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Results for “Cell Surface Extensions”

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PubMed2003

Reelin signaling directly affects radial glia morphology and biochemical maturation.

Radial glial cells are characterized, besides their astroglial properties, by long radial processes extending from the ventricular zone to the pial surface, a crucial feature for the radial migration of neurons. The molecular signals that regulate this characteristic morphology, however, are largely unknown. We show an important role of the secreted molecule reelin for the establishment of radial glia processes. We describe a significant reduction in ventricular zone cells with long radial processes in the absence of reelin in the cortex of reeler mutant mice. These defects were correlated to a decrease in the content of brain lipid-binding protein (Blbp) and were detected exclusively in the cerebral cortex, but not in the basal ganglia of reeler mice. Conversely, reelin addition in vitro increased the Blbp content and process extension of radial glia from the cortex, but not the basal ganglia. Isolation of radial glia by fluorescent-activated cell sorting showed that these effects are due to direct signaling of reelin to radial glial cells. We could further demonstrate that this signaling requires Dab1, as the increase in Blbp upon reelin addition failed to occur in Dab1-/- mice. Taken together, these results unravel a novel role of reelin signaling to radial glial cells that is crucial for the regulation of their Blbp content and characteristic morphology in a region-specific manner.

Animals↗

PubMed1983

Ultrastructure of cartilage from young adult sea lamprey, Petromyzon marinus L: a new type of vertebrate cartilage.

Ultrastructural observations of cartilage from adult sea lamprey, Petromyzon marinus, reveal a highly cellular cartilage with an unusual extracellular matrix. The avascular cartilage is surrounded by a vascular perichondrium, which consists of dense connective tissue containing fibroblasts, collagen fibrils, and microfibrils. The cells (chondrocytes) vary in morphology in different parts of the cartilage in a way that may reflect their state of activity. Chondrocytes within the peripheral cartilage contain tubulo-vesicular structures along the cell surface, an extensive lamellar rough endoplasmic reticulum, and a well-developed Golgi complex with associated vesicles and vacuoles. The presence of material within the Golgi elements that resembles components of the extracellular matrix suggests the involvement of the peripheral chondrocytes in the synthesis and secretion of the matrix components. Chondrocytes within the central cartilage are hypertrophied and contain a pale cytoplasm with a reduced number of organelles that are widely spaced throughout the cell. The appearance of the organelles within these cells suggests that they are not as actively involved in the production of the matrix as those of the peripheral cartilage. The extracellular matrix consists of a dense network of randomly arranged, branched, noncollagenous matrix fibrils 15-40 nm in diameter and varying amounts of electron-dense matrix granules. Due to the unique nature of its extracellular matrix, the cartilage of the lamprey cannot be likened to any of the known vertebrate cartilages and, therefore, must be considered a new type of vertebrate cartilage.

Animals↗

PubMed2006

Integrin alpha5beta1 and fibronectin regulate polarized cell protrusions required for Xenopus convergence and extension.

BACKGROUND: Integrin recognition of fibronectin is required for normal gastrulation including the mediolateral cell intercalation behaviors that drive convergent extension and the elongation of the frog dorsal axis; however, the cellular and molecular mechanisms involved are unclear. RESULTS: We report that depletion of fibronectin with antisense morpholinos blocks both convergent extension and mediolateral protrusive behaviors in explant preparations. Both chronic depletion of fibronectin and acute disruptions of integrin alpha5beta1 binding to fibronectin increases the frequency and randomizes the orientation of polarized cellular protrusions, suggesting that integrin-fibronectin interactions normally repress frequent random protrusions in favor of fewer mediolaterally oriented ones. In the absence of integrin alpha5beta1 binding to fibronectin, convergence movements still occur but result in convergent thickening instead of convergent extension. CONCLUSIONS: These findings support a role for integrin signaling in regulating the protrusive activity that drives axial extension. We hypothesize that the planar spatial arrangement of the fibrillar fibronectin matrix, which delineates tissue compartments within the embryo, is critical for promoting productive oriented protrusions in intercalating cells.

Animals↗

PubMed2006

Feeding in a calcareous sponge: particle uptake by pseudopodia.

Sponges are considered to be filter feeders like their nearest protistan relatives, the choanoflagellates. Specialized "sieve" cells (choanocytes) have an apical collar of tightly spaced, rodlike microvilli that surround a long flagellum. The beat of the flagellum is believed to draw water through this collar, but how particles caught on the collar are brought to the cell surface is unknown. We have studied the interactions that occur between choanocytes and introduced particles in the large feeding chambers of a syconoid calcareous sponge. Of all particles, only 0.1-microm latex microspheres adhered to the collar microvilli in large numbers, but these were even more numerous on the choanocyte surface. Few large particles (0.5- and 1.0-microm beads and bacteria) contacted the collar microvilli; most were phagocytosed by lamellipodia at the lateral or apical cell surface, and clumps of particles were engulfed by pseudopodial extensions several micrometers from the cell surface. Although extensions of the choanocyte apical surface up to 16 microm long were found, most were 4 microm long, twice the height of the collar microvilli. These observations offer a different view of particle uptake in sponges, and suggest that, at least in syconoid sponges, uptake of particles is less dependent on the strictly sieving function of the collar microvilli.

Animals↗

PubMed2002

Hippocampal astrocytes in situ exhibit calcium oscillations that occur independent of neuronal activity.

Results presented in this study indicate that a large subpopulation (approximately 65%) of hippocampal astrocytes in situ exhibit calcium oscillations in the absence of neuronal activity. Further, the spontaneous oscillations observed within individual hippocampal astrocytes generally developed asynchronously throughout the astrocyte's fine processes and occasionally spread through a portion of that astrocyte as a calcium wave but do not appear to spread among astrocytes as an intercellular calcium wave. Bath application of cyclopiazonic acid and injection of individual astrocytes with heparin blocked astrocytic calcium oscillations. Application of tetrodotoxin or incubation of slices with bafilomycin A1 had no effect on astrocytic calcium oscillations but did block evoked and spontaneous postsynaptic currents measured in CA1 pyramidal neurons. Application of a cocktail of antagonists for metabotropic glutamate receptors and purinergic receptors had no effect on the astrocytic calcium oscillations but blocked the ability of purinergic and metabotropic glutamatergic agonists to increase astrocytic calcium levels. These results indicate that the spontaneous calcium oscillations observed in hippocampal astrocytes in situ are mediated by IP3 receptor activation, are not dependent on neuronal activity, and do not depend on activation of metabotropic glutamate receptors or purinergic receptors. To our knowledge, this is the first demonstration that astrocytes in situ exhibit intrinsic signaling. This finding supports the hypothesis that astrocytes, independent of neuronal input, may act as pacemakers to modulate neuronal activity in situ.

Animals↗

PubMed1994

Ultracytochemical study of multiple dose effect of monastral blue uptake by equine pulmonary intravascular macrophages (PIMs).

The pulmonary intravascular macrophages (PIMs) of the ponies possess all the characteristics of a fully differentiated resident macrophage, which forms adhesive plaques with the capillary-endothelial cells. In addition, it has a unique surface coat which does not conform to the concept of a carbohydrate rich membrane bound glycocalyx generally associated with macrophages. We studied the responses of these cells especially in the context of its globular surface coat to multiple doses of MB intravenously: 0.2 ml/kg body copper tracer substance. The ponies were treated with three doses of MB intravenously: 0.2 ml/kg body weight on day 0, 0.1 ml/kg at 48 h and a 0.05 ml/kg at 96 h after the first dose. The examination of lung tissue at the ultrastructural level revealed internalization of MB following complexing with the surface coat globules along the entire cell surface. The cell surface was extensively ruffled in the form of several lamellipods. The MB-globule complex was internalized mostly at the coated pits whereas concurrent internalization of nascent globules of the coat took place at the ruffled surface. In the former, the internalized material was seen in small, medium and large phagolysosomes which stained intensely for acid phosphatase, whereas in latter case enzyme activity was entirely absent in all the nascent globules carrying endosomes. Simultaneously there was hypertrophy of the Golgi complex, which showed intense staining for acid phosphatase. Relative to the Golgi response, acid phosphatase-positive tubular lysosomes in close association with extensive bundles of microtubules were conspicuous, thus adding to the battery of cell organelles of secretory pathway.(ABSTRACT TRUNCATED AT 250 WORDS)

Acid Phosphatase↗

PubMed2005

Like cops on the beat: the active role of resting microglia.

Microglia form the first line of defence for the neural parenchyma. But do these cells pursue an active role in the normal brain, or do they become activated only after injury? Two papers published recently by Nimmerjahn et al. and Davalos et al. used in vivo two-photon laser-scanning microscopy reveal that the fine branches of 'resting' microglia are highly mobile, and provide extensive and continuous surveillance of their cellular environment. These moving branches show a rapid chemotactic response to tissue injury that depends on purine receptors and connexin hemichannels, and they appear to take cues from surrounding astrocytes both in the normal and the injured brain.

Animals↗

PubMed1980

Quantitative analysis of exocytosis and endocytosis in the hydroosmotic response of toad bladder.

This study concerns the timing and magnitude of exocytosis and endocytosis in the granular cells of toad bladder during the hydroosmotic response to antidiuretic hormone. Granule exocytosis at the luminal cell surface is extensive within 5 min of the administration of a physiological dose of hormone. Hydroosmosis becomes detectable during this time period. The amount of membrane added to the luminal surface by exocytosis during 60 min of exposure to hormone can be of the same order of magnitude as the extent of the luminal plasma membrane. Endocytosis, demonstrated by peroxidase uptake from the luminal surface, becomes extensive during the period 15-45 min after hormone administration. Thus, maximal endocytic activity occurs later than the period of most extensive exocytosis and seems to correlate with the onset of the decline in water movement. The amount of membrane retrieved from the luminal surface by endocytosis during 60 min of stimulation is at least three quarters of that added by exocytosis. The bulk membrane movement in ADH stimulated preparations does not require the presence of an osmotic gradient. Colchicine inhibits the hydroosmotic response, the exocytosis of granules, and endocytosis at the luminal surface. These results strengthen our view that the bulk circulation of membrane at the cell surface, via exocytosis and endocytosis, is closely related to the permeability changes occurring at the surface.

Animals↗

PubMed2002

Transforming growth factor-beta-induced mobilization of actin cytoskeleton requires signaling by small GTPases Cdc42 and RhoA.

Transforming growth factor-beta (TGF-beta) is a potent regulator of cell growth and differentiation in many cell types. The Smad signaling pathway constitutes a main signal transduction route downstream of TGF-beta receptors. We studied TGF-beta-induced rearrangements of the actin filament system and found that TGF-beta 1 treatment of PC-3U human prostate carcinoma cells resulted in a rapid formation of lamellipodia. Interestingly, this response was shown to be independent of the Smad signaling pathway; instead, it required the activity of the Rho GTPases Cdc42 and RhoA, because ectopic expression of dominant negative mutant Cdc42 and RhoA abrogated the response. Long-term stimulation with TGF-beta 1 resulted in an assembly of stress fibers; this response required both signaling via Cdc42 and RhoA, and Smad proteins. A known downstream effector of Cdc42 is p38(MAPK); treatment of the cells with the p38(MAPK) inhibitor 4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(pyridyl)1H-imidazole (SB203580), as well as ectopic expression of a kinase-inactive p38(MAPK), abrogated the TGF-beta-induced actin reorganization. Moreover, treatment of cells with the inhibitors of the RhoA target-protein Rho-associated coiled-coil kinase (+)-R-trans-4-(aminoethyl)-N-(4-pyridyl) cyclohexanecarboxamide (Y-27632) and 1-5(-isoquinolinesulfonyl)homopiperazine (HA-1077), as well as ectopic expression of kinase-inactive Rho coiled-coil kinase-1, abrogated the TGF-beta 1-induced formation of stress fibers. Collectively, these data indicate that TGF-beta-induced membrane ruffles occur via Rho GTPase-dependent pathways, whereas long-term effects require cooperation between Smad and Rho GTPase signaling pathways.

Actins↗

PubMed2001

Rapid formation and remodeling of postsynaptic densities in developing dendrites.

The dynamics of postsynaptic density (PSD) formation and remodeling were investigated in live developing hippocampal tissue slices. Time lapse imaging of transfected neurons expressing GFP-tagged PSD95, a prominent PSD protein, revealed that up to 40% of PSDs in developing dendrites are structurally dynamic; they rapidly (<15 min) appear or disappear, but also grow, shrink and move within shafts and spines. New spines containing PSDs were formed by conversion of dynamic filopodia-like spine precursors in which PSDs appeared de novo, or by direct extension of spines or spine precursors carrying preformed PSDs from the shaft. PSDs are therefore highly dynamic structures that can undergo rapid structural alteration within dendrite shafts, spines and spine precursors, permitting rapid formation and remodeling of synaptic connections in developing CNS tissues.

Animals↗

PubMed2003

Epithelioid ependymoma: a new variant of ependymoma: report of three cases.

OBJECTIVE: To describe the pathological features of three very similar and unusual primary central nervous system tumors that are not readily recognized as conventional ependymomas but which, by ultrastructural examination, have an ependymomatous character. METHODS: Three distinctive tumors were found in a review of our files for cases of ependymoma. In each case, hematoxylin and eosin-stained sections were reviewed, and immunostains for epithelial membrane antigen, cytokeratin, vimentin, and glial fibrillary acidic protein were performed on formalin-fixed, paraffin-embedded sections. Electron microscopy was performed in each case. RESULTS: The tumors had a diffuse myxoid background, often containing tightly clustered cells that mimicked multinucleated giant cells, but lacking perivascular pseudorosettes or central lumen rosettes. Glial fibrillary acidic protein and vimentin immunostains did not reveal perivascular processes. Epithelial membrane antigen immunostains showed a dot-like cytoplasmic immunoreactivity in some cell clusters in two of the three cases. Cytokeratin was negative in all three cases. However, ultrastructurally, the cells of each tumor had extensive surface microvilli; the giant cell-like clusters had cells with extensive close appositions, some junctions, and, in two cases, lumina with microvilli. Two of the patients were adults (both with temporal lobe tumors), and one patient was 13 years old and had a cervical spinal cord intramedullary tumor. Each tumor was sharply circumscribed from adjacent central nervous system tissue but was not encapsulated. One of the cases in an adult was mitotically highly active; this tumor recurred locally 4 years after initial gross total excision. CONCLUSION: These tumors are unusual variants of ependymoma. This pattern of ependymoma is sufficiently distinctive to be recognized in hematoxylin and eosin stains once the architecture of the epithelioid clusters is appreciated.

Adolescent↗

PubMed2004

DISTORTED2 encodes an ARPC2 subunit of the putative Arabidopsis ARP2/3 complex.

Arabidopsis trichomes are unicellular, branched structures that have highly constrained requirements for the cytoskeleton. The 'distorted group' genes function downstream from microtubule-based branch initiation, and are required during the actin-dependent phase of polarized stalk and branch expansion. Of the eight known 'distorted group' genes, a subset encode homologs of ARP2/3 complex subunits. In eukaryotic cells, the seven-protein ARP2/3 complex nucleates actin filament networks that push on the plasma membrane and organelles. In plants cells, the existence and function of an ARP2/3 complex is unclear. In this paper, we report that DISTORTED2 (DIS2) encodes a paralogue of the ARP2/3 complex subunit ARPC2. DIS2 has ARPC2 activity, based on its ability to rescue the growth defects of arpc2 (arc35Delta) null yeast cells. Like known ARPC2s, DIS2 physically interacts with ARPC4. Mutations in DIS2 cause a distorted trichome phenotype, defects in cell-cell adhesion, and a modest reduction in shoot FW. The actin cytoskeleton in dis2 trichomes is extensive, but developing branches fail to generate and maintain highly organized cytoplasmic actin bundles.

Actin-Related Protein 2↗

PubMed2004

Tri-dimensional morphometric analysis of astrocytic processes with high voltage electron microscopy of thick Golgi preparations.

A characteristic feature of the astrocytic processes is to assume the form of shin sheets or lamellate coverings of other brain constituents. We analyzed the extensive and finely divided processes of the protoplasmic astrocyte in the molecular layer of the rat dentate gyrus by means of computer electron tomography and stereo-photogrammetry using tilted high voltage electron microscope images of thick Golgi preparations. The surface area and volume of the astrocytic processes were measured and the surface/volume ratios were estimated. The surface/volume ratios of astrocytic processes in the neuropile ranged from 18.9 to 33.0 per microm, and the mean value was 26.2+/-5.0 per microm. The values were roughly comparable to those previously reported for the microdomain of Bergmann glia cell terminal processes in the rat cerebellum, which were estimated from reconstructions using thin serial section electron microscope images. The large surface to volume ratio of the astrocytic processes in the neuropile resulted from the lamellar nature of the processes interposed between other cellular elements, and may reflect the functional activities of the astrocyte. The results suggest the usefulness of the electron tomography and stereo-photogrammetry for three-dimensional morphometrical analysis of the astrocytic processes, although both techniques can be expected to be refined further in order to provide more precise measurements of these complicated processes.

Animals↗

PubMed1988

The rate and topography of cell wall synthesis during the division cycle of Escherichia coli using N-acetylglucosamine as a peptidoglycan label.

The rates of synthesis of peptidoglycan and protein during the division cycle of Escherichia coli were measured by the membrane elution technique using cells differentially labelled with N-acetylglucosamine and leucine. During the first part of the division cycle the ratio of the rates of protein and peptidoglycan synthesis was constant. The rate of peptidoglycan synthesis, relative to the rate of protein synthesis, increased during the latter part of the division cycle. These results support a simple, bipartite model of cell surface increase in rod-shaped cells. Prior to the start of constriction the cell surface increases only by lateral wall extension. After cell constriction starts, the cell surface increases by both lateral wall and pole growth. The increase in surface area is partitioned between the lateral wall and the pole so that the volume of the cell increases exponentially. No variation in cell density occurs, because the increase in surface allows a continuous exponential increase in cell volume that accommodates the exponential increase in cell mass. The results are consistent with the constant density of the growing cell and the surface stress model for the regulation of cell surface synthesis. In addition, the elution pattern suggests that the membrane elution method does work by having the cells effectively bound to the membrane by their poles.

Acetylglucosamine↗

PubMed2004

Over-expression of the p110beta but not p110alpha isoform of PI 3-kinase inhibits motility in breast cancer cells.

Phosphoinositide 3-kinase (PI 3-kinase) activity is required for growth factor-induced cytoskeletal regulation and cell migration. We previously found that in MTLn3 rat adenocarcinoma cells, EGF-stimulated induction of actin barbed ends and lamellipod extension specifically requires the p85/p110alpha isoform of PI 3-kinase. To further characterize signaling by distinct PI 3-kinase isoforms, we have developed MTLn3 cells that transiently or stably overexpress either p110alpha or p110beta. Transient overexpression of p110beta inhibited EGF-stimulated lamellipod extension, whereas p110alpha-transfected cells showed normal EGF-stimulated lamellipod extension. Similar results were obtained by overexpression of kinase-dead p110beta, suggesting that effects on cytoskeletal signaling were due to competition with p85/p110alpha complexes. Stable overexpression of p110alpha appeared to be toxic, based on the difficulty in obtaining stable overexpressing clones. In contrast, cells expressing a 2-fold increase in p110beta were readily obtainable. Interestingly, cells stably expressing p110beta showed a marked inhibition of EGF-stimulated lamellipod extension. Using computer-assisted analysis of time-lapse images, we found that overexpression of p110beta caused a nearly complete inhibition of motility. Cells overexpressing p110beta showed normal activation of Akt and Erk, suggesting that overall PI 3-kinase signaling was intact. A chimeric p110 molecule containing the p85-binding and Ras-binding domains of p110alpha and the C2, helical, and kinase domains of p110beta, was catalytically active yet also inhibited EGF-stimulated lamellipod extension. These data highlight the differential signaling by distinct p110 isoforms. Identification of effectors that are differently regulated by p110alpha versus p110beta will be important for understanding cell migration and its role in metastasis.

