scholarly journals Redistribution of surface macromolecules in dissociated epithelial cells.

1976 ◽  
Vol 71 (3) ◽  
pp. 907-920 ◽  
Author(s):  
M Pisam ◽  
P Ripoche
Keyword(s):  
Epithelial Cells ◽  
Cell Surface ◽  
Tight Junctions ◽  
Periodic Acid ◽  
Basal Membrane ◽  
Apical Surface ◽  
High Resolution Autoradiography ◽  
Dissociated Cells ◽  
The One ◽  
Surface Specialization

A number of ultrastructural and cytochemical techniques were used to study intact epithelial cells lining the frog urinary bladder: high resolution autoradiography after administration of [3H]glucosamine or [3H]fucose; 125I iodination of external protein; concanavalin A-peroxidase, periodic acid-chromic acid silver methenamine; and colloidal thorium. Results indicate that the material (probably glycoprotein) coating the apical surface differs from that which lines the lateral and basal surfaces. After dissociation and isolation of the epithelial cells, the material previously confined to the apical surface invaded progressively the opened "tight junctions" (about 5 min), then the lateral membranes (about 40 min), and finally the basal membrane (about 80 min): at that time, the whole cell surface was entirely enveloped by the apical material. Since, on the one hand, the reacting material was confined to the apical surface when the tight junctions were closed (in intact epithelial cells) and since, on the other hand, the apical material was sliding down the laterobasal membranes when the tight junctions were opened (in dissociated cells), it may be concluded that tight junctions contribute to maintain the cell surface specialization in epithelia.

Adhesion of uric acid crystals to the surface of renal epithelial cells

2000 ◽  
Vol 278 (6) ◽  
pp. F989-F998 ◽  
Author(s):  
Rima M. Koka ◽  
Erick Huang ◽  
John C. Lieske
Keyword(s):  
Uric Acid ◽  
Epithelial Cells ◽  
Cell Surface ◽  
Hydrophobic Interactions ◽  
Calcium Phosphates ◽  
Apical Cell ◽  
Apical Surface ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Renal Tubular Cells

Adhesion of microcrystals that nucleate in tubular fluid to the apical surface of renal tubular cells could be a critical step in the formation of kidney stones, 12% of which contain uric acid (UA) either alone or admixed with calcium oxalates or calcium phosphates. UA crystals bind rapidly to monolayer cultures of monkey kidney epithelial cells (BSC-1 line), used to model the surface of the nephron, in a concentration-dependent manner. The urinary glycoproteins osteopontin, nephrocalcin, and Tamm-Horsfall glycoprotein had no effect on binding of UA crystals to the cell surface, whereas other polyanions including specific glycosaminoglycans blocked UA crystal adhesion. Specific polycations also inhibited adhesion of UA crystals and appeared to exert their inhibitory effect by coating cells. However, removal of anionic cell surface molecules with neuraminidase, heparitinase I, or chondroitinase ABC each increased UA crystal binding, and sialic acid-binding lectins had no effect. These observations suggest that hydrogen bonding and hydrophobic interactions play a major role in adhesion of electrostatically neutral UA crystals to renal cells, unlike the interaction of calcium-containing crystals with negatively charged molecules on the apical cell surface via ionic forces. After adhesion to the plasma membrane, subsequent cellular events could contribute to UA crystal retention in the kidney and the development of UA or mixed calcium and UA calculi.

scholarly journals GAMBARAN MIKROSKOPIK GINJAL TIKUS WISTAR (RATTUS NORVEGICUS) SETELAH DIINDUKSI DENGAN GENTAMISIN

2013 ◽  
Vol 4 (3) ◽  
Author(s):  
Poppy M Lintong ◽  
Carla F Kairupan ◽  
Priska L N Sondakh
Keyword(s):  
Body Weight ◽  
Epithelial Cells ◽  
Wistar Rats ◽  
Periodic Acid ◽  
Basal Membrane ◽  
Tubular Epithelial Cells ◽  
Periodic Acid Schiff ◽  
Group Iii ◽  
Group I ◽  
Group Ii

Abstract: Gentamycin, a frequently used aminoglycoside antibiotics, has a nephrotoxic effect to human beings and animals. The purpose of this research was to find out the microscopic changes of wistar rat kidneys after gentamycin induction. This was an experimental study, using five adult wistar rats, divided into three groups. Group I was the control group; group II consisted of two rats, injected with gentamycin 0,3 ml/day (dose of 60 mg/kg body weight/day) intraperitoneally for seven days; and group III consisted of two rats, injected with gentamycin 0,3 ml/day intraperitoneally for 10 days. Group I and II were terminated at day-8, and group III at day-11. Their kidneys were processed for microscopic slides, stained with hematoxylin eosin and Periodic Acid Schiff. In microscopic evaluation, group II and III showed oedema, necrosis, apoptosis, and basal membrane destruction of tubular epithelial cells. Group III also showed fat vacuoles in these epithelial cells (macrovesicular fatty changes). Conclusion: wistar rats injected with gentamycin 60 mg/kg body weight/day for 7 and 10 days showed oedema, necrosis, apoptosis, and basal membrane destruction of tubular epithelial cells; and macrovesicular fatty changes after 10 days of gentamycin.Key words: gentamycin, necrosis tubular epithelial cells, fatty changesAbstrak: Gentamisin termasuk antibiotik golongan aminoglikosida berspektrum luas yang bersifat nefrotoksik terhadap manusia dan hewan. Tujuan penelitian ini untuk melihat perubahan mikroskopik struktur ginjal tikus Wistar setelah diberikan gentamisin. Metode penelitian eksperimental dengan menggunakan lima ekor tikus Wistar dewasa yang dibagi atas tiga kelompok. Kelompok I tanpa perlakuan; kelompok II terdiri dari dua ekor tikus perlakuan yang diinjeksi dengan gentamisin 0,3 ml/hari (dosis 60 mg/kgBB/hari) secara intraperitonial selama tujuh hari; dan kelompok III terdiri dari dua ekor tikus perlakuan yang diinjeksi dengan gentamisin 0,3 ml/hari secara intraperitonial selama 10 hari. Tikus Wistar kelompok I dan II diteminasi hari ke-8, sedangkan kelompok III diterminasi hari ke-11. Ginjal tikus kelompok I -III kemudian dibuat preparat histopatologik dengan pengecatan rutin hematoksilin eosin dan Periodic Acid Schiff (PAS). Hasil penelitian menunjukkan tikus Wistar perlakuan yang diberikan gentamisin 0,3 ml/hari selama 7 sampai 10 hari secara mikroskopik memperlihatkan pembengkakan, nekrosis, apoptosis, dan destruksi membrana basalis sel epitel tubulus; dan pada hari ke-10 terlihat vakuol-vakuol lemak pada sel epitel sehingga inti terdesak ke tepi (perlemakan makrovesikuler). Simpulan: pemberian gentamisin pada tikus Wistar dengan dosis 60 mg/kg BB/hari selama 7-10 hari menunjukkan pembengkakan, nekrosis, apoptosis sel epitel tubulus, dan membrana basalis tubulus rusak; dan setelah hari ke-10 juga terlihat perlemakan makrovesikuler.Kata kunci: gentamisin, nekrosis sel epitel tubulus, perlemakan makrovesikuler

Epithelial sorting of a glycosylphosphatidylinositol-anchored bacterial protein expressed in polarized renal MDCK and intestinal Caco-2 cells

1995 ◽  
Vol 108 (1) ◽  
pp. 369-377 ◽  
Author(s):  
K.L. Soole ◽  
M.A. Jepson ◽  
G.P. Hazlewood ◽  
H.J. Gilbert ◽  
B.H. Hirst
Keyword(s):  
Epithelial Cells ◽  
Cell Surface ◽  
Intestinal Epithelial Cells ◽  
Mdck Cells ◽  
Basolateral Membrane ◽  
Apical Cell ◽  
Apical Surface ◽  
Intestinal Epithelial ◽  
Gpi Anchor ◽  
Sorting Signal

To evaluate whether a glycosylphosphatidylinositol (GPI) anchor can function as a protein sorting signal in polarized intestinal epithelial cells, the GPI-attachment sequence from Thy-1 was fused to bacterial endoglucanase E' (EGE') from Clostridium thermocellum and polarity of secretion of the chimeric EGE'-GPI protein was evaluated. The chimeric EGE'-GPI protein was shown to be associated with a GPI anchor by TX-114 phase-partitioning and susceptibility to phosphoinositol-specific phospholipase C. In polarized MDCK cells, EGE' was localized almost exclusively to the apical cell surface, while in polarized intestinal Caco-2 cells, although 80% of the extracellular form of the enzyme was routed through the apical membrane over a 24 hour period, EGE' was also detected at the basolateral membrane. Rates of delivery of EGE'-GPI to the two membrane domains in Caco-2 cells, as determined with a biotinylation protocol, revealed apical delivery was approximately 2.5 times that of basolateral. EGE' delivered to the basolateral cell surface was transcytosed to the apical surface. These data indicate that a GPI anchor does represent a dominant apical sorting signal in intestinal epithelial cells. However, the mis-sorting of a proportion of EGE'GPI to the basolateral surface of Caco-2 cells provides an explanation for additional sorting signals in the ectodomain of some endogenous GPI-anchored proteins.

Adhesion of hydroxyapatite crystals to anionic sites on the surface of renal epithelial cells

1997 ◽  
Vol 273 (2) ◽  
pp. F224-F233 ◽  
Author(s):  
J. C. Lieske ◽  
R. Norris ◽  
F. G. Toback
Keyword(s):  
Epithelial Cells ◽  
Cell Surface ◽  
Alcian Blue ◽  
Crystal Surface ◽  
Cetylpyridinium Chloride ◽  
Apical Surface ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Subsequent Formation ◽  
Renal Tubular Cells

Adhesion of microcrystals that nucleate in tubular fluid to the apical surface of renal tubular cells could be a critical step in the formation of kidney stones, 20% of which contain hydroxyapatite (HA). HA crystals bound rapidly to monolayer cultures of monkey kidney epithelial cells (BSC-1 line), used to model the surface of the nephron, in a concentration-dependent manner. Adhesion was blocked by diverse polyanions including heparin, pentosan polysulfate, polyaspartate, and polyglutamate, as well as many found in tubular fluid such as chondroitin sulfates A and B, heparan sulfate, citrate, nephrocalcin, and osteopontin. The polycations cetylpyridinium chloride and cationized ferritin, as well as the cationic dyes alcian blue, polyethylenimine, and brilliant blue R, also inhibited adhesion of HA crystals, as did specific lectins including Triticum vulgaris (wheat germ agglutinin). Anions that inhibited adhesion of crystals appeared to act on the crystal surface, whereas cations and lectins exerted their effect on the cell. Treatment of cells with neuraminidase inhibited binding of crystals, suggesting that anionic cell surface sialic acid residues function as HA crystal receptor sites that can be blocked by specific cations or lectins. Adherence of HA crystals to cells of another renal line (MDCK) and, to 3T3 fibroblasts was also inhibited by heparin, polyaspartate, alcian blue, and T vulgaris lectin, suggesting that these crystals bind to analogous molecules on the surface of different types of cells. These results suggests that the structure, quantity, and/or function of soluble anions in tubular fluid, as well as those anchored to the cell surface, could be critical determinants of HA crystal retention in the nephron and the subsequent formation of a renal stone.

scholarly journals Trafficking of Shigella Lipopolysaccharide in Polarized Intestinal Epithelial Cells

1999 ◽  
Vol 145 (4) ◽  
pp. 689-698 ◽  
Author(s):  
Wandy L. Beatty ◽  
Stéphane Méresse ◽  
Pierre Gounon ◽  
Jean Davoust ◽  
Joëlle Mounier ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Tight Junctions ◽  
Apical Membrane ◽  
Intestinal Epithelial Cells ◽  
Fluid Phase ◽  
Apical Surface ◽  
Intestinal Epithelial ◽  
Electron Microscopic ◽  
Basolateral Side ◽  
Endocytic Compartments

Bacterial lipopolysaccharide (LPS) at the apical surface of polarized intestinal epithelial cells was previously shown to be transported from the apical to the basolateral pole of the epithelium (Beatty, W.L., and P.J. Sansonetti. 1997. Infect. Immun. 65:4395–4404). The present study was designed to elucidate the transcytotic pathway of LPS and to characterize the endocytic compartments involved in this process. Confocal and electron microscopic analyses revealed that LPS internalized at the apical surface became rapidly distributed within endosomal compartments accessible to basolaterally internalized transferrin. This compartment largely excluded fluid-phase markers added at either pole. Access to the basolateral side of the epithelium subsequent to trafficking to basolateral endosomes occurred via exocytosis into the paracellular space beneath the intercellular tight junctions. LPS appeared to exploit other endocytic routes with much of the internalized LPS recycled to the original apical membrane. In addition, analysis of LPS in association with markers of the endocytic network revealed that some LPS was sent to late endosomal and lysosomal compartments.

scholarly journals Characterization of epithelial membrane antigen expression in human mammary epithelium by ultrastructural immunoperoxidase cytochemistry.

1986 ◽  
Vol 34 (6) ◽  
pp. 801-809 ◽  
Author(s):  
O W Petersen ◽  
B van Deurs
Keyword(s):  
Epithelial Cells ◽  
Cell Surface ◽  
Milk Fat ◽  
Breast Biopsy ◽  
Intracellular Localization ◽  
Surface Membrane ◽  
Carcinoma Cells ◽  
Annulate Lamellae ◽  
Apical Surface ◽  
Human Breast Carcinomas

Ultrastructural immunocytochemistry was used to analyze cell surface distribution and intracellular localization of milk fat globule membrane antigen (MFGM-A) in cryosections from human breast carcinomas and benign breast biopsy specimens. The specimens were fixed in formaldehyde and frozen. Cryostat sections were cut at 15 micron, incubated with mouse monoclonal antibody to MFGM-A, and then with a peroxidase-conjugated goat anti-mouse antibody. After glutaraldehyde fixation, the sections were incubated with diaminobenzidine-H2O2 and further processed for electron microscopy. MFGM-A was specific for epithelial cells. MFGM-A staining was strictly confined to the apical surface membrane of normal ductal epithelium, never involving basolateral membranes below the tight junctions. In normal epithelial cells, MFGM-A was readily detected in cisternae of the endoplasmic reticulum (ER), but only to a lesser extent in Golgi complexes and presumptive secretory vesicles. In carcinoma cells, surface staining for MFGM-A was either distributed in a non-polarized manner on the entire cell surface or else was totally absent. In some carcinoma cells without surface-associated MFGM-A, very pronounced intracellular MFGM-A staining was seen in the ER, in the nuclear envelope, and in annulate lamellae. The observations on MFGM-A expression were supported by studies on a cell culture model system.

scholarly journals Activity and inhibition of prostasin and matriptase on apical and basolateral surfaces of human airway epithelial cells

2012 ◽  
Vol 303 (2) ◽  
pp. L97-L106 ◽  
Author(s):  
Shilpa Nimishakavi ◽  
Marina Besprozvannaya ◽  
Wilfred W. Raymond ◽  
Charles S. Craik ◽  
Dieter C. Gruenert ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Epithelial Cells ◽  
Cell Surface ◽  
Selective Inhibition ◽  
Airway Epithelial Cells ◽  
Bronchial Epithelial Cells ◽  
Apical Surface ◽  
Airway Epithelial ◽  
Wild Type ◽  
Bronchial Epithelial

Prostasin is a membrane-anchored protease expressed in airway epithelium, where it stimulates salt and water uptake by cleaving the epithelial Na+ channel (ENaC). Prostasin is activated by another transmembrane tryptic protease, matriptase. Because ENaC-mediated dehydration contributes to cystic fibrosis (CF), prostasin and matriptase are potential therapeutic targets, but their catalytic competence on airway epithelial surfaces has been unclear. Seeking tools for exploring sites and modulation of activity, we used recombinant prostasin and matriptase to identify substrate t-butyloxycarbonyl-l-Gln-Ala-Arg-4-nitroanilide (QAR-4NA), which allowed direct assay of proteases in living cells. Comparisons of bronchial epithelial cells (CFBE41o−) with and without functioning cystic fibrosis transmembrane conductance regulator (CFTR) revealed similar levels of apical and basolateral aprotinin-inhibitable activity. Although recombinant matriptase was more active than prostasin in hydrolyzing QAR-4NA, cell surface activity resisted matriptase-selective inhibition, suggesting that prostasin dominates. Surface biotinylation revealed similar expression of matriptase and prostasin in epithelial cells expressing wild-type vs. ΔF508-mutated CFTR. However, the ratio of mature to inactive proprostasin suggested surface enrichment of active enzyme. Although small amounts of matriptase and prostasin were shed spontaneously, prostasin anchored to the cell surface by glycosylphosphatidylinositol was the major contributor to observed QAR-4NA-hydrolyzing activity. For example, the apical surface of wild-type CFBE41o− epithelial cells express 22% of total, extractable, aprotinin-inhibitable, QAR-4NA-hydrolyzing activity and 16% of prostasin immunoreactivity. In conclusion, prostasin is present, mature and active on the apical surface of wild-type and CF bronchial epithelial cells, where it can be targeted for inhibition via the airway lumen.

scholarly journals Prostasin regulates epithelial monolayer function: cell-specific Gpld1-mediated secretion and functional role for GPI anchor

2006 ◽  
Vol 291 (6) ◽  
pp. C1258-C1270 ◽  
Author(s):  
George M. Verghese ◽  
Michael F. Gutknecht ◽  
George H. Caughey
Keyword(s):  
Epithelial Cells ◽  
Cell Surface ◽  
Collecting Duct ◽  
Critical Role ◽  
Lung Epithelial Cells ◽  
Peptidase Activity ◽  
Apical Surface ◽  
Gpi Anchor ◽  
Epithelial Monolayer ◽  
Lung Epithelial

Prostasin, a trypsinlike serine peptidase, is highly expressed in prostate, kidney, and lung epithelia, where it is bound to the cell surface, secreted, or both. Prostasin activates the epithelial sodium channel (ENaC) and suppresses invasion of prostate and breast cancer cells. The studies reported here establish mechanisms of membrane anchoring and secretion in kidney and lung epithelial cells and demonstrate a critical role for prostasin in regulating epithelial monolayer function. We report that endogenous mouse prostasin is glycosylphosphatidylinositol (GPI) anchored to the cell surface and is constitutively secreted from the apical surface of kidney cortical collecting duct cells. Using site-directed mutagenesis, detergent phase separation, and RNA interference approaches, we show that prostasin secretion depends on GPI anchor cleavage by endogenous GPI-specific phospholipase D1 (Gpld1). Secretion of prostasin by kidney and lung epithelial cells, in contrast to prostate epithelium, does not depend on COOH-terminal processing at conserved Arg322. Using stably transfected M-1 cells expressing wild-type, catalytically inactive, or chimeric transmembrane (not GPI)-anchored prostasins we establish that prostasin regulates transepithelial resistance, current, and paracellular permeability by GPI anchor- and protease activity-dependent mechanisms. These studies demonstrate a novel role for prostasin in regulating epithelial monolayer resistance and permeability in kidney epithelial cells and, furthermore, show specifically that prostasin is a critical regulator of transepithelial ion transport in M-1 cells. These functions depend on the GPI anchor as well as the peptidase activity of prostasin. These studies suggest that cell-specific Gpld1- or peptidase-dependent pathways for prostasin secretion may control prostasin functions in a tissue-specific manner.

scholarly journals Sorting of newly synthesized galactosphingolipids to the two surface domains of epithelial cells.

