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Effects of horizontal cell network architecture on signal spread in the turtle outer retina. Experiments and simulations.

In the Pseudemys turtle retina five functionally distinct, electrically coupled networks of horizontal cells distribute signals in the outer plexiform layer. These networks differ significantly in their architecture, as determined by intracellular labeling with Neurobiotin after physiological recording and identification. The density of H1 horizontal cells is highest, ranging around 1800 cells/mm2 at approximately 2.3 mm eccentricity. H1 horizontal cell somata are connected via 6-10 thin, short dendrites. The H1 horizontal cell axon terminal network is composed of thick axon terminals, forming a three-dimensional, sheath-like structure. Networks of coupled H2 and H3 horizontal cells have cell densities of around 210 cells/mm2 and 350 cells/mm2, respectively, at the same eccentricity of 2.3 mm. Cell bodies are connected with 6-12 long, thin dendrites. Here we report for the first time H4 horizontal cell networks. Cell density is approximately 970 cells/mm2 at 2 mm eccentricity, and cell bodies are connected with 6-10 thin, short dendrites. General properties of passive voltage spread were compared for three of these horizontal cell networks using NeuronC. Realistic network architectures were obtained by digitizing the intracellularly labeled networks, respectively. One network obtained from coupled H1 horizontal cell bodies, one from coupled H1 horizontal cell axon terminals, and one from H2 horizontal cells were simulated. These three realistic networks were compared with an artificial, electrically coupled regular triangular network. Passive signal spread in these networks strongly depended on the exact network architecture using otherwise identical parameters. Changes in coupling strength affected signal spread in these networks differently. As in the experimental situation, changes in synaptic conductance influenced signal spread. Some principal effects of extensively coupled horizontal cells on photoreceptor signal processing were simulated with one type of photoreceptor connected by telodendria, synapsing onto an underlying triangular network and receiving feedback synapses. Under certain conditions, spatial information is coded in single photoreceptors. This was also the case in the experimental situation. In the simulation, spatial filter adjustment for optimal spatial coding in photoreceptors can be achieved by changing coupling strength in the horizontal cell network.

Animals↗

Spread of plant virus disease to new plantings: a case study of rice tungro disease.

The spread of plant virus disease between fields is reviewed for several horticultural and agricultural crops. Previous studies have focused on cropping systems where there is scope for using separation in time and space to reduce the potential for virus spread between plantings. In our study, data are presented on virus disease spread between fields in an irrigated rice area in the tropics where crops are grown continuously throughout the year. An intensive survey was conducted in rice fields planted from November 1992 to October 1994 in which the occurrence of new infections of rice tungro virus disease was recorded. The effect of a number of variables on disease incidence, including the proximity of a field to inoculum sources, was examined using ordinal regression analysis. Primary infection showed large seasonal fluctuation. In addition, the number of leafhopper vectors had a significant effect, as did the tungro vector resistance of the rice variety grown. The distance to nearest inoculum source and the tungro incidence of this source significantly affected the level of infection occurring in a vulnerable field. The results are discussed in relation to management strategies to reduce the spread of tungro disease between fields in irrigated lowland rice cropping systems.

Crops, Agricultural↗

Periodontal ligament injection: spread of the solution in the dog.

Investigations of the spread of anesthetic solutions administered with the periodontal ligament injection have reported conflicting findings. The purpose of this investigation was to study the distribution of the injected fluid by macroscopic and microscopic techniques. One third of the liquid of the carpules (2% lidocaine with 1:100,000 epinephrine) was replaced with India ink as a marker. Injections were made with a pressure syringe, (Ligmaject) at 90 sites in 12 dogs. Twenty-five control sites were injected with a marked solution without a vasoconstrictor (3% mepivacaine). Seven dogs were killed within 24 h; the others after 5 days. Spread of the ink was studied in undemineralized slices, in three-dimensional cleared specimens, and in histological sections. The solution usually reached the alveolar crest, seeped under the periosteum, and entered into bone marrow spaces alongside vascular channels. The ink spread along the path of least resistance, influenced by the intricacies of anatomical structures and fascial planes, rather than penetrating into the periodontal ligament or in the root canal. In the controls without a vasoconstrictor the spread was more diffuse. Five days postoperatively, carbon particles were present only within macrophages.

Anesthesia, Dental↗

Is migraine explained by Leao's spreading depression?

Spreading depression (SD) is a severe but transient disruption of neural activity in the brain, which spreads like waves in a pond in which a stone has been cast. It propagates to normal tissues and is accompanied by flux of Na+, Ca++, and Cl- ions into the cells, resulting in a brief burst of action potentials followed by electrical silence. Its rate of spread correlates almost exactly with the observed spread of the aura of classical migraine. It is accompanied in animal experiments by changes in regional cerebral blood flow which closely simulate changes observed in man during the migraine aura. There are serious difficulties in extrapolating from SD in animals to migraine in man, but the possibility of SD as the initiating event accords with the notion of migraine as a cerebral (neural) disorder and encourages further investigation in man.

Brain↗

Movement of fibrinogen receptors on the ventral membrane of spreading platelets.

We have used fibrinogen gold conjugates and fluorescein-labelled fibrinogen to examine the presence of fibrinogen receptors on the ventral membrane of spreading platelets and to visualize the redistribution of the receptors during platelet spreading. The movement of fibrinogen gold conjugates was observed in real time using video-enhanced interference reflection microscopy (VEIRM) and the redistribution of fluorescein-labelled fibrinogen was examined using video-intensified epifluorescence microscopy (VIFM). Fibrinogen was picked up by spreading platelets and moved towards the centre of the platelets. The velocity of the movement of fibrinogen-gold conjugates under the ventral membrane ranged from less than 0.1 microm min(-1) to more than 10 microm min(-1). The average velocity in the peripheral web region was more than 6 microm min(-1), and was reduced to around 1 microm min(-1) in the outer and inner filamentous zones. The centralized fibrinogen molecules were internalized and eventually removed from the platelets. Video microscopy in conjunction with colloidal gold particles provides a simple method for real-time observation of the movement of fibrinogen receptors on the ventral membrane of spreading platelets.

Adsorption↗

Microdistribution of substratum-bound ligands affects cell function: hepatocyte spreading on PEO-tethered galactose.

Cell functions are regulated by signal transduction processes triggered by binding interactions with extracellular ligands, including those coupled to insoluble matrices as well as those diffusing in free solution. Whereas receptor interactions with freely diffusible soluble ligands are quantitatively governed by the mean ligand concentration, interactions with spatially constrained substratum-bound ligands may be affected not only by the mean ligand concentration, but also by the ligand spatial microdistribution. To probe this hypothesis we have generated surfaces presenting galactose ligands at different concentrations and different capabilities for micromobility--and thus spatial microdistribution--by means of polyethylene oxide tethering and assayed for hepatocyte spreading mediated by asialoglycoprotein receptor/cytoskeleton linkages. We demonstrate that spreading is not uniquely determined by the mean galactose concentration presented by the culture substrate. Rather, the ability of primary hepatocytes to spread is additionally specified by a combination of ligand concentration and tether length. Our results suggest spreading results when monovalent ligands possess sufficient mobility to form effectively multivalent bonds with the receptor; that is, when the substratum-bound ligands to cluster in spatial microdomains.

Animals↗

Molecular mechanisms of neurotropic herpesvirus invasion and spread in the CNS.

Pseudorabies virus (PRV) is a herpesvirus in the subfamily alphaherpesvirinae (the alpha herpesviruses). After primary infection at mucosal surfaces, PRV infects the peripheral nervous system in its natural host (swine) with occasional invasion of the central nervous system. When other hosts (including cows and rodents) are infected, the infection almost always gives rise to fatal disease in the CNS as a result of infection of peripheral neurons and subsequent spread to the brain. Part of the ability to cause fatal CNS disease can be attributed to a viral glycoprotein called gE. Viruses lacking gE are thought to be less virulent because they do not spread efficiently from cell to cell. Based on a set of gE mutations we have constructed, we suggest that these two phenotypes of cell-cell spread and virulence reflect separate functions of the gE protein. In this report, we show that viruses carrying these new gE mutations have marked reduction in virulence, yet spread efficiently in defined neural circuits in the rat brain. As such, they offer new insight and opportunities for understanding of viral disease and host response to injury, as well as in the construction of viral tracers of neuronal connections.

Animals↗

Time course and effective spread of lidocaine and tetrodotoxin delivered via microdialysis: an electrophysiological study in cerebral cortex.

Microdialysis is a useful tool for administering drugs into localized regions of brain tissue, but the diffusion of drugs from the probe has not been systematically examined. Lidocaine (10%) and tetrodotoxin (TTX, 10 microM), drugs typically used in neural inactivation studies, were infused through a microdialysis probe into raccoon somatosensory cortex while evoked responses were recorded at four electrodes equally spaced 0.5--2.0 mm from the probe. The decreases in evoked response amplitude as a function of time and distance from the probe were used as functional measures to describe the time course and spread of the drugs. TTX inactivated distant sites more quickly and to a greater extent than lidocaine. Responses recovered within approximately 40 min after termination of lidocaine, but did not recover for at least 2 h after TTX. Based on these measurements, we estimated that, at the concentrations used, lidocaine has a maximal spread of 2.1 mm, while TTX could spread as far as 4.8 mm from the microdialysis probe. However, in terms of significant inactivation of neuronal activity, lidocaine and TTX have an effective spread of 1 and 2 mm, respectively.

Anesthetics, Local↗

Beta 1 integrins regulate axon outgrowth and glial cell spreading on a glial-derived extracellular matrix during development and regeneration.

In the present study we have investigated functional roles for beta 1 integrin receptors in regulating axon outgrowth, and glial cell adhesion and spreading in the Xenopus retina. The XR1 glial cell line, isolated from Xenopus retinal neuroepithelium, deposits a proteinaceous extracellular matrix (ECM) with potent neurite outgrowth promoting activity. To investigate a potential role of the integrins as cellular receptors for these glial cell-derived ECM components, embryonic and regenerating retinal explants were cultured in the presence of polyclonal antibodies directed against the beta 1 integrin receptor complex. The IgGs and Fabs of the anti-beta 1 integrin antibody strongly inhibited ganglion cell axon outgrowth on the glial cell-derived ECM, although axons grew freely across the surfaces of glial cells surrounding the explants. The antibodies also inhibited outgrowth on purified laminin containing substrates in a dose-dependent fashion. In addition, the anti-beta 1 antibodies were effective at inhibiting the spreading of glial cells that migrated out from the embryonic explants, and also inhibited attachment and spreading of Xenopus XR1 glial cells on ECM substrates. These results show that the beta 1 integrins play important functional roles in axon outgrowth during development and regeneration, and also serve in regulating retinal glial cell attachment and spreading in vitro, and thus are likely to play similar roles in vivo.

Animals↗

Integrin alpha IIb beta 3-mediated pp125FAK phosphorylation and platelet spreading on fibrinogen are regulated by PI 3-kinase.

Activation of the focal adhesion kinase pp125FAK correlates with its phosphorylation on tyrosine residues and is mediated by multiple receptor-ligand pairs. In platelets, pp125FAK phosphorylation is triggered by alpha IIb beta 3 integrin or Fc gamma RII receptor interaction with immobilized fibrinogen and IgG, respectively. In this study we used platelets as a model system to explore the role of PI 3-kinase relative to pp125FAK phosphorylation. Treatment of the platelets with two PI 3-kinase inhibitors, wortmannin and LY294002, inhibited in a dose-dependent manner alpha IIb beta 3-mediated platelet spreading on fibrinogen having no effect on platelet spreading on IgG. Both inhibitors also completely abolished alpha IIb beta 3-mediated pp125FAK phosphorylation but not pp72syk phosphorylation. Furthermore, Fc gamma RII- and thrombin-induced pp125FAK phosphorylation were not affected by wortmannin and LY294002. Finally, the PI 3-kinase inhibitors' effect on alpha IIb beta 3-mediated spreading and pp125FAK phosphorylation was reversed by phorbol ester treatment. These results establish that the role of PI 3-kinase relative to pp125FAK phosphorylation in platelets is receptor type-specific yet essential for alpha IIb beta 3-mediated cell spreading and pp125FAK phosphorylation.

Androstadienes↗

Optical imaging reveals reduced seizure spread and propagation velocities in aged rat brain in vitro.

Old age is the most common time for patients to develop epileptic seizures, and due to their frequent unusual clinical presentation the diagnosis of epilepsy is often delayed in the elderly. It is as yet unknown if pronounced alterations in the plastic properties of aging nervous tissue contribute to these phenomena. We employed a non-lesional in vitro epilepsy model to study seizure susceptibility, spread pattern, and propagation velocities in combined hippocampal-entorhinal cortex slices of aged rats and controls using electrophysiological methods and imaging of intrinsic optical signals. In aged animals we saw a less extensive spread of seizure-like events into areas adjacent to the region of onset of activity and a decreased spread velocity in various anatomical regions. In addition, both the activity-dependent shrinkage of the extracellular space (ECS)-volume and the extracellular K(+) concentration were significantly reduced compared to controls. The results of this study are consistent with the clinical observation that epileptic seizures in the elderly have a reduced tendency to spread. In addition, our data suggest that in the absence of structural lesions seizure susceptibility in the aging brain is not increased.

