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Neighboring Cells (neighboring + cell)
Selected AbstractsAn in vivo comparison of photoactivatable fluorescent proteins in an avian embryo modelDEVELOPMENTAL DYNAMICS, Issue 6 2007Danny A. Stark Abstract Tracing the lineage or neighbor relationships of cells in a migratory population or deep within an embryo is difficult with current methods. The recent explosion of photoactivatable fluorescent proteins (PAFPs) offers a unique cell labeling tool kit, yet their in vivo performance in intact embryos and applicability have not been thoroughly explored. We report a comparison study of PAGFP, PSCFP2, KikGR, and Kaede analyzed in the avian embryo using confocal and 2-photon microscopy. PAFPs were introduced into the chick neural tube by electroporation and each photoconverted in the neural crest or cells in the neural tube with exposure to 405 nm light, but showed dramatic differences in photoefficiency and photostability when compared at the same 2% laser power. KikGR and Kaede photoconverted with ratios only slightly lower than in vitro results, but cells rapidly photobleached after reaching maximal photoefficiency. PSCFP2 had the lowest photoefficiency and photoconverted nearly 70 times slower than the other dual-color PAFPs tested, but was effective at single-cell marking, especially with 2-photon excitation at 760 nm. The dual-color PAFPs were more effective to monitor cell migratory behaviors, since non-photoconverted neighboring cells were fluorescently marked with a separate color. However, photoconverted cells were limited in all cases to be visually distinguishable for long periods, with PSCFP2 visible from background the longest (48 hr). Thus, photoactivation in embryos has the potential to selectively mark less accessible cells with laser accuracy and may provide an effective means to study cell,cell interactions and short-term cell lineage in developmental and stem cell biology. Developmental Dynamics 236:1583,1594, 2007. © 2007 Wiley-Liss, Inc. [source] An adaptive resource reservation for vehicular mobile networksINTERNATIONAL JOURNAL OF NETWORK MANAGEMENT, Issue 5 2009I. Ben Hamida This paper presents the time-based bandwidth reservation (TBR) algorithm, suitable for handoff management in cellular systems. TBR is based on real-time measurements of mobile stations (position, velocity and acceleration). The scheme consists in sending reservation requests to the neighboring cells based on an extrapolation of the user's motion. The originality of our approach lies in dynamically adjusting the amount of time for which bandwidth has to be allocated and reserved in a cell. In addition, we propose an optimal channel requests arrangement (CRA) algorithm in order to improve the performance of TBR in terms of resource utilization. Finally, we propose VTBR, an adapted and extended version of TBR for better support of vehicular network specificities where service degradation or forced call termination may occur owing to frequent handoffs. Detailed simulation results for TBR and VTBR schemes and a comparison with the guard channel scheme are presented. The results show that TBR and VTBR can efficiently improve the flow dropping probability. Copyright © 2008 John Wiley & Sons, Ltd. [source] Vesicle traffic through intercellular bridges in DU 145 human prostate cancer cellsJOURNAL OF CELLULAR AND MOLECULAR MEDICINE, Issue 3 2004Cristina Vidulescu Abstract We detected cell-to-cell communication via intercellular bridges in DU 145 human prostate cancer cells by fluorescence microscopy. Since DU 145 cells have deficient gap junctions, intercellular bridges may have a prominent role in the transfer of chemical signals between these cells. In culture, DU 145 cells are contiguous over several cell diameters through filopodial extensions, and directly communicate with adjacent cells across intercellular bridges. These structures range from 100 nm to 5 ,m in diameter, and from a few microns to at least 50,100 ,m in length. Time-lapse imagery revealed that (1) filopodia rapidly move at a rate of microns per minute to contact neighboring cells and (2) intercellular bridges are conduits for transport of membrane vesicles (1,3 ,m in diameter) between adjacent cells. Immunofluorescence detected alpha-tubulin in intercellular bridges and filopodia, indicative of microtubule bundles, greater than a micron in diameter. The functional meaning, interrelationship of these membrane extensions are discussed, along with the significance of these findings for other culture systems such as stem cells. Potential applications of this work include the development of anticancer therapies that target intercellular communication and controlling formation of cancer spheroids for drug testing. [source] Glial connexins and gap junctions in CNS inflammation