Modification of fibroblast surface amines alters receptor-mediated cell spreading on protein-coated substrata but not adsorptive endocytosis

1981 ◽  
Vol 49 (1) ◽  
pp. 283-297
Author(s):  
J.D. Aplin ◽  
R.C. Hughes
Keyword(s):  
Cell Attachment ◽  
Surface Membrane ◽  
Fluorescein Isothiocyanate ◽  
Adhesive Interaction ◽  
Cultured Fibroblasts ◽  
Amine Groups ◽  
Coated Surfaces ◽  
Amine Compounds ◽  
Membrane Components ◽  
Dose Dependent

Fluorescein isothiocyanate (FITC) and other anionic reagents specific for amine groups have previously been shown to inhibit the adhesion and spreading of cultured fibroblasts to fibronectin-coated surfaces (Butters, Devalia, Aplin & Hughes, 1980). Here it is demonstrated that a population of FITC-labelled cells can be separated using flow cytometry into fractions displaying greater and lesser adhesivity at lower and higher fluorescence, respectively, demonstrating that the inhibition is dose-dependent. Glass coverslips covalently derivatized with the lectins ricin and concanavalin A are used to show that the inhibition also occurs in lectinmediated cell adhesion as well as in adhesion to collagen coated with fibronectin and plastic coated with serum or antibody, suggesting that all of these responses share a common, FITC-sensitive component. Simple primary amine compounds inhibit adhesion to fibronectin, but specific inhibitors of transglutaminases do not affect the process. Transglutaminase activity of cell surfaces has been implicated in protein endocytosis and receptor recycling (Davies et al. 1980). FITC modification of cells appears to affect specifically adhesive interaction, since ricin cytotoxicity and infection of cells with influenza and Sendai viruses (phenomena thought to proceed by means of receptor-mediated endocytosis) are unaffected. Evidently, receptor-mediated cell attachment, spreading on protein-coated surfaces and protein endocytosis are functionally separate events requiring different cell-surface membrane components, even when the same protein (ricin) is used to trigger these 2 processes.

The organization of cell surface membrane domains

1989 ◽  
Vol 47 ◽  
pp. 804-805
Author(s):  
Michael Edidin
Keyword(s):  
Cell Surface ◽  
Membrane Lipids ◽  
Surface Membrane ◽  
Lateral Diffusion ◽  
Cell Types ◽  
Membrane Domains ◽  
Membrane Biology ◽  
Morphological Polarity ◽  
Discrete Domains ◽  
Membrane Components

Cell surface membranes are based on a fluid lipid bilayer and models of the membranes' organization have emphasised the possibilities for lateral motion of membrane lipids and proteins within the bilayer. Two recent trends in cell and membrane biology make us consider ways in which membrane organization works against its inherent fluidity, localizing both lipids and proteins into discrete domains. There is evidence for such domains, even in cells without obvious morphological polarity and organization [Table 1]. Cells that are morphologically polarised, for example epithelial cells, raise the issue of membrane domains in an accute form.The technique of fluorescence photobleaching and recovery, FPR, was developed to measure lateral diffusion of membrane components. It has also proven to be a powerful tool for the analysis of constraints to lateral mobility. FPR resolves several sorts of membrane domains, all on the micrometer scale, in several different cell types.

Gonococci-human polymorphonuclear leukocyte interactions: metabolic studies associated with attachment and ingestion

1980 ◽  
Vol 28 (3) ◽  
pp. 991-1000
Author(s):  
A G Krieger ◽  
N L Schiller ◽  
R B Roberts
Keyword(s):  
Oxidative Metabolism ◽  
Polymorphonuclear Leukocytes ◽  
Surface Membrane ◽  
Optimal Conditions ◽  
Human Polymorphonuclear Leukocyte ◽  
Respiratory Inhibitors ◽  
Phase Type ◽  
Human Polymorphonuclear Leukocytes ◽  
Dose Dependent

Utilizing monolayers of human polymorphonuclear leukocytes, optimal conditions for attachment and ingestion of Neisseria gonorrhoeae were determined. Both attachment and ingestion were optimal at 36 degrees C when a bacteria-leukocyte ratio of 100:1 was employed. After 30 min of incubation, log-phase viable type 2 gonococci were attached to 90% of leukocytes, whereas log-phase viable type 4 gonococci were ingested by 80 to 90% of cells. Respiratory inhibitors had no effect on attachment or ingestion, whereas glycolytic inhibitors blocked ingestion but did not affect attachment of gonocci to the leukocyte surface. Inhibition was dose dependent and partially reversible. The oxidative metabolism of leukocytes with gonococci attached or ingested was also examined. Attachment of log-phase type 2 gonococci stimulated a minimal increase in glucose oxidation and oxygen consumption by leukocytes in contrast to marked increases by leukocytes that had ingested viable type 4 or heat-killed typed 2 organisms. These results demonstrate that attachment of log-phase type 2 gonococci to the surface membrane does not stimulate significant leukocyte oxidative metabolism nor initiate the phagocytic process.

