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At least 667 records · Page 37Linked to original sources
PubMed1984

Serial alterations in endothelial cell shape and pattern after intraocular surgery.

We serially examined the endothelial cellular pattern and thickness of the central cornea after intracapsular cataract extraction and penetrating keratoplasty. The endothelial cellular pattern was analyzed with a computer-assisted digitizer. We used the frequency of hexagonal cells as the index to describe the degree of hexagonality in the cellular pattern. In the intracapsular cataract extraction group (20 eyes), the frequency of hexagonal cells was 68% preoperatively. During the first four weeks this frequency decreased rapidly to a significantly lower level (52%). The corneal thickness was significantly increased within the first week (0.57 mm) and steadily decreased to preoperative values at four weeks (0.53 mm). During the period from four to 24 weeks, the frequency of hexagonal cells increased gradually to 64%. In the penetrating keratoplasty group (ten eyes), the frequency of hexagonal cells and the thickness at one month were 62% and 0.64 mm respectively. During the next six months the frequency of hexagonal cells decreased significantly to 53%, and the thickness decreased to 0.53 mm. There was a gradual increase in the frequency of hexagonal cells to 62% during the two years after penetrating keratoplasty.

Aged↗
PubMed2001

The Ras-like GTPase Gem is involved in cell shape remodelling and interacts with the novel kinesin-like protein KIF9.

Gem belongs to the Rad/Gem/Kir (RGK) subfamily of Ras-related GTPases, which also comprises Rem, Rem2 and Ges. The RGK family members Ges and Rem have been shown to produce endothelial cell sprouting and reorganization of the actin cytoskeleton upon overexpression. Here we show that high intracellular Gem levels promote profound changes in cell morphology and we investigate how this phenotype arises dynamically. We also show that this effect requires intact microtubules and microfilaments, and that Gem is associated with both cytoskeletal components. In order to investigate the mechanisms of Gem recruitment to the cytoskeleton, we performed a yeast two-hybrid screen and identified a novel kinesin-like protein, termed KIF9, as a new Gem interacting partner. We further show that Gem and KIF9 interact by co-immunoprecipitation. Furthermore, Gem and KIF9 display identical patterns of gene expression in different tissues and developmental stages. The Gem- KIF9 interaction reported here is the first molecular link between RGK family members and the microtubule cytoskeleton.

Actin Cytoskeleton↗
PubMed2000

TGF-alpha differentially regulates GFAP, vimentin, and nestin gene expression in U-373 MG glioblastoma cells: correlation with cell shape and motility.

To begin understanding the regulation and biological significance of changes in the expression of intermediate filament proteins in astrocytic tumors, we have recently shown that TGF-alpha alters the protein level of glial fibrillary acidic protein (GFAP), nestin, and vimentin in U-373 MG glioblastoma cells. Here, we have determined the molecular mechanisms regulating these changes. In addition, to evaluate the significance of these changes we have examined whether TGF-alpha affects various cellular properties related to differentiation. Our results show that, in U-373 MG cells treated with TGF-alpha, GFAP gene transcription, mRNA level, and specific protein synthesis decrease by approximately 50%. This suggests that, in U-373 MG cells, TGF-alpha down-regulates the expression of this marker of astrocytic differentiation at the transcriptional level, resulting in decreased GFAP mRNA level and specific protein synthesis. In contrast, TGF-alpha does not change vimentin gene transcription, but increases by about 50% the transcription of the gene for nestin, a marker for undifferentiated astrocytic precursors. This differential regulation of GFAP, nestin, and vimentin gene expression indicates that TGF-alpha induces further dedifferentiation of U-373 MG cells. This notion is also supported by our findings that TGF-alpha increases the motility of U-373 MG cells and induces a less stellate morphology.

Cell Differentiation↗
PubMed1976

Cell shape changes induced by cationic anesthetics.