Aging↗

HLA-DP: a class II restriction molecule involved in epitope spreading during the development of multiple sclerosis.

Multiple sclerosis (MS) is an autoimmune demyelinating disease of the central nervous system. It is widely believed that complex polygenic inheritance patterns involving HLA-DR and -DQ class II genes contribute to MS susceptibility, and current evidence indicates that disease risk vs disease outcome may be associated with distinctly different HLA class II alleles. We have recently shown that the early development of MS is accompanied by an extensive plasticity of myelin self-recognition with the acquisition of neo-autoreactivity, or epitope spreading, as a prominent feature. Although we did not observe a common determinant recognized by patients sharing identical HLA-DR or -DQ class II alleles, we did observe epitope spreading to the p50-63 determinant of myelin proteolipid protein (PLP) in two study subjects showing complete disparity at HLA-DR and -DQ but identity at the HLA-DP allele DPB1*0301. In the present study we show that self-recognition during the early stages in the development of MS involves HLA-DP class II restricted responses to the PLP 50-63 spreading determinant. Our results suggest that self-presentation by HLA-DP may play an important role in epitope spreading and in the propagation of self-recognition during the clinical progression of MS.

Adult↗

Charge displacement by adhesion and spreading of a cell.

The potentiostatic control of surface charge density and interfacial tension of an electrode immersed in an aqueous electrolyte solution offers a possibility for direct studies of non-specific interactions in cell adhesion. Unicellular marine alga, Dunaliella tertiolecta (Chlorophyceae) of micrometer size and flexible cell envelope was used as a model cell and 0.1 M NaCl as supporting electrolyte. The dropping mercury electrode acted as in situ adhesion sensor and the electrochemical technique of chronoamperometry allowed measurement of the spread cell-electrode interface area and the distance of the closest approach of a cell. The adhesion and spreading of a single cell at the mercury electrode causes a displacement of counter-ions from the electrical double layer over a broad range of the positive and negative surface charge densities (from +16.0 to -8.2 microC/cm2). The flow of compensating current reflects the dynamics of adhesive contact formation and subsequent spreading of a cell. The adhesion and spreading rates are enhanced by the hydrodynamic regime of electrode's growing fluid interface. The distance of the closest approach of an adherent cell is smaller or equal to the distance of the outer Helmholz plane within the electrical double layer, i.e. 0.3-0.5 nm. There is a clear evidence of cell rupture for the potentials of maximum attraction as the area of the contact interface exceeded up to 100 times the cross-section area of a free cell.

Cell Adhesion↗

Neon colour spreading in three-dimensional illusory objects in humans.

We studied whether neon spreading can be induced within three-dimensional illusory triangles. Kanizsa triangles were induced by black pacman disks consisting of red sectors with curved sides. Viewing our stimuli monocularly produced two-dimensional illusory contours and surfaces as well as neon spreading in each figure. Triangles appeared concave or convex under stereoscopical viewing. Neon colour spreading was induced within illusory figures bending in three-dimensional space, suggesting that neural contour completion and surface filling-in interact across depth. Surprisingly, neon spreading was induced above the intervening surface even when the inducers were below the surface. Neon colour and illusory configuration were preserved behind the intervening surface only when it appeared transparent.

Adolescent↗

Engagement of the NG2 proteoglycan triggers cell spreading via rac and p130cas.

Cells that express the NG2 proteoglycan will spread on surfaces coated with monoclonal antibodies against this membrane-spanning protein. On surfaces coated with the N143 monoclonal antibody, this cell spreading occurs by extension of lamellipodia, suggesting that activation of the small GTPase rac is involved in the observed morphological change. Support for this hypothesis comes from the finding of increased levels of GTP-bound rac in cells spreading on N143-coated surfaces. Furthermore, lamellipodia extension is blocked by transfection of cells with the dominant negative rac construct N17rac, but not by transfection with N17cdc42. Formation of lamellipodia on the N143-coated surfaces is also inhibited by transfection of the dominant negative CasdeltaSD construct. This result implicates p130cas as an additional functional player in NG2-mediated cell spreading.

Antigens↗

Decrease in plasma low-density lipoprotein cholesterol, apolipoprotein B, cholesteryl ester transfer protein, and oxidized low-density lipoprotein by plant stanol ester-containing spread: a randomized, placebo-controlled trial.

OBJECTIVE: The ester of plant stanols significantly reduces plasma levels of total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) in Western people. Effects of plant stanol ester-containing spread on plasma levels of TC, LDL-C, and apolipoprotein B (apoB) were studied in a randomized, placebo-controlled trial in Japanese subjects whose diet is low in fat and cholesterol. The effects of plant stanol ester on plasma levels of arteriosclerosis-promoting factors, namely remnants of triacylglycerol (TG)-rich lipoproteins, cholesteryl ester transfer protein (CETP), and oxidized LDL (Ox-LDL), were also studied. The assessment of safety was also made. METHODS: One hundred and five healthy volunteers were assigned randomly to one of three groups: placebo spread (n = 35), 2 g/d of plant stanol (3.4 g of stanol ester; n = 34), and 3 g/d of plant stanol (5.1 g of stanol ester; n = 36). Plasma levels of lipids were measured at start of the study, at 2 and 4 wk (end of trial), and at 8 wk (+4 wk). Plasma apoproteins, cholesterol in remnant-like particles which are equivalent to remnants of TG-rich lipoproteins (RLP-C), CETP mass, and Ox-LDL were measured at the beginning and the end of the trial. Plasma levels of plant steroids and fat-soluble vitamins were also measured for the assessment of safety. RESULTS: Background and dietary composition did not differ among groups. Plasma levels of TC, LDL-C, apoB, apoE, CETP mass, and Ox-LDL were reduced significantly by 6.5%, 9.6%, 8.3%, 4.5%, 6.1%, and 20%, respectively, in the 2 g/d plant stanol group. Plasma levels of TC, LDL-C, apoB, CETP mass, and Ox-LDL were decreased significantly by 5.5%, 7.3%, 5.6%, 3.3%, and 19%, respectively, in the 3 g/d plant stanol group. Plasma levels of plant stanols, plant sterols, retinol, beta-carotene, and alpha-tocopherol did not change in any group, but levels of campestanol increased and alpha-tocopherol decreased slightly in the sitostanol groups. CONCLUSION: Plasma levels of TC and LDL-C were significantly reduced by the plant stanol ester-containing spread. The smaller reduction than in Western studies and the lack of dose dependency in this study might be due to the different basal diets. We concluded that plant stanol ester-containing spread is efficacious in reducing plasma LDL-C, apoB, CETP, and Ox-LDL and that 2 g/d plant stanol is adequate for Japanese people. No significant side effects were observed in any group.

Apolipoproteins B↗

Modulation of adhesion, spreading and cytoskeleton organization of 3T3 fibroblasts by sulfonic groups present on polymer surfaces.

The early phase of 3T3 fibroblast interaction with sulfonated styrene copolymer surfaces, of two sulfonic group densities and thus of differing wettability, was studied. The sulfonic groups present on copolymer surfaces affected the behaviour of cells, i.e. they stimulated cell adhesion, activated cell spreading and influenced cytoskeleton reorganization. The relative number of adhering cells correlated, while the number of spreading cells inversely correlated, with the surface density of sulfonic groups. Cell shape and the pattern of distribution of F-actin, alpha-actinin and vinculin in the interacting cells also depend on the surface density of sulfonic groups. On surfaces of high sulfonic group density, highly polarized cells were observed with F-actin bundles. On surfaces of low sulfonic group density, the cells spread with a square-like morphology with F-actin organized in stress fibres. In contrast, the cells spread poorly on nonsulfonated surfaces and cell adhesion was unaffected by surface wettability. The contribution of alpha(5)beta(1), alpha(4), and beta(5)integrins to the cell interaction with fibronectin (FN) and vitronectin (VN) adsorbed from serum-containing medium on polymer surfaces was examined. Our results suggest that surface sulfonic groups influence the conformation of FN and VN adsorbed on polymer surfaces and, in turn, determine the integrins that are involved in cell adhesion.

3T3 Cells↗

Recurrent lower gingival squamous cell carcinoma spreading along the pathway of the inferior alveolar nerve.

Computed tomography images showed recurrent lower gingival squamous cell carcinoma spreading from the mandibular foramen to the middle cranial fossa. This extensive tumor spread followed exactly the same anatomic pathway as the inferior alveolar nerve. Histopathologic examination revealed squamous cell carcinoma. Tumor spread was thought to be caused by perineural invasion. This case demonstrates computed tomography features of perineural tumor spread along the inferior alveolar nerve.

Carcinoma, Squamous Cell↗