scholarly journals Antiadhesive Role of Apical Decay-accelerating Factor (CD55) in Human Neutrophil Transmigration across Mucosal Epithelia

2003 ◽  
Vol 198 (7) ◽  
pp. 999-1010 ◽  
Author(s):  
Donald W. Lawrence ◽  
Walter J. Bruyninckx ◽  
Nancy A. Louis ◽  
Douglas M. Lublin ◽  
Gregory L. Stahl ◽  
...  
Keyword(s):  
Cell Surface ◽  
Plasma Membranes ◽  
Apical Cell ◽  
Neutrophil Migration ◽  
Surface Proteins ◽  
Epithelial Surface ◽  
Decay Accelerating Factor ◽  
Apical Cell Membrane ◽  
Epithelial Monolayers ◽  
Epithelial Cell Surface

Neutrophil migration across mucosal epithelium during inflammatory episodes involves the precise orchestration of a number a cell surface molecules and signaling pathways. After successful migration to the apical epithelial surface, apically localized epithelial proteins may serve to retain PMN at the lumenal surface. At present, identification of apical epithelial ligands and their PMN counter-receptors remain elusive. Therefore, to define the existence of apical epithelial cell surface proteins involved in PMN–epithelial interactions, we screened a panel of antibodies directed against epithelial plasma membranes. This strategy identified one antibody (OE-1) that both localized to the apical cell membrane and significantly inhibited PMN transmigration across epithelial monolayers. Microsequence analysis revealed that OE-1 recognized human decay-accelerating factor (DAF, CD55). DAF is a highly glycosylated, 70–80-kD, glycosyl-phosphatidyinositol–linked protein that functions predominantly as an inhibitor of autologous complement lysis. DAF suppression experiments using antisense oligonucleotides or RNA interference revealed that DAF may function as an antiadhesive molecule promoting the release of PMN from the lumenal surface after transmigration. Similarly, peptides corresponding to the antigen recognition domain of OE-1 resulted in accumulation of PMN on the apical epithelial surface. The elucidation of DAF as an apical epithelial ligand for PMN provides a target for novel anti-inflammatory therapies directed at quelling unwanted inflammatory episodes.

scholarly journals Glycoconjugates on corneal epithelial surface: effect of neuraminidase treatment.

1989 ◽  
Vol 37 (8) ◽  
pp. 1215-1224 ◽  
Author(s):  
L D Hazlett ◽  
P Mathieu
Keyword(s):  
Plasma Membranes ◽  
Surface Effect ◽  
Apical Cell ◽  
Apical Surface ◽  
Corneal Surface ◽  
Epithelial Surface ◽  
Neuraminidase Treatment ◽  
Apical Cell Membrane ◽  
Corneal Epithelial ◽  
Adult Mice

The purpose of this study was to develop a procedure to quantitatively examine corneal epithelial apical cell membrane-associated glycoconjugates. Saccharide moieties on young, mature, and aged corneal epithelial cells were detected and localized in corneas of immature and adult mice by using colloidal gold-labeled lectins and transmission electron microscopy (TEM). In general, dense binding to the corneal epithelial apical surface cell membranes with wheat germ agglutinin (WGA) was seen in the adult, whereas the immature cornea bound less WGA-gold. Neuraminidase digestion decreased binding of the conjugate on epithelial plasma membranes of young and mature cells in adult cornea. Lectin-gold binding was decreased in the immature cornea on mature and aged cells. WGA-gold binding after neuraminidase was elevated on young cells of immature and on aged cells of adult animals. No binding of peanut agglutinin (PNA) or horse gram agglutinin (DBA) to the corneal epithelial surface was seen in animals of either age. After neuraminidase digestion, PNA binding sites were exposed only on the adult corneal surface. These data suggest that a terminal trisaccharide sequence, sialic acid-galactose beta(1----3)-N-acetylgalactosamine, is present at the adult corneal surface but is absent or at undetectable levels at the corneal surface of the immature animal. These data may be of significance in light of the dissimilar pattern of P. aeruginosa recognition and binding to the immature vs adult corneal epithelium.