The effects of local anesthetics on cultivated macrophages were studied in living preparations and recorded in still pictures and time-lapse cine-micrographs. Exposure to 12mM lidocaine or 1.5 mM tetracaine resulted in rounding in 10-15 min. Rounding was characterized by cell contraction, marked increase in retraction fibrils, withdrawal of cell processes, and, in late stages, pulsation-like activity and zeiosis. Cells showed appreciable membrane activity as they rounded. Respreading was complete within 15 min of perfusion in drug-free medium and entailed a marked increase in surface motility over control periods. As many as eight successive cycles of rounding and spreading were obtained with lidocaine without evidence of cell damage. The effects of anesthetics were similar to those observed with EDTA, but ethylene-glycol-bis(beta-aminoethylether)-N, N'-tetraacetic acid-Mg was ineffective. Rounding was also induced by benzocaine, an anesthetic nearly uncharged at pH 7.0. Quaternary (nondischargeable) compounds were of low activity, presumably because they are slow permeants. Lidocaine induced rounding at 10 degrees C and above but was less effective at 5 degrees C and ineffective at 0 degrees C. Rounding by the anesthetic was also obtained in media depleted or Na or enriched with 10 mM Ca or Mg. The latter finding, together with the failure of tetrodotoxin to induce rounding, suggests that the anesthetic effect is unrelated to inhibition of sodium conductance. It is possible that the drugs influence divalent ion fluxes or some component of the contractile cells' machinery, but a metabolic target of action cannot yet be excluded.

Anesthetics, Local↗
PubMed1994

Mediation of chemoattractant-induced changes in [Ca2+]i and cell shape, polarity, and locomotion by InsP3, DAG, and protein kinase C in newt eosinophils.

During chemotaxis large eosinophils from newts exhibit a gradient of [Ca2+]i from rear to front. The direction of the gradient changes on relocation of the chemoattractant source, suggesting that the Ca2+ signal may trigger the cytoskeletal reorganization required for cell reorientation during chemotaxis. The initial stimulatory effect of chemoattractant on [Ca2+]i and the opposite orientations of the intracellular Ca2+ gradient and the external stimulus gradient suggest that more than one chemoattractant-sensitive messenger pathway may be responsible for the generation of spatially graded Ca2+ signals. To identify these messengers, Ca2+ changes were measured in single live cells stimulated with spatially uniform chemoattractant. On stimulation spatially averaged [Ca2+]i increased rapidly from < or = 100 nM to > or = 400 nM and was accompanied by formation of lamellipods. Subsequently cells flattened, polarized and crawled, and [Ca2+]i fluctuated around a mean value of approximately 200 nM. The initial Ca2+ spike was insensitive acutely to removal of extracellular Ca2+ but was abolished by treatments expected to deplete internal Ca2+ stores and by blocking receptors for inositol-trisphosphate, indicating that it is produced by discharge of internal stores, at least some of which are sensitive to InsP3. Activators of protein kinase C (PKC) (diacyl glycerol and phorbol ester) induced flattening and lamellipod activity and suppressed the Ca2+ spike, while cells injected with PKC inhibitors (an inhibitory peptide and low concentrations of heparin-like compounds) produced an enhanced Ca2+ spike on stimulation. Although cell flattening and lamellipod activity were induced by chemoattractant when the normal Ca2+ response was blocked, cells failed to polarize and crawl, indicating that Ca2+ homeostasis is required for these processes. We conclude that InsP3 acting on Ca2+ stores and DAG acting via PKC regulate chemoattractant-induced changes in [Ca2+]i, which in turn control polarization and locomotion. We propose that differences in the spatial distributions of InsP3 and DAG resulting from their respective hydrophilic and lipophilic properties may change Ca2+ distribution in response to stimulus reorientation, enabling the cell to follow the stimulus.

Animals↗
PubMed1996

A differential role for cell shape in neutrophil tethering and rolling on endothelial selectins under flow.

We investigated the role of neutrophil microvilli in interactions with E-selectin and P-selectin in hydrodynamic shear flow by disruption with cytochalasin B, hypotonic swelling, and chilling. Cytochalasin B only marginally reduced microvilli numbers (from 30 +/- 6 to 16 +/- 6 per cell perimeter, p < 0.005) as shown by electron microscopy, completely disrupted tethering in shear flow to E-selectin and P-selectin, increased the strength of rolling adhesions on E-selectin and P-selectin, and increased cell deformability in shear flow with a likely increase in the area of cell:substrate contact. Hypoosmotic swelling markedly reduced microvilli number (to 6 +/- 5 per perimeter, p < 0.005), almost completely inhibited tethering on E- and P-selectin, and increased the strength of rolling adhesions on P-selectin but not on E-selectin. Chilling almost completely abolished microvilli (to 3 +/- 3 per perimeter, p < 0.005), but pseudopod-like structures were present, and had little effect on tethering in flow. Immunogold labeling of L-selectin, which is normally clustered on tips of microvilli, showed that in the absence of microvilli it remained in small clusters. Our studies show that alterations in cell morphology and viscoelasticity can have opposing effects on tethering and rolling, showing that they are independently regulatable. Furthermore, our results suggest that the association of molecules that mediate rolling with microvilli tips may be important not just to enhance presentation, but for other functions such as to promote resistance to extraction from the membrane or cooperative interactions among clustered receptors.

Cell Adhesion↗
PubMed1999

Some relationships between membrane phospholipid domains, conformational order, and cell shape in intact human erythrocytes.

A novel method developed in this laboratory [D.J. Moore et al., Biochemistry 35 (1996) 229-235; D.J. Moore et al., Biochemistry 36 (1997) 660-664] to study the conformational order and the propensity for domain formation of specific phospholipids in intact human erythrocytes is extended to two additional species. Acyl chain perdeuterated 1,2-dilauroylphosphatidylethanolamine (diC12PE-d46) was incorporated preferentially (in separate experiments) into the inner leaflet of stomatocytic erythrocytes and into the outer leaflet of echinocytic erythrocytes, while acyl chain perdeuterated 1,2-dipentadecanoylphosphatidylcholine (diC15PC-d58) was incorporated into the outer leaflet of echinocytic erythrocytes. The conformational order and phase behavior of the incorporated molecules were monitored through FT-IR studies of the temperature dependence of the CD2 stretching vibrations. For both diC12PE-d46 and diC15PC-d58, the gel-->liquid crystal phase transition persisted when these lipids were located in the outer leaflet of echinocytic cells, a result indicative of the persistence of phospholipid domains. In each case, the transition widths were broadened compared to the pure lipids, suggestive of either small domains or the presence of additional molecular components within the domains. The conformational order of diC12PE-d46 differed markedly depending on its location and the morphology of the cells. When located predominantly in the inner membrane of stomatocytes, the phase transition of this species was abolished and the conformational order compared with pure lipid vesicles at the same temperature was much lower. The current results along with our previous studies provide a sufficient experimental basis to deduce some general principles of phospholipid conformational order and organization in both normal and shape-altered erythrocytes.

Cell Size↗
PubMed1992

Cell shape changes during transition of basal keratinocytes to mature enucleate-cornified envelopes: modulation of terminal differentiation by fibronectin.

Normal human keratinocytes (NHK) in submerged culture were used to assess mechanisms associated with in vitro exposure to known stimulators (sodium butyrate; NaB) and inhibitors (fibronectin; FN) of NHK maturation. A multiparameter approach was used to define cell types generated under a range of growth conditions. Differentiation induced in response to NaB occurred through a series of morphologically distinct phenotypes and culminated in the formation of enucleate-cornified envelopes. Two-dimensional electrophoresis provided a limited database to evaluate global changes in cellular protein composition as a function of induced differentiation. Proteins were identified that characterized predominantly basal cell cultures, NaB-treated cells, and fully differentiated NHKs. Growth of NHKs on FN suppressed both spontaneous and NaB-directed maturation and inhibited maximal expression of protein changes associated with the differentiated state. Thus, the composition of the extracellular matrix can modulate (at both the morphologic and protein levels) the response of basal NHKs to a potent differentiation-inducing agent. Abrogation of NHK maturation by FN was not due to adverse effects on cellular metabolism, abortive differentiation, or altered timing of induced differentiation. FN appears to exert its suppressive effect by either maintaining an early stem cell phenotype which is poorly competent for terminal maturation or attenuating an as yet unknown aspect of the NaB-initiated differentiation cascade.

Actins↗
PubMed1986

Spectrin, red cell shape and deformability. II. The antagonistic action of spectrin and sialic acid residues in determining membrane curvature in genetic spectrin deficiency in mice.

