scholarly journals Initial attachment of baby hamster kidney cells to an epoxy substratum. Ultrastructural analysis.

1976 ◽  
Vol 70 (3) ◽  
pp. 707-713 ◽  
Author(s):  
F Grinnell ◽  
M Q Tobleman ◽  
C R Hackenbrock
Keyword(s):  
Plasma Membrane ◽  
Cell Surface ◽  
Close Contact ◽  
Absolute Amount ◽  
Kidney Cells ◽  
Initial Attachment ◽  
Anionic Sites ◽  
Large Clusters ◽  
Baby Hamster Kidney Cells ◽  
Surface Collapse

In the presence of serum-containing medium, BHK cells attached and spread during a 1-h period onto a 3-5 nm thick serum layer absorbed on the substratum surface. The closest approach of the plasma membrane to the serum layer was observed to be about 9nm, which was determined by tilting the sectioned cells in a goniometer holder. Bundles of microfilaments or other cytoplasmic specializations were not observed in association with the regions of close contact. However, in the space between the plasma membrane and the adsorbed serum layer, a diffusely stained material could be visualized after fixation/staining by the tannic acid-glutaraldehyde technique. This technique also permitted increased clarity of visualization of trilaminar appearance of the plasma membrane. The distribution and mobility of anionic sites on the surfaces of attached and spreading cells was determined by labeling with polycationic ferritin. We observed movement of polycationic ferritin into large clusters on the cell surface, collapse of cell surface microextensions, and endocytosis, all of which were similar to our previous findings utilizing cells in suspension. However, the absolute amount of ferritin bound to the upper cell surface was less than that previously observed when suspended cells were put under similar labeling conditions. Also, polycationic ferritin did not appear to penetrate between the lower cell surface and the substratum.

scholarly journals The distribution and mobility of anionic sites on the surfaces of baby hamster kidney cells.

1975 ◽  
Vol 66 (3) ◽  
pp. 470-479 ◽  
Author(s):  
F Grinnell ◽  
M Q Tobleman ◽  
C R Hackenbrock
Keyword(s):  
Cell Surface ◽  
Cell Body ◽  
Cytochalasin B ◽  
Kidney Cells ◽  
Baby Hamster Kidney ◽  
Anionic Sites ◽  
Visual Probe ◽  
Entire Cell ◽  
Initial Binding ◽  
Baby Hamster Kidney Cells

The distribution and mobility of anionic sites on the surfaces of baby hamster kidney cells were studied by utilizing the multivalent ligand, polycationic ferritin, as a visual probe. Our observations revealed that anionic sites are distributed over the entire cell surface, with the highest density of sites being located on cell surface microextensions. Following the initial binding of polycationic ferritin to the surface of unfixed cells, the ligand-bound anionic sites redistributed by migrating from the surface of microextensions to the surface of the cell body. In 20 min, this migration resulted in a total clearing of anionic sites from the surface of microextensions concomitant with the formation of patches of anionic sites on the surface of the cell body. Polycationic ferritin-induced migration and patch formation of anionic sites was not prevented by 2,4-dinitrophenol, N-ethylmaleimide, colchicine, or cytochalasin B. However, the ligand-induced redistribution of cell surface anionic sites was prevented by prefixation of cells with glutaraldehyde.

BHK21 fibroblast aggregation inhibited by glycopeptides from the cell surface

1976 ◽  
Vol 21 (1) ◽  
pp. 161-173
Author(s):  
M.G. Vicker
Keyword(s):  
Cell Surface ◽  
Binding Sites ◽  
Cell Aggregation ◽  
The Other ◽  
Galactose Oxidase ◽  
Kidney Cells ◽  
Exclusion Chromatography ◽  
Baby Hamster Kidney Cells ◽  
High Voltage Electrophoresis ◽  
Complementary Binding

Glycopeptides were removed by trypsinization from the surface of baby hamster kidney cells (line BHK21-C13), digested by pronase and separated into 2 fractions by exclusion chromatography. The addition of small amounts of either glycopeptide fraction to shaken suspensions of lightly trypsinzied cells inhibited their rapid aggregation, but one fraction was more active than the other and in higher concentrations it was able to inhibit aggregation completely. After this fraction was purified by high-voltage electrophoresis one subfraction also inhibited aggregation. The effect of the glycopeptides increased following their pretreatment with neuraminidase, but preincubation with periodiate or galactose oxidase destroyed all activity. Galactose oxidase also inhibited cell aggregation directly. Similar glycopeptides from virus-transformed BHK21 cells, oligosaccharides and intact and desialysed human urinary glycoproteins had comparatively little or no effect on BHK21 cell aggregation. The results suggest terminal beta-galactosides and possible alpha-galactosides, and to some extent a particular substructure of cell surface heteroglycans are necessary for their inhibitory activity. The parent, plasma membrane of glycoproteins might serve as adhesive binding sites in cell cohesion, but some evidence indicates cell surface sialyl- and galactosyltransferases may not ordinarily act as their complementary binding receptors.

scholarly journals Rings of membrane sterols surround the openings of vesicles and fenestrae, in capillary endothelium.

1983 ◽  
Vol 97 (5) ◽  
pp. 1592-1600 ◽  
Author(s):  
N Simionescu ◽  
F Lupu ◽  
M Simionescu
Keyword(s):  
Plasma Membrane ◽  
Cell Membrane ◽  
Cell Surface ◽  
Freeze Fracture ◽  
Short Exposure ◽  
Capillary Endothelium ◽  
Anionic Sites ◽  
Microvascular Endothelium ◽  
Coated Pits ◽  
Plasmalemmal Vesicles

We investigated the distribution of sterols in the cell membrane of microvascular endothelium (mouse pancreas, diaphragm, brain, heart, lung, kidney, thyroid, adrenal, and liver) with the polyene antibiotic filipