultrastructural evidence for an extracellular anion matrix in the central nervous system of an insect (Periplaneta americana).

The efflux of radiocations (22Na, 2K and 45Ca) and of radiochloride occur as two-stage processes from intact cockroach nerve cords. It is suggested that the initial, fast fraction of efflux comes mainly from the superficial connective tissue layer, the neural lamella, and the clefts between the underlying layer of neuroglia, the perineurium. This is deduced from the lack of effect of a metabolic inhibitor and sodium-transport inhibitors on the fast component of 22Na efflux (which contrast with their effects both on the size an the half-time of the slow component) and from the typically extracellular ratios between the fast components of substantial increase in the fast fractions of 22Na and 45Ca efflux but only a small increase in 36Cl efflux: effects which would be expected if the addition to the fast fraction consisted of ions maintained in Donnan equilibrium with fixed anionic sites within the extracellular system. The presence of such anionic sites is also indicated by lanthanum-binding in the extracellular matrix and by the previous histochemical demonstration of hyaluronic acid in the matrix by Ashhurst and Costin. It is suggested that the anionic glycosaminoglycans provide an extracellular cation reservoir which could serve a role in short-term ionic homeostasis of the brain microenvironment.

Animals↗
PubMed1982

Presence of T-bars, intramembranous particle arrays and exocytotic profiles in neuroendocrine terminals of an insect.

Definitive evidence is presented to show that arthropod neurohaemal terminals contain electron-dense T-bar structures with clustered microvesicles similar to those present in neuropilar and neuromuscular terminals. In terminal membranes of the locus corpus cardiacum, studied by freeze-fracture, intramembranous particle arrays, considered to correlate with the dense bars, are seen. However, there does not appear to be a spatial association between the arrays and the exocytotic profiles seen following exposure to stimulants for hormone release. The presence of the densities in both neuroendocrine and conventional terminals is discussed in the light of current theories for mechanisms of release of neurotransmitters and neurohormones in arthropods and vertebrates.

Animals↗
PubMed1982

Differential accessibility to two insect neurones does not account for differences in sensitivity to alpha-bungarotoxin.

The nicotinic acetylcholine receptor probe alpha-bungarotoxin (1.0 x 10(-7) M) blocks the depolarising response to ionophoretic application of acetylcholine onto the cell body membrane of the fast coxal depressor motoneurone (Df) of desheathed cockroach (Periplaneta americana) metathoracic ganglia, but at the same concentration is completely ineffective in blocking the depolarising action af acetylcholine on dorsal unpaired median (DUM) neurones in the same ganglion. The possibility that this is due to differences in accessibility of the toxin to the neurones has been tested by a combination of ionophoretic injection of horseradish peroxidase into single neurones with a study of the distribution of the exogenous tracer lanthanum, which is of similar effective size to alpha-bungarotoxin. The peripherally located cell body membranes and the fine axonal processes of Df and DUM neurones of desheathed metathoracic ganglia are equally accessible to lanthanum. Differential accessibility to the two cell types does not account therefore for the differences in sensitivity to alpha-bungarotoxin.

Acetylcholine↗
PubMed1982

Stages in the assembly of pleated and smooth septate junctions in developing insect embryos.

The stages that occur during the assembly of both pleated and smooth septate junctions in developing insect tissues have been examined. The oesophagus and mid-gut of the embryonic moth, and the oesophagus and central nervous system (CNS) of the locust embryo, have been investigated in thin sections and by freeze-fracture during the course of membrane biogenesis. The smooth septate junctions developing between the lateral borders of the mid-gut exhibit, in the early stages, individual intramembranous particles becoming aligned into short ridges. These ultimately migrate over the membrane face and fuse into longer arrays, which become stacked in parallel with other ridges to form the characteristic mature form of the junction just before hatching. Pleated septate junctions occur between the cells both of the oesophagus and of the perineurium, which ensheathes the neurones and the neuroglial cells in the locust CNS; these are also fully formed by the end of embryonic development. The pleated junctions appear to be assembled during the later stages of CNS or gut differentiation, arising first in embryos about two-thirds of the way through development. During their maturation, the initial event seems to be a membrane depression in the P face, which occurs in patches over the presumptive junctional membrane. Into these depressed regions or 'formation-plaque' areas, 8-10 nm particles appear to be inserted intramembranously in apparently random arrays. These particles are the most common elements but larger particles are also present; the former ultimately become aligned in a row. With time, other intramembranous particles come to lie in rows parallel to the original one. By hatching, the typical undulating stacks of parallel intramembranous particle rows are fully formed. Gap junctions also form between the same perineurial or oesophageal cells, usually before, but in some cases at the same time, or just after, the septate junctions have been assembled. Tricellular associations between cells also appear around the same time in embryonic development. The simultaneous assembly of these different junctions reflects a high degree of organizational capacity at the membrane level.

Animals↗
PubMed1982

Carbon dioxide induced disassembly of gap-junctional plaques.

When sheep ovarian follicle cells are maintained in an O2-rich environment their cells are metabolically coupled, as monitored by observing the exchange of [3H]choline; choline metabolites were detected up to 4 mm from the explant under these control conditions. When the tissues were placed in a CO2-rich environment the cells became uncoupled physiologically and choline metabolites were no longer exchanged. The cells in these two states, coupled and uncoupled, were examined by freeze-fracture. The initial controls were characteristic of ovarian follicular tissue exhibiting large macular plaques with regular outlines composed of PF intra-membranous particles (IMPs), which were arrayed in rows with IMP-free aisles. With uncoupling, the junctional plaques became irregular at the periphery, they became loosely packed and IMPs began to 'stream' out laterally across the membrane. Ultimately they were reduced to negligible IMP clusters or free IMPs. Analyses of the IMPs with an image analyser confirmed that in the uncoupled state the gap-junctional IMPs were dispersed over the membranes. On return to an O2-rich environment, the cells became recoupled as monitored by physiological criteria and in freeze-fracture replicas IMPs reclustered into macular, albeit smaller, plaques. These results support the contention that with uncoupling, gap-junctional particles are free to move and hence may become dispersed over the membrane face, with the possibility of being re-utilized to form junctions anew when conditions for coupling are re-established.

Animals↗
PubMed1981

Lack of restriction at the blood-brain interface in Limulus despite atypical junctional arrangements.

Tracer and freeze-fracture techniques are used to evaluate the capacity of the central and peripheral nervous system of the horseshoe crab, Limulus polyphemus to admit or exclude molecular or ionic constituents of the blood intercellularly. Both the peripheral and central nervous systems are contained within blood sinuses so there is intimate contact between the haemolymph and the neural lamella. No discrete perineurium exists so any protection afforded to the nerve cells must be provided by the ensheathing glial cells and any junctions between them. Using ionic lanthanum as a pre-fixation incubation medium the system is seen to be completely "open', with the tracer gaining access to all regions of the nervous tissue. Cellular association in the peripheral nervous system, as revealed by thin-section and freeze-fracture, consist only of small scattered gap junctions between glial cells which afford no restriction to tracer entry. Gap junctions are again present between glial cells in the C.N.S. but here they are far more numerous, sometimes forming extensive sheets of almost continuous gap junctional plaques. Between certain glial cells there also exists a junctional system of linear PF ridges and complementary EF grooves; these may associate with or surround, often discontinuous arrays, the gap junctional plaques. Given their characteristics and the freedom of tracer entry, they seem unlikely to represent either typical occluding tight junctions or septate junctions.

Animals↗
PubMed1981

A vertebrate-like blood--brain barrier, with intraganglionic blood channels and occluding junctions, in the scorpion.

India ink and ionic lanthanum injections have revealed that the central nervous system (CNS) of the scorpion possesses a highly vascularized cephalothoracic ganglionic mass. It, together with other abdominal ganglia which form a ventral nerve cord, are all ensheathed by an outer layer of modified glial, or perineurial, cells. These cells resemble those which line the blood channels permeating the CNS, in exhibiting both inverted gap and tight junctions. Although the latter show close or fused membrane appositions, lanthanum appears to penetrate past a number, but not all, of them. Freeze-fracturing reveals that these junctions are composed of E-face particles aligned into a network of rows, or ridges, which are frequently discontinuous, especially near the periphery of the perineurium. This produces a somewhat 'leaky' system but occlusion to tracers occurs ultimately, for in the CNS none can be found beyond the perineurium. The existence of this perineurial blood-brain barrier is also demonstrable electrophysiologically where cations such as Mg2+ are unable to penetrate beyond the perineurial layer although they can, it seems, leak in via the blood vascular system. Relative differences in tightness between the perineurium and the cells lining the blood channels may be attributed to differences in the relative number of discontinuous ridges. This is borne out by the observation that the peripheral nervous system has a highly attenuated perineurium with many fewer junctions, and some of these nerves tend to be leaky with respect to tracer penetration. In fixed material the junctional ridges may fracture on to the E-face or partly on both the EF and PF, while in unfixed tissue they are usually found on the PF. In both cases they exhibit complementary grooves that are coincident with the ridges across membrane transitions; in such cases the cell membranes are fused with concomitant obliteration of the intercellular space. These tight junctions, often closely associated with EF gap junctional particle aggregates which may be very loosely clustered, appear to form the basis of the observed blood-brain barrier in the scorpion CNS.

Animals↗
PubMed1980

Fusion of liposome membranes by the n-alkyl bromides.

It has been found that the n-alkyl bromides are capable of inducing the fusion of unilamellar liposomes. These compounds can bring about fusion of liposomes composed of either pure phosphatidylcholine or phosphatidycholine+phosphatidic acid. Fusion of unilamellar liposomes gives rise to multilamellar structures, the morphology of which has been examined by negative staining and freeze-fracture techniques. It has been shown by microelectrophoresis that the n-alkyl bromides have no effect on the surface charge of liposomes, and fusion has been further characterized by use of light scattering and differential scanning calorimetry, the latter indicating that true mixing of the fatty acyl chains occurs upon fusion. Finally, fusion occurs at n-alkyl bromide levels below that required to saturate the aqueous phase of the system.

Calorimetry, Differential Scanning↗
PubMed1980

Perineurial and glial cells in the tick Boophilus microplus (Acarina: Ixodidae): freeze-fracture and tracer studies.

In the cattle tick Boophilus microplus, the cells of the perineurium are characterized by accumulations of glycogen which increase dramatically after feeding. Gap junctions couple both these perineurial cells which enshealth the C.N.S. and the underlying glial cells. No tight junctions have been found between perineurial cells and there is in consequence no blood-brain barrier. Using ionic lanthanum as a tracer the extensive gap junctions are shown to have no occluding effect and lanthanum penetrates through the perineurium and glial layers to the level of the axonal surfaces. By colloidal lanthanum impregnation and freeze-fracture studied, the gap junctions appear to be typical of arthropids in that their particles show a characteristic diameter (13 nm in freeze-fracture), are distributed relatively loosely within the junctional plaques and fracture onto the E face of the junctional membranes. Semi-ordered particle arrays are found on E face membranes of adjacent axons and glia which may represent axoglial junctions.

Animals↗