In a companion paper, the shapes of spectrin deficient mouse erythrocytes were described; in contrast to previous assumptions, spherules with tethered microvesicles rather than true "spherocytes" were found. Thence, spectrin deficient mouse erythrocytes are endowed with an excess of surface area for the given volume but the membrane is assuming a highly positive curvature. Observations during and after the action of enzymes cleaving the red cell surface charge (Neuraminidase, Trypsin, Chymotrypsin) showed that the previously positive membrane curvature, as well as the tendency of the membrane to flow into fingerlike protrusions was completely abolished. The erythrocytes of the spectrin deficient, desialylated mouse erythrocytes assumed a variety of shapes, often discocytic or even stomatocytic, i.e. their membrane presented with negative curvature. However, while these desialylated membranes could be easily deformed (elongated) by shear flow they did not recoil elastically into any definitive configuration after removal of the deforming forces. It is concluded from these observations that spectrin (acting on the inner interface between membrane and cytoplasm) and sialic acid residues (acting on the outer interface between membrane and plasma) exert antagonizing effects on membrane curvature and membrane bending elasticity. Sialic acid residues, strongly charged and situated on the outer side of the cell, produce positive membrane curvature; this observation can most readily be explained by assuming that this mechanical effect is caused by repulsive coulombic forces expanding the outer half of the bilayer. To explain the effect of the spectrin-complex in counteracting positive or in producing negative membrane curvature, a similar expansive coulombic force acting between the highly charged residues has been postulated. Thence, a model for explaining the overall elastic behaviour of the normal mammalian red cell is developed which is based on the assumption of elastic interactions of proteinacous membrane components coupled to the lipid bilayer of the membrane.

Animals↗
PubMed1980

Fusiform erythrocytes resembling sickle cells in angora goats: observations on osmotic and mechanical fragilities and reversal of cell shape during anaemia.

Osmotic and mechanical fragilities of erythrocytes were determined for seven goats having 3.4-71 per cent fusiform erythrocytes. The osmotic fragility was related to the erythrocyte shape in that the osmotic resistance was considerably higher for bloods containing more than 26 per cent fusiform erythrocytes. A decrease in the proportion of fusiform erythrocytes in the same goats was related to an increase in the osmotic fragility. Anaemia was induced in two goats by removal of 200-400 ml of blood at three or four day intervals for eight weeks. Red cell values decreased by 28-43 per cent within three weeks, but further bleeding produced either no or less (0-21 per cent) reductions in these values. Slight reticulocytosis was seen during the anaemic phase and there was a concomitant increase in the mean corpuscular volume and mean corpuscular haemoglobin values. Reticulocytosis diminished before the start of recovery from anaemia and disappeared during the recovery phase. The most significant finding was the change in the erythrocyte morphology during production of and recovery from anaemia. The development of anaemia was associated with a gradual reduction in the proportion of fusiform erythrocytes or discoid cells and simultaneous increase in the proportion of erythrocytes exhibiting distinct poikilocytosis. Recovery from the anaemia was rapid (within five weeks), but reversal of the erythrocyte shape took several months. Severe blood loss anaemia in the goat is known to induce synthesis of haemoglobin C, and in these anaemic goats formation of a new haemoglobin, most likely haemoglobin C, was demonstrated by electrophoretic and column chromatographic analyses. It was concluded that the formation of haemoglobin C was responsible for the morphological changes in the erythrocytes.

Anemia↗
PubMed1997

A Bacillus subtilis gene encoding a protein similar to nucleotide sugar transferases influences cell shape and viability.

Bacillus subtilis gene ypfP, which is located at 196 degrees on the genetic map, shows similarity to both the monogalactosyldiacylglycerol synthase gene of Cucumis sativus, which encodes a galactosyltransferase, and the murG genes of B. subtilis, Escherichia coli, Haemophilus influenzae, and Synechocystis sp. strain PCC6803, which encode N-acetylglucosaminyltransferases involved in peptidoglycan biosynthesis. Cells containing a null mutation of ypfP are shorter and rounder than wild-type cells during growth in Luria-Bertani medium and glucose minimal medium. In addition, the mutant cells preferentially undergo lysis when grown on solid Luria-Bertani medium.

Acyltransferases↗