and diseaseJOURNAL OF NEUROCHEMISTRY, Issue 3 2008Tammy Kielian Abstract Gap junctions facilitate direct cytoplasmic communication between neighboring cells, facilitating the transfer of small molecular weight molecules involved in cell signaling and metabolism. Gap junction channels are formed by the joining of two hemichannels from adjacent cells, each composed of six oligomeric protein subunits called connexins. Of paramount importance to CNS homeostasis are astrocyte networks formed by gap junctions, which play a critical role in maintaining the homeostatic regulation of extracellular pH, K+, and glutamate levels. Inflammation is a hallmark of several diseases afflicting the CNS. Within the past several years, the number of publications reporting effects of cytokines and pathogenic stimuli on glial gap junction communication has increased dramatically. The purpose of this review is to discuss recent observations characterizing the consequences of inflammatory stimuli on homocellular gap junction coupling in astrocytes and microglia as well as changes in connexin expression during various CNS inflammatory conditions. [source] Growth and Differentiation of Osteoblast-Like Cells from Calvaria of Connexin43 Deficient MiceMATERIALWISSENSCHAFT UND WERKSTOFFTECHNIK, Issue 12 2004M. Wiemann Osteoblasten-artige Zellen; Connexin43-defiziente Mäuse; gap junctions; Differenzierung Abstract Extensive cell-cell-coupling via gap junctions has been suspected to play an essential role for osteoblast development. Here, osteoblast-like cells (OBL) from connexin(Cx)43 knock out mice were used to explore the role of Cx43 for osteoblast differentiation. Primary cultures of OBL were derived from calvaria of homozygous (Cx43-/-) and heterozygous (Cx43+/,) knock out mice and also from wild type controls (Cx43+/+). In Cx43-/- OBL Lucifer Yellow dye coupling was largely abolished demonstrating that small molecules could no longer be transferred among neighboring cells. Cx43-/- OBL grew out very slowly from calvarial fragments. Nevertheless their cell density around explants was increased 3-fold vs. controls after 3 weeks. Histochemistry showed that in many Cx43-/- OBL there was an increased alkaline phosphatase activity within the cytoplasm and close to the cell membrane. Mineralization was diminished in Cx43-/- cultures. In heterozygous Cx43+/, OBL all aforementioned effects were less pronounced, pointing to a gene-dosage effect. Data suggest that the loss of Cx43 indirectly impairs the osteoblastic phenotype, e.g. by disturbing cellular functions such as motility and/or secretion. If this holds true, all parameters in the interphase of enosseous implants which lower gap junction expression will also affect bone regeneration. Wachstum und Differenzierung von Osteoblasten-artigen Zellen aus Kalvarien Connexin43-defizienter Mäuse Es wurde oft vermutet, dass die ausgeprägte Zell-Zell-Kopplung von Osteoblasten durch gap junctions eine besondere Rolle für die Differenzierung der gekoppelten Zellen spielt. In dieser Arbeit wurden daher Osteoblasten-artige Zellen (OBL) aus Connexin43 (Cx43) knock out Mäusen benutzt, um die Bedeutung von Cx43-gap-junction-Kanälen für die Differenzierung von Osteoblasten zu untersuchen. Die dafür notwendigen OBL-Primärkulturen wurden aus Calvarienfragmenten von homozygoten (Cx43-/-) und heterozygoten (Cx43+/,) knock out Mäusen sowie aus Wildtyp-Mäusen gewonnen. In Cx43-/- OBL war die Lucifer Yellow-Farbstoffkopplung weitgehend aufgehoben. Dieser Befund zeigt, dass Moleküle ,600 D zwischen Cx43-/- Zellen kaum noch ausgetauscht werden können. Cx43-/- Zellen wuchsen vergleichsweise langsam aus ihren Calvarienfragmenten aus. Dennoch erreichten diese Kulturen nach 3 Wochen eine im Vergleich zur Kontrolle 3fach höhere Zelldichte. Histochemisch zeigte sich, dass in Cx43-/- Zellen die alkalische Phosphatase-Aktivität im Zytoplasma und besonders im Bereich der Zellmembran erhöht war. Die Mineralisierung war hingegen herabgesetzt. In heterozygoten Cx43+/, OBL waren alle genannten Effekte intermediär ausgeprägt, was auf einen Gen-Dosis-Effekt deutet. Insgesamt legen die Befunde nahe, dass der Verlust von Cx43 die Ausprägung des osteoblastären Phänotyps, z.,B. durch eine Behinderung der Zellbeweglichkeit und/oder der Sekretion beeinträchtigt. Daher dürften alle Parameter, die die Expression von Cx43 im Interphase eines enossalen Implantats stören, die Knochenregeneration behindern. [source] Proapoptotic Nitric Oxide Production in Amyloid , Protein-Treated Cerebral Microvascular Endothelial CellsMICROCIRCULATION, Issue 2 2007CHIWAKA KIMURA ABSTRACT Objective: The objective of this study was to investigate the effects of amyloid , protein (A,) on cerebral microvascular endothelium, and their possible involvement in A,-induced apoptosis in the neighboring cells. Methods: Cultured bovine brain microvascular endothelial cells (BBECs) were incubated with A, for 24 h. Production of nitric oxide (NO) was assessed by nitric oxide-sensitive fluorescent dye, DAF-2, and the expression of NO synthase (NOS) proteins was examined by Western blotting. Effects of A,-treated microvascular endothelium on the DNA damage of the neighboring cells were assessed by single-cell gel electrophoresis. Results: A, increased the expression of iNOS protein, but did not affect eNOS and nNOS expressions in BBECs. A,-treated BBECs showed spontaneous NO production in the presence of L-arginine. The neural cell line PC12 showed marked apoptosis after being co-cultured with A,-treated BBECs for 48 h, and the apoptosis was as potent as that induced by the inflammatory stimuli lipopolysaccharide and interferon-,. The DNA damage of PC12 cells evoked by co-culture with A,-treated BBECs was prevented by L- NG -nitroarginine methyl ester, an inhibitor of NOS. Conclusions: These results indicate that A, induces the expression of iNOS in BBECs, and that microvascular endothelium-derived NO may induce apoptosis in neighboring neural cells. [source] The morphogenesis of lobed plant cells in the mesophyll and epidermis: organization and distinct roles of cortical microtubules and actin filamentsNEW PHYTOLOGIST, Issue 3 2005Emmanuel Panteris Summary The morphogenesis of lobed plant cells has been considered to be controlled by microtubule (MT) and/or actin filament (AF) organization. In this article, a comprehensive mechanism is proposed, in which distinct roles are played by these cytoskeletal components. First, cortical MT bundles and, in the case of pavement cells, radial MT arrays combined with MT bundles determine the deposition of local cell wall thickenings, the cellulose microfibrils of which copy the orientation of underlying MTs. Cell growth is thus locally prevented and, consequently, lobes and constrictions are formed. Arch-like tangential expansion is locally imposed at the external periclinal wall of pavement cells by the radial arrangement of cellulose microfibrils at every wall thickening. Whenever further elongation of the original cell lobes occurs, AF patches assemble at the tips of growing lobes. Intercellular space formation is promoted or prevented by the opposite or alternate, respectively, arrangement of cortical MT arrays between neighboring cells. The genes that are possibly involved in the molecular regulation of the above morphogenetic procedure by MT and AF array organization are reviewed. [source] Proteomic profiling of exosomes: Current perspectivesPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 19 2008Richard J. Simpson Professor Abstract Exosomes are 40,100,nm membrane vesicles of endocytic origin secreted by most cell types in vitro. Recent studies have shown that exosomes are also found in vivo in body fluids such as blood, urine, amniotic fluid, malignant ascites, bronchoalveolar lavage fluid, synovial fluid, and breast milk. While the biological function of exosomes is still unclear, they can mediate communication between cells, facilitating processes such as antigen presentation and in trans signaling to neighboring cells. Exosome-like vesicles identified in Drosophila (referred to as argosomes) may be potential vehicles for the spread of morphogens in epithelia. The advent of current MS-based proteomic technologies has contributed significantly to our understanding of the molecular composition of exosomes. In addition to a common set of membrane and cytosolic proteins, it is becoming increasingly apparent that exosomes harbor distinct subsets of proteins that may be linked to cell-type associated functions. The secretion of exosomes by tumor cells and their implication in the transport and propagation of infectious cargo such as prions and retroviruses such as HIV suggest their participation in pathological situations. Interestingly, the recent observation that exosomes contain both mRNA and microRNA, which can be transferred to another cell, and be functional in that new environment, is an exciting new development in the unraveling exosome saga. The present review aims to summarize the physical properties that define exosomes as specific cell-type secreted membrane vesicles. [source] Regulation of restitution after superficial injury in isolated guinea pig gastric mucosaAPMIS, Issue 4-5 2004ARUN BHOWMIK The immediate response of the gastrointestinal epithelium to superficial (i.e. microscopic) injury is primarily directed towards restoring the disturbed epithelial continuity. Both structural (i.e. cytoskeleton) and humoral (i.e. growth factors and cytokines) involvement in the process has recently been documented. Yet it is unclear whether humoral signaling regulating mucosal recovery after superficial injury is associated with tyrosine