1996 ◽  
Vol 132 (5) ◽  
pp. 813-821 ◽  
Author(s):  
P van der Bijl ◽  
M Lopes-Cardozo ◽  
G van Meer
Keyword(s):  
Fatty Acid ◽  
Epithelial Cells ◽  
Cell Surface ◽  
Mdck Cells ◽  
Apical Surface ◽  
High Concentration ◽  
Selective Transport ◽  
Endogenous Lipid ◽  
Surface Domains ◽  
Golgi Network

The high concentration of glycosphingolipids on the apical surface of epithelial cells may be generated by selective transport from their site of synthesis to the cell surface. Previously, we showed that canine kidney MDCK and human intestinal Caco-2 cells converted a ceramide carrying the short fluorescent fatty acid C6-NBD to glucosylceramide (GlcCer) and sphingomyelin (SM), and that GlcCer was preferentially transported to the apical surface as compared to SM. Here, we address the point that not all glycosphingolipid classes are apically enriched in epithelia. We show that a ceramide containing the 2-hydroxy fatty acid C6OH was preferentially converted by MDCK and Caco-2 cells to galactosylceramide (GalCer) and its derivatives galabiosylceramide (Ga2Cer) and sulfatide (SGalCer) as compared to SM and GlcCer--all endogenous lipid classes of these cells. Transport to the apical and basolateral cell surface was monitored by a BSA-depletion assay. In MDCK cells, GalCer reached the cell surface with two- to sixfold lower apical/basolateral polarity than GlcCer. Remarkably, in Caco-2 cells GalCer and GlcCer displayed the same apical/basolateral polarity, but it was sixfold lower for lipids with a C6OH chain than for C6-NBD lipids. Therefore, the sorting of a sphingolipid appears to depend on lipid structure and cell type. We propose that the different ratios of gluco- and galactosphingolipid synthesis in the various epithelial tissues govern lipid sorting in the membrane of the trans Golgi network by dictating the composition of the domains from where vesicles bud to the apical and basolateral cell surface.

Adhesion of calcium oxalate monohydrate crystals to renal epithelial cells is inhibited by specific anions

1995 ◽  
Vol 268 (4) ◽  
pp. F604-F612 ◽  
Author(s):  
J. C. Lieske ◽  
R. Leonard ◽  
F. G. Toback
Keyword(s):  
Epithelial Cells ◽  
Cell Surface ◽  
Calcium Oxalate ◽  
Chondroitin Sulfate ◽  
Kidney Stones ◽  
Calcium Oxalate Monohydrate ◽  
Apical Surface ◽  
Dependent Manner ◽  
Renal Epithelial Cells ◽  
Renal Tubular Cells

Adhesion of urinary crystals to the apical surface of renal tubular cells could be a critical step in the formation of kidney stones. The interaction between renal epithelial cells (BSC-1 line) and the most common crystal in kidney stones, calcium oxalate monohydrate (COM), was studied in a tissue culture model system. COM crystals bound to the cell surface within seconds in a concentration-dependent manner to a far greater extent than did brushite, another calcium-containing crystal found in urine. Adhesion of COM crystals to cells was blocked by the polyanion, heparin. Other glycosaminoglycans including chondroitin sulfate A or B, heparan sulfate, and hyaluronic acid, but not chondroitin sulfate C, prevented binding of COM crystals. Two nonsulfated polyanions, polyglutamic acid and polyaspartic acid, also blocked adherence of COM crystals. Three molecules found in urine, nephrocalcin, uropontin, and citrate, each inhibited binding of COM crystals, whereas Tamm-Horsfall glycoprotein (THP) did not. Prior exposure of crystals but not cells to inhibitory molecules blocked adhesion, suggesting that these agents exert their effect at the crystal surface. Inhibition of crystal binding followed a linear Langmuir adsorption isotherm for each inhibitor identified, suggesting that these molecules bind to a single class of sites on the crystal that are important for adhesion to the cell surface. Inhibition of crystal adhesion by heparin was rapidly overcome by the polycation protamine, suggesting that the glycosaminoglycan regulates cell-crystal interactions in a potentially reversible manner.(ABSTRACT TRUNCATED AT 250 WORDS)

Sign in / Sign up

Export Citation Format

Share Document