Adenocarcinoma↗

PubMed2006

P21-activated kinase 1: convergence point in PDGF- and LPA-stimulated collagen matrix contraction by human fibroblasts.

Fibroblast three-dimensional collagen matrix culture provides a tissue-like model that can be used to analyze cell form and function. The physiological agonists platelet-derived growth factor (PDGF) and lysophosphatidic acid (LPA) both stimulate human fibroblasts to contract floating collagen matrices. In this study, we show that the PDGF and LPA signaling pathways required for matrix contraction converge on p21-activated kinase 1 (PAK1) and its downstream effector cofilin1 and that contraction depends on cellular ruffling activity, rather than on the protrusion and retraction of cellular dendritic extensions. We also show that, depending on the agonist, different Rho effectors cooperate with PAK1 to regulate matrix contraction, Rho kinase in the case of PDGF and mDia1 in the case of LPA. These findings establish a unified framework for understanding the cell signaling pathways involved in fibroblast contraction of floating collagen matrices.

Adaptor Proteins, Signal Transducing↗

PubMed2001

The role of matrix metalloproteinase genes in glioma invasion: co-dependent and interactive proteolysis.

Matrix metalloproteinases (MMPs) are cation-dependent endopeptidases which have been implicated in the malignancy of gliomas. It is thought that the MMPs play a critical role in both metastasis and angiogenesis, and that interference with proteases might therefore deter local tumor dissemination and neovascularization. However, the attempt to control tumor-associated proteolysis will rely on better definition of the normal tissue function of MMPs, an area of study still in its infancy in the central nervous system (CNS). Understanding the role of MMP-mediated proteolysis in the brain relies heavily on advances in other areas of molecular neuroscience, most notably an understanding of extracellular matrix (ECM) composition and the function of cell adhesion molecules such as integrins, which communicate knowledge of ECM composition intracellularly. Recently, protease expression and function has been shown to be strongly influenced by the functional state and signaling properties of integrins. Here we review MMP function and expression in gliomas and present examples of MMP profiling studies in glioma tissues and cell lines by RT-PCR and Western blotting. Co-expression of MMPs and certain integrins substantiates the gathering evidence of a functional intersection between the two, and inhibition studies using recombinant TIMP-1 and integrin antisera demonstrate significant inhibition of glioma invasion in vitro. Use of promising new therapeutic compounds with anti-MMP and anti-invasion effects are discussed. These data underline the importance of functional interaction of MMPs with accessory proteins such as integrins during invasion, and the need for further studies to elucidate the molecular underpinnings of this process.

Animals↗

PubMed2004

Antibacterial titanium plate anodized by being discharged in NaCl solution exhibits cell compatibility.

Implant surfaces should be modified to achieve excellent cell compatibility as well as antibacterial activity. Our previous study demonstrated that titanium plates anodized by being discharged in NaCl (Ti-Cl) exhibited high antibacterial activity. Since Ti-Cl was prepared with a NaCl solution, we hypothesized that Ti-Cl would exhibit low toxicity toward cells. The aims of this study were to characterize the surface of Ti-Cl and investigate the cell compatibility (MC3T3-E1 and L929 cells) of Ti-Cl. The results demonstrated that, since the TiCl(3) formed on the Ti-Cl surface was hydrolyzed into HCl, HClO, and TiOH after immersion in pure distilled water, TiCl(3) contributed to the antibacterial activity of Ti-Cl. On the other hand, TiO formed on the Ti-Cl surface enhanced cell extension and cell growth through a larger adsorption of fibronectin compared with the pure titanium control. These findings suggest that antibacterial titanium is a promising material for use in dental implant systems.

3T3 Cells↗