scholarly journals Application of piezoelectric cells printing on three-dimensional porous bioceramic scaffold for bone regeneration

2019 ◽  
Vol 5 (2) ◽  
pp. 22 ◽  
Author(s):  
Ming-You Shie ◽  
Hsin-Yuan Fang ◽  
Yen-Hong Lin ◽  
Alvin Kai-Xing Lee ◽  
Joyce Yu ◽  
...  
Keyword(s):  
Cell Attachment ◽  
Three Dimensional ◽  
Fluorescein Isothiocyanate ◽  
Cellular Adhesion ◽  
Additive Manufacture ◽  
Cell Printing ◽  
Ceramic Scaffold ◽  
Porous Bioceramic ◽  
Bone Structures ◽  
Extrusion Technology

In recent years, the additive manufacture was popularly used in tissue engineering, as the various technologies for this field of research can be used. The most common method is extrusion, which is commonly used in many bioprinting applications, such as skin. In this study, we combined the two printing techniques; first, we use the extrusion technology to form the ceramic scaffold. Then, the stem cells were printed directly on the surface of the ceramic scaffold through a piezoelectric nozzle. We also evaluated the effects of polydopamine (PDA)-coated ceramic scaffolds for cell attachment after printing on the surface of the scaffold. In addition, we used fluorescein isothiocyanate to simulate the cell adhered on the scaffold surface after ejected by a piezoelectric nozzle. Finally, the attachment, growth, and differentiation behaviors of stem cell after printing on calcium silicate/polycaprolactone (CS/PCL) and PDACS/PCL surfaces were also evaluated. The PDACS/PCL scaffold is more hydrophilic than the original CS/PCL scaffold that provided for better cellular adhesion and proliferation. Moreover, the cell printing technology using the piezoelectric nozzle, the different cells can be accurately printed on the surface of the scaffold that provided and analyzed more information of the interaction between different cells on the material. We believe that this method may serve as a useful and effective approach for the regeneration of defective complex hard tissues in deep bone structures.

scholarly journals An analysis of lectin-initiated cell agglutination in a series of CHO subclones which respond morphologically to growth in dibutyryl cyclic AMP.

1976 ◽  
Vol 70 (1) ◽  
pp. 204-216 ◽  
Author(s):  
J van Veen ◽  
R M Roberts ◽  
K D Noonan
Keyword(s):  
Surface Membrane ◽  
Con A ◽  
Cell Agglutination ◽  
Lectin Receptors ◽  
Lectin Receptor ◽  
Free Mobility ◽  
Receptor Sites ◽  
Protein And Rna Synthesis ◽  
A Cell ◽  
Membrane Components

We have investigated the molecular basis of the agglutinability of CHO subclones which respond differentially in terms of morphology and surface architecture in the presence of dB-cAMP in the medium. We have demonstrated that the agglutinability of these subclones with both wheat germ agglutinin (WGA) and concanavalin A (Con A) probably depends on the free lateral mobility of the lectin receptor sites in the plane of the membrane. The nonagglutinable surface architecture seems to depend on the presence in the membrane of a protease-labile peptide(s), which appears to be distinct from the lectin receptors, as well as on continuous protein and RNA synthesis. This dependence on continuous transcription and translation may be related to the maintenance of the protease-labile peptide(s) in such a state as to restrict mobility of the lectin receptors. The surface architecture defined as nonagglutinable also depends on the state of polymerization of the intracellular microtubules and microfilaments. It is suggested that these microskeletal elements serve to anchor the lectin receptors in such a manner as to restrict their mobility and thereby reduce the relative agglutinability of a cell line. We suggest that control of the free mobility of both the Con A and WGA receptor sites is dependent on two constraints, one applied by protease-labile ("surface") membrane components and the other by components of the intracellular microskeletal system.

Detection of α1 Integrin in Urine of Patients With Immunoglobulin A Nephropathy

2008 ◽  
Vol 56 (3) ◽  
pp. 581-586
Author(s):  
Jonathan Bank ◽  
Aharon Ben-David ◽  
Ram Doolman ◽  
Ben-Ami Sela ◽  
Ilan Bank
Keyword(s):  
Mesangial Cells ◽  
Kidney Diseases ◽  
Surface Membrane ◽  
Immunoglobulin A ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dependent Manner ◽  
Healthy Individuals ◽  
Immunoglobulin A Nephropathy ◽  
A Cell ◽  
Dose Dependent

BackgroundThe α1β1 integrin is a cell surface membrane heterodimer composed of noncovalently linked α1 and β1 polypeptides that is up-regulated on activated and proliferating mesangial cells.MethodsA double-sandwich enzyme-linked immunosorbent assay that detects α1 integrin in a specific and dose-dependent manner at concentrations greater than 150 ng/mL was used to evaluate whether intact α1 polypeptides are secreted in the urine samples of 29 patients with various kidney diseases and in those of 5 healthy individuals.Resultsα1 Integrin was detected in 8 of the 29 patients including 3 of 3 patients with biopsy-proven immunoglobulin A nephropathy and 3 of 3 clinically suspected but non-biopsy-proven immunoglobulin A nephropathy with evidence of active nephritis. No α1 integrins were found in samples of 5 healthy controls.Conclusionsα1 Integrin polypeptides can be detected in human urine, particularly in immunoglobulin A nephropathy. Further extensive studies are required to clarify the significance of secretion of α1 integrins in urine of patients with kidney disease.

Alterations in cell surface membrane components of adapting rat small intestinal epithelium

1978 ◽  
Vol 75 (6) ◽  
pp. 1066-1072 ◽  
Author(s):  
Hugh J. Freeman ◽  
Marilynn E. Etzler ◽  
Arthur B. Garrido ◽  
Young S. Kim
Keyword(s):  
Cell Surface ◽  
Intestinal Epithelium ◽  
Surface Membrane ◽  
Small Intestinal Epithelium ◽  
Cell Surface Membrane ◽  
Small Intestinal ◽  
Membrane Components

Alloxan action on glucose metabolism in cultured fibroblasts. I. Stimulation and inhibition of glucose utilization

1981 ◽  
Vol 240 (6) ◽  
pp. E640-E644
Author(s):  
F. Ishibashi ◽  
H. Hidaka ◽  
R. M. Fields ◽  
B. V. Howard ◽  
P. H. Bennett
Keyword(s):  
Glucose Metabolism ◽  
Glucose Utilization ◽  
Cell Permeability ◽  
Dual Effect ◽  
Cultured Fibroblasts ◽  
Cell Monolayers ◽  
Glucose Incorporation ◽  
Dose Dependent ◽  
Cell Glucose ◽  
Fibroblastic Cells

The influence of alloxan on mammalian cell glucose metabolism has been investigated using human diploid fibroblastic cells in culture. When cell monolayers were exposed to D-[14C]glucose, the presence of alloxan (0.31–1.87 mM) resulted initially in a dose-dependent enhancement of total cell glucose incorporation. This was observed within 2 min and declined by 6 min. After that time, alloxan inhibited glucose incorporation. When hexose transport was examined directly using glucose analogues, alloxan neither enhanced nor inhibited the uptake of 3-O-methylglucose or 2-deoxyglucose. Alloxan exerted no effect on cell permeability or cell viability. These results suggest that alloxan may directly influence cell glucose metabolism beyond the level of phosphorylation. The dual effect of alloxan on glucose incorporation may be related to the alloxan stimulation and subsequent inhibition of glucose-induced insulin release in pancreatic islets.

scholarly journals Receptors for cold-insoluble globulin (plasma fibronectin) on human monocytes.

1981 ◽  
Vol 153 (1) ◽  
pp. 42-60 ◽  
Author(s):  
M P Bevilacqua ◽  
D Amrani ◽  
M W Mosesson ◽  
C Bianco
Keyword(s):  
Cell Attachment ◽  
Divalent Cations ◽  
Binding Activity ◽  
Membrane Receptors ◽  
Human Monocytes ◽  
Plasma Fibronectin ◽  
Blood Monocytes ◽  
Cold Insoluble Globulin ◽  
Particle Ingestion ◽  
Coated Surfaces

This investigation focused on the role played by cold-insoluble globulin (CIg, plasma fibronectin) in monocyte function. Surface-bound CIg mediated a concentration-dependent of human blood monocytes to gelatin-coated surfaces. CIg also mediated the binding of gelatin-coated particles such as latex beads or tanned erythrocytes to surface-bound human monocytes. However, CIg did not mediate particle ingestion. Subfractionated CIg that was highly enriched in monomeric forms (zone II CIg, mol wt 190,000-235,000) was less effective than were fractions enriched in dimeric forms (zone I CIg, mol wt 450,000) in promoting monocyte attachment. Binding of CIg to a gelatin or plastic surface occurred in the absence of divalent cations, but monocyte attachment to CIg-coated surfaces required divalent cations, Mg++ being much more effective than Ca++. Cation-dependent cell attachment was reversible in that bound cells could be released by treatment with EDTA. Serum-mediated binding of monocytes to gelatin-coated plastic dishes was a result of its content of CIg because the binding activity was abolished by removal of CIg from serum, and could be restored by readdition of purified CIg. Treatment of monocytes with trypsin abolished subsequent cell attachment to CIg-gelatin surfaces or particles. Expression of certain other known monocyte membrane receptors (Fc and C3b) was markedly enhanced as a result of CIg-monocyte interaction. These several observations indicate that monocytes bear membrane receptors (termed receptor cold-insoluble globulin) for surface-bound CIg.

Multiple pathways for denaturation of horse plasma gelsolin

1996 ◽  
Vol 74 (1) ◽  
pp. 101-107 ◽  
Author(s):  
Edward K. Koepf ◽  
Leslie D. Burtnick
Keyword(s):  
Surface Charge ◽  
Charge Distribution ◽  
Fluorescein Isothiocyanate ◽  
Heat Denaturation ◽  
Thermally Induced ◽  
Denatured State ◽  
Plasma Gelsolin ◽  
Surface Charge Distribution ◽  
Horse Plasma ◽  
Amine Groups

Gelsolin purified from horse plasma carries a surface charge distribution that greatly influences how the protein unfolds, aggregates, or precipitates as a function of temperature or concentration of chemical denaturant. Modification of gelsolin with fluorescein isothiocyanate replaces positive charges on amine groups with bulky, negatively charged fluorescein moieties. This postpones thermally induced precipitation by about 10 °C [Koepf, E.K., and Burtnick, L.D. 1993. Eur. J. Biochem. 212: 713–718]. Interaction with cations such as Ca2+ or guanidinium+ also alters the surface charge on gelsolin. This affects the structure of the protein in solution, modifies the pathway for unfolding, and moderates the onset of precipitation induced by chemical denaturants or heat. Denaturation of gelsolin is not interpretable in terms of a simple two-state cooperative mechanism. The pathway to a denatured state and intermediate structures present along the way depend upon the agent used to unfold the protein.Key words: gelsolin; denaturation, chemical, thermal, circular dichroism.

scholarly journals Lubiprostone Induces Claudin-1 and Protects Intestinal Barrier Function

Pharmacology ◽  
2019 ◽  
Vol 105 (1-2) ◽  
pp. 102-108 ◽  
Author(s):  
Norio Nishii ◽  
Tadayuki Oshima ◽  
Min Li ◽  
Hirotsugu Eda ◽  
Kumiko Nakamura ◽  
...  
Keyword(s):  
Barrier Function ◽  
Intestinal Barrier ◽  
Fluorescein Isothiocyanate ◽  
Intestinal Barrier Function ◽  
Epithelial Barrier ◽  
Dependent Manner ◽  
Epithelial Permeability ◽  
Epithelial Barrier Function ◽  
Dose Dependent

Introduction: Lubiprostone, a chloride channel activator, is said to reduce epithelial permeability. However, whether lubiprostone has a direct effect on the epithelial barrier function and how it modulates the intestinal barrier function remain unknown. Therefore, the effects of lubiprostone on intestinal barrier function were evaluated in vitro. Methods: Caco-2 cells were used to assess the intestinal barrier function. To examine the expression of claudins, immunoblotting was performed with specific antibodies. The effects of lubiprostone on cytokines (IFNγ, IL-6, and IL-1β) and aspirin-induced epithelial barrier disruption were assessed by transepithelial electrical resistance (TEER) and fluorescein isothiocyanate (FITC) labeled-dextran permeability. Results: IFNγ, IL-6, IL-1β, and aspirin significantly decreased TEER and increased epithelial permeability. Lubiprostone significantly improved the IFNγ-induced decrease in TEER in a dose-dependent manner. Lubiprostone significantly reduced the IFNγ-induced increase in FITC labeled-dextran permeability. The changes induced by IL-6, IL-1β, and aspirin were not affected by lubiprostone. The expression of claudin-1, but not claudin-3, claudin-4, occludin, and ZO-1 was significantly increased by lubiprostone. Conclusion: Lubiprostone significantly improved the IFNγ-induced decrease in TEER and increase in FITC labeled-dextran permeability. Lubiprostone increased the expression of claudin-1, and this increase may be related to the effect of lubiprostone on the epithelial barrier function.

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