Delivery of newly synthesized Na(+)-K(+)-ATPase to the plasma membrane of A6 epithelia

1997 ◽  
Vol 272 (6) ◽  
pp. C1781-C1789 ◽  
Author(s):  
B. Coupaye-Gerard ◽  
J. B. Zuckerman ◽  
P. Duncan ◽  
A. Bortnik ◽  
D. I. Avery ◽  
...  
Keyword(s):  
Steady State ◽  
Cell Surface ◽  
Apical Cell ◽  
Surface Proteins ◽  
Beta Subunit ◽  
Alpha Subunit ◽  
Epithelial Cell Line ◽  
Steady State Distribution ◽  
State Distribution ◽  
Sorting Signals

Na(+)-K(+)-ATPase is localized to the basolateral cell surface of most epithelial cells. Conflicting results regarding the intracellular trafficking of Na(+)-K(+)-ATPase in Madin-Darby canine kidney cells have been reported, with delivery to both apical and basolateral membranes or exclusively to the basolateral cell surface. We examined the delivery and steady-state distribution of Na(+)-K(+)-ATPase in the amphibian epithelial cell line A6 using an antibody raised against Na(+)-K(+)-ATPase alpha-subunit and sulfo-N-hydroxysuccinimidobiotin to tag cell surface proteins. The steady-state distribution of the Na(+)-K(+)-ATPase was basolateral, as confirmed by immunocytochemistry. Delivery of newly synthesized Na(+)-K(+)-ATPase to the cell surface was examined using [35S]methionine and [35S]cysteine in a pulse-chase protocol. After a 20-min pulse, the alpha-subunit and core glycosylated beta-subunit were present at both apical and basolateral cell surfaces. The alpha-subunit and core glycosylated beta-subunit delivered to the apical cell surface were degraded within 2 h. Mature alpha/beta-heterodimer was found almost exclusively at the basolateral surface after a 1- to 24-h chase. These data suggest that immature Na(+)-K(+)-ATPase alpha-subunit and core glycosylated beta-subunits are not retained in the endoplasmic reticulum of A6 cells and apparently lack sorting signals. Mature Na(+)-K(+)-ATPase is targeted to the basolateral surface, suggesting that basolateral targeting of the protein is conformation dependent.

scholarly journals Vesicular transport of cationized ferritin by the epithelium of the rat choroid plexus.

1981 ◽  
Vol 89 (1) ◽  
pp. 131-139 ◽  
Author(s):  
B Van Deurs ◽  
F Von Bülow ◽  
M Møller
Keyword(s):  
Choroid Plexus ◽  
Vesicular Transport ◽  
Apical Cell ◽  
Multivesicular Bodies ◽  
Apical Surface ◽  
Cationized Ferritin ◽  
Epithelial Surface ◽  
Apical Cell Membrane ◽  
Choroidal Epithelium ◽  
Limited Degree

We have studied the transport of ferritin that was internalized by coated micropinocytic vesicles at the apical surface of the choroid plexus epithelium in situ. After ventriculocisternal perfusion of native ferritin (NF) or cationized ferritin (CF), three routes followed by the tracers are revealed: (a) to lysosomes, (b) to cisternal compartments, and (c) to the basolateral cell surface. (a) NF is micropinocytosed to a very limited degree and appears in a few lysosomal elements whereas CF is taken up in large amounts and can be followed, via endocytic vacuoles and light multivesicular bodies, to dark multivesicular bodies and dense bodies. (b) Occasionally, CF particles are found in cisterns that may represent GERL or trans-Golgi elements, whereas stacked Golgi cisterns never contain CF. (c) Transepithelial vesicular transport of CF is distinctly revealed. The intercellular spaces of the epithelium, below the apical tight junctions, contain numerous clusters of CF particles, often associated with surface-connected, coated vesicles. Vesicles in the process of exocytosis of CF are also present at the basal epithelial surface, whereas connective tissue elements below the epithelium are unlabeled. Our conclusion is that fluid and solutes removed from the cerebrospinal fluid by endocytosis either become sequestered in the lysosomal apparatus of the choroidal epithelium or are transported to the basolateral surface. However, our results do not indicate any significant recycling via Golgi complexes of internalized apical cell membrane.

MUCI, glycans and the cell-surface barrier to embryo implantation

2001 ◽  
Vol 29 (2) ◽  
pp. 153-156 ◽  
Author(s):  
J. D. Aplin ◽  
M. Meseguer ◽  
C. Simon ◽  
M. E. Ortíz ◽  
H. Croxatto ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Epithelial Cells ◽  
Cell Surface ◽  
Embryo Implantation ◽  
Human Embryos ◽  
Surface Barrier ◽  
Epithelial Surface ◽  
Epithelial Monolayers ◽  
Maternal Rejection ◽  
Normal Expression

As it approaches the maternal surface, the attaching embryo encounters the epithelial glycocalyx, which contains the mucin, MUC1. A high density of MUC1 at the cell surface can inhibit cell adhesion. This raises the possibility of the existence of a uterine barrier to implantation that might allow maternal rejection of poorer quality embryos. To investigate the mechanism of implantation, human embryos were incubated with endometrial epithelial monolayers. Hatched blastocysts were found to attach readily to the epithelial surface. MUC1 was lost from epithelial cells beneath and near to the attached embryo, while normal expression persisted in neighbouring cells.

scholarly journals Collagen binding proteins derived from the embryonic fibroblast cell surface recognize arginine-glycine-aspartic acid

1989 ◽  
Vol 9 (3) ◽  
pp. 329-340 ◽  
Author(s):  
Roy C. Ogle ◽  
Charles D. Little
Keyword(s):  
Cell Surface ◽  
Aspartic Acid ◽  
Plasma Membranes ◽  
Binding Proteins ◽  
Fibroblast Cell ◽  
Surface Proteins ◽  
Collagen Molecule ◽  
Collagen Binding ◽  
Arginine Glycine Aspartic Acid ◽  
Embryonic Fibroblast

Several cell surface proteins (Mr=120,000, 90,000, 63,000 and 47,000) apparently integral to embryonic fibroblast plasma membranes were extracted with detergent and isolated by collagen affinity chromatography. Certain of these proteins (Mr=120,000, 90,000 and 47,000) were specifically eluted from collagen affinity columns by synthetic peptides containing the amino acid sequence arginyl-glycyl-aspartic acid (RGD). These data show that a number of collagen binding proteins exist on the embryonic fibroblast cell surface. Some of the proteins may be collagen receptors binding to RGD sequences in the collagen molecule while at least one of the proteins (Mr=63,000) recognizes features other than RGD.

Neutral sphingomyelinase inhibitor scyphostatin prevents and ceramide mimics mechanotransduction in vascular endothelium

2004 ◽  
Vol 287 (3) ◽  
pp. H1344-H1352 ◽  
Author(s):  
Malgorzata Czarny ◽  
Jan E. Schnitzer
Keyword(s):  
Endothelial Cell ◽  
Cell Surface ◽  
Plasma Membranes ◽  
Fluid Shear Stress ◽  
Experimental Models ◽  
Surface Proteins ◽  
Rat Lung ◽  
Neutral Sphingomyelinase

Recently, we showed that neutral sphingomyelinase (N-SMase) is concentrated at the endothelial cell surface in caveolae and is activated to produce ceramide in an acute and transient manner by increase in flow rate and pressure in rat lung vasculature (Czarny M, Liu J, Oh P, and Schnitzer JE, J Biol Chem 278: 4424–4430, 2003). Here, we report further on our investigations of this new acute mechanotransduction pathway. We employed three experimental models to explore the role of N-SMase and ceramides in mechanosignaling: 1) a cell-free, in vitro model using isolated luminal plasma membranes of rat lung endothelium; 2) a fluid shear stress model using monolayers of intact bovine aorta endothelial cell in culture; and 3) an in situ model using controlled perfusion of the rat lung vasculature. Scyphostatin, which specifically inhibited N-SMase but not acid SMase activity, prevented mechanoactivation of N-SMase as well as downstream tyrosine and mitogen-activated protein kinases. Cell-permeable ceramide analogs ( N-acetylsphingosine, C2-ceramide, and N-hexanoylsphingosine, C6-ceramide) but not the inactive dihydroderivatives D2-ceramide and D6-ceramide ( N-acetylsphinganine and N-hexanoylsphinganine, respectively) mimic rapid mechano-induced tyrosine phosphorylation of cell surface proteins as well as mechanoactivation of Src-like kinases and the extracellular regulated kinase pathway. The responses common to ceramide and mechanical stress were inhibited by genistein, herbamycin A, and PP2, but not PP3, which suggests an obligate role of Src-like kinases in ceramide-mediated mechanotransduction. Ceramides also induced serine/threonine phosphorylation to activate the Akt/endothelial nitric oxide synthase pathway. Thus N-SMase at the plasma membrane in caveolae may be an upstream initiating mechanosensor, which acutely triggers mechanotransduction by generation of the lipid second messenger ceramide.

scholarly journals Neisseria gonorrhoeae Induces Focal Polymerization of Actin in Primary Human Urethral Epithelium

1998 ◽  
Vol 66 (7) ◽  
pp. 3416-3419 ◽  
Author(s):  
Peter C. Giardina ◽  
Richard Williams ◽  
David Lubaroff ◽  
Michael A. Apicella
Keyword(s):  
Epithelial Cell ◽  
Cell Surface ◽  
Cell Lines ◽  
Conjunctival Epithelium ◽  
Cytoskeletal Reorganization ◽  
Epithelial Surface ◽  
Immortalized Cell Lines ◽  
Epithelial Cell Surface ◽  
Immortalized Cell ◽  
Cytoskeletal Rearrangements

ABSTRACT The pathogenic Neisseria species induce cytoskeletal reorganization in immortalized cell lines. In Chang conjunctival epithelium and T84 intestinal epithelium, focal cytoskeletal rearrangements in which bacteria contacted the epithelial surface were observed. We show that actin footprints are induced in gonococcus-challenged primary urethral epithelium. Moreover, the microbes induced microvillus extension from the epithelial cell surface. Our results indicate that formation of actin footprints is not an artifact of commonly used immortalized cell lines.

scholarly journals A 39-kD plasma membrane protein (IP39) is an anchor for the unusual membrane skeleton of Euglena gracilis.

1990 ◽  
Vol 110 (4) ◽  
pp. 1077-1088 ◽  
Author(s):  
T K Rosiere ◽  
J A Marrs ◽  
G B Bouck
Keyword(s):  
Plasma Membrane ◽  
Membrane Protein ◽  
Cell Surface ◽  
Euglena Gracilis ◽  
Plasma Membranes ◽  
Membrane Skeleton ◽  
Surface Proteins ◽  
Plasma Membrane Protein ◽  
Octyl Glucoside ◽  
Skeletal Protein

The major integral plasma membrane protein (IP39) of Euglena gracilis was radiolabeled, peptide mapped, and dissected with proteases to identify cytoplasmic domains that bind and anchor proteins of the cell surface. When plasma membranes were radioiodinated and extracted with octyl glucoside, 98% of the extracted label was found in IP39 or the 68- and 110-kD oligomers of IP39. The octyl glucoside extracts were incubated with unlabeled cell surface proteins immobilized on nitrocellulose (overlays). Radiolabel from the membrane extract bound one (80 kD) of the two (80 and 86 kD) major membrane skeletal protein bands. Resolubilization of the bound label yielded a radiolabeled polypeptide identical in Mr to IP39. Intact plasma membranes were also digested with papain before or after radioiodination, thereby producing a cytoplasmically truncated IP39. The octyl glucoside extract of truncated IP39 no longer bound to the 80-kD membrane skeletal protein in the nitrocellulose overlays. EM of intact or trypsin digested plasma membranes incubated with membrane skeletal proteins under stringent conditions similar to those used in the nitrocellulose overlays revealed a partially reformed membrane skeletal layer. Little evidence of a membrane skeletal layer was found, however, when plasma membranes were predigested with papain before reassociation. A candidate 80-kD binding domain of IP39 has been tentatively identified as a peptide fragment that was present after trypsin digestion of plasma membranes, but was absent after papain digestion in two-dimensional peptide maps of IP39. Together, these data suggest that the unique peripheral membrane skeleton of Euglena binds to the plasma membrane through noncovalent interactions between the major 80-kD membrane skeletal protein and a small, papain sensitive cytoplasmic domain of IP39. Other (62, 51, and 25 kD) quantitatively minor peripheral proteins also interact with IP39 on the nitrocellulose overlays, and the possible significance of this binding is discussed.

Polarized insertion of an intracellular glycoprotein pool into the apical membrane of MDCK cells

1990 ◽  
Vol 258 (3) ◽  
pp. C390-C398 ◽  
Author(s):  
G. K. Ojakian ◽  
R. Schwimmer ◽  
R. E. Herz
Keyword(s):  
Cell Surface ◽  
Plasma Membranes ◽  
Apical Membrane ◽  
Mdck Cells ◽  
Apical Cell ◽  
Temperature Shift ◽  
Immunogold Electron Microscopy ◽  
Trypsin Treatment ◽  
Cytoplasmic Vesicles ◽  
Darby Canine Kidney

A monoclonal antibody that recognizes a 135-kDa glycoprotein (GP135) on the apical membrane of Madin-Darby canine kidney (MDCK) cells was used to identify and characterize an intracellular pool of GP135. Mild trypsin treatment at 4 degrees C removed approximately 95% of the GP135, and after warming to 37 degrees C, the reappearance of GP135 on the apical membrane was monitored by radioimmunoassay. Incorporation of GP135 into the apical cell surface after trypsin treatment consisted of two components, a rapidly inserted, cycloheximide-insensitive portion (defined as the GP135 pool), which leveled off within 1 h, followed by a slower insertion of newly synthesized GP135. Immunogold electron microscopy demonstrated that the GP135 pool was targeted in a polarized manner and was only detected on the apical membrane. Temperature shift and retrypsinization experiments provided evidence that the GP135 pool consisted of intracellular vesicles that could fuse with the plasma membrane. This was confirmed by immunofluorescence microscopy demonstrating that GP135 was localized within large cytoplasmic vesicles residing at varying distances from the apical cell surface. These data provide evidence for the presence of a regulated pathway in MDCK cells and support the possibility that the GP135 pool functions as an intracellular reserve which can exhibit polarized insertion into the plasma membranes similar to that described for other epithelial cells.

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