phosphorylation, and whether there are other signs of downstream activation of the signaling pathway. To evaluate the effects of exogenous genistein and phorbol-myristate acetate in the assessment of the role of tyrosine receptor-mediated signaling in the immediate repair of gastric mucosa after superficial injury. Guinea pig gastric mucosa was mounted in a Ussing chamber, injured with 1.25 M NaCl, and perfused for 4 h. Simultaneously, potential difference and tissue resistance were recorded. In some sets of experiments the tissue was exposed bilaterally either to genistein in order to inhibit tyrosine receptor-mediated signaling or to 4-phorbol-myristate 13-acetate (PMA) in order to enhance PKC signaling during the 4 h recovery. Phosphotyrosine (PTYR) and protein kinase C (PKC) immunoreactivity were assessed by immunoblotting and by immunohistochemistry. Proliferative activity was determined morphometrically after staining of the tissue for Ki-67 nuclear antigen and expressed as proliferative index (PI). The inhibition of tyrosine kinases with exogenous genistein resulted in a significant decrease of the PTYR and the stimulation of PKC with PMA increased the PTYR. Nevertheless, no change in the PTYR was observed by immunoblotting after superficial injury alone. Several PKC isoenzymes were found in the guinea pig gastric mucosa, including PKC-,, -,, -, and -,. They were unaffected either by the injury or the PMA treatment. The mean PI of tissues subjected to NaCl-injury was higher than that of uninjured control tissues (p<0.05) (n=7). Exposure of tissue to genistein during recovery decreased the PI, while stimulation with PMA increased it (p<0.05 for both) (n=6). Both electrophysiologic and morphologic restitution were sensitive to genistein, but not to PMA. Superficial injury alone does not influence tyrosine phosphorylation to a degree which could be assessed by immunoblotting. Nevertheless, exogenous modulation of tyrosine receptor-mediated signaling results in downstream signaling effects. The injury-associated induction of proliferation is sensitive to modulation of tyrosine phosphorylation and PKC, suggesting that superficial epithelial injury results in endogenous activation of the epithelium, presumably after paracrine stimulation of the neighboring cells. [source] Key Factors in Alzheimer's Disease: ,-amyloid Precursor Protein Processing, Metabolism and Intraneuronal TransportBRAIN PATHOLOGY, Issue 1 2001Thomas A. Bayer During the last years it has become evident that the ,-amyloid (A,) component of senile plaques may be the key molecule in the pathology of Alzheimer's disease (AD). The source and place of the neurotoxic action of A,, however, is still a matter of controversy. The precursor of the ,-amyloid peptide is the predominantly neuronal ,-amyloid precursor protein. We, and others, hypothesize that intraneuronal misregulation of APP leads to an accumulation of A, peptides in intracellular compartments. This accumulation impairs APP trafficking, which starts a cascade of pathological changes and causes the pyramidal neurons to degenerate. Enhanced A, secretion as a function of stressed neurons and remnants of degenerated neurons provide seeds for extracellular A, aggregates, which induce secondary degenerative events involving neighboring cells such as neurons, astroglia and macrophages/microglia. [source] Irradiated fibroblast-induced bystander effects on invasive growth of squamous cell carcinoma under cancer,stromal cell interactionCANCER SCIENCE, Issue 12 2008Noriyuki Kamochi The irradiated fibroblast-induced response of non-irradiated neighboring cells is called ,radiation-induced bystander effect', but it is unclear in non-irradiated human squamous cell carcinoma (SCC) cells. The present study shows that irradiated fibroblasts promoted the invasive growth of T3M-1 SCC cells, but not their apoptosis, more greatly than non-irradiated fibroblasts, using collagen gel invasion assay, immunohistochemistry and Western blot. The number of irradiated fibroblasts decreased to about 30% of that of non-irradiated fibroblasts, but irradiated fibroblasts increased the growth marker ki-67 display of SCC cells more greatly than non-irradiated fibroblasts. Irradiated fibroblasts did not affect the apoptosis marker ss-DNA expression of SCC cells. Irradiated fibroblasts enhanced the display of the following growth-, invasion- and motility-related molecules in SCC cells more greatly than non-irradiated fibroblasts: c-Met, Ras, mitogen-activated protein kinase (MAPK) cascade (Raf-1, MEK-1 and ERK-1/2), matrix metalloproteinase-1 and -9, laminin 5 and filamin A. Irradiated fibroblasts, but not non-irradiated ones, formed irradiation-induced foci (IRIF) of the genomic instability marker p53-binding protein 1 (53BP1) and expressed transforming growth factor-,1 (TGF- ,1). Irradiated fibroblasts in turn enabled SCC cells to enhance 53BP1 IRIF formation more extensively than non-irradiated fibroblasts. Finally, effects of irradiated fibroblasts on growth and apoptosis of another HEp-2 SCC cell type were similar to those of T3M-1. These results suggest that irradiated fibroblasts promotes invasion and growth of SCC cells by enhancement of invasive growth-related molecules above through TGF- ,1-mediated bystander mechanism, in which irradiated fibroblast-induced genomic instability of SCC cells may be involved. (Cancer Sci 2008; 99: 2417,2427) [source] Spontaneous oscillation and mechanically induced calcium waves in chondrocytesCELL BIOCHEMISTRY AND FUNCTION, Issue 2 2006Taisuke Kono Abstract The characteristics of spontaneous calcium (Ca2+) oscillation and mechanically induced Ca2+ waves in articular chondrocytes were studied. In some, but not all, chondrocytes in sliced cartilage and primary cultures, we observed spontaneous oscillation of intracellular Ca2+ that never spread to adjacent cells. In contrast, a mechanical stimulus to a single cell by touching with a glass rod induced an increase of intracellular Ca2+ that spread to neighboring cells in a wave-like manner, even though there was no physical contact between the cells. This indicated the release of some paracrine factor from the mechanically stimulated cells. Application of ultrasonic vibration also induced an oscillation of intracellular Ca2+. The application of a uridine 5,-triphosphate (UTP), UTP, induced a transient increase in intracellular Ca2+ and the release of adenosine 5,-triphosphate (ATP) in cultured chondrocytes. A P2 receptor antagonist (suramin) and blockers of Cl, channels, niflumic acid and 4,4,-diisothiocyanostilbene-2,2,-disulfonic acid (DIDS), reduced the UTP-induced ATP release. The results indicated that Cl, channels were involved in the extracellular release of ATP following mechanical or P2Y receptor stimulation. Thus, ATP stimulation of P2Y receptors elicits an increase in intracellular Ca2+, triggering further release of ATP from adjacent cells, thereby expanding the Ca2+ wave in chondrocytes. Copyright © 2005 John Wiley & Sons, Ltd. [source] Structure-Based Synthetic Mimicry of Discontinuous Protein Binding Sites: Inhibitors of the Interaction of Mena EVH1 Domain with Proline-Rich LigandsCHEMBIOCHEM, Issue 8 2006Cornelia Hunke Dr. Abstract The Mena EVH1 domain, a protein-interaction module involved in actin-based cell motility, recognizes proline-rich ligand motifs, which are also present in the sequence of the surface protein ActA of Listeria monocytogenes. The interaction of ActA with host Mena EVH1 enables the bacterium to actively recruit host actin in order to spread into neighboring cells. Based on the crystal structure of Mena EVH1 in complex with a polyproline peptide ligand, we have generated a range of assembled peptides presenting the Mena EVH1 fragments that make up its discontinuous binding site for proline-rich ligands. Some of these peptides were found to inhibit the interaction of Mena EVH1 with the ligand pGolemi. One of them was further characterized at the level of individual amino acid residues; this yielded information on the contribution of individual positions of the peptides to the interaction with the ligand and identified sites for future structure optimization. [source] Oligomer-to-Polymer Transition in Short Ethylene Glycol Chains Connected to Mobile Hydrophobic AnchorsCHEMPHYSCHEM, Issue 1 2005Motomu Tanaka Dr. Abstract We studied the structure of short ethylene glycol (EG) chains with N repeating units (EGN, N=3, 6, 9, 12, and 15) connected to hydrophobic dihexadecyl chains by means of a combination of differential scanning calorimetry (DSC) and small- and wide-angle X-ray scattering (SAXS/WAXS). These synthetic amphiphiles dispersed in water form planar lamellar stacks and hexagonal cylinders confining the EG chains to restricted geometries. Owing to the self-assembly of the anchoring points, the lateral density of EG chains in planar lamella can be quantitatively controlled. Furthermore, the chain-melting phase transition of the anchors enables us to "switch" the intermolecular distance reversibly. SAXS/WAXS results suggest that the shorter EG chains (N=3, 6, and 9) assume a helical conformation in stacks of planar lamella. When the EG chains are further elongated (N=12 and 15), the lamellar periodicities cannot be explained by a linear extrapolation of shorter oligomers, but can be interpreted well as polymer brushes following the scaling theorem. Such rich phase behaviors of EGN molecules can be used as a simple model of oligo/poly-saccharide chains on cell surfaces, which act not only as flexible repellers between neighboring cells but also as stable spacers for functional ligands. [source] | ||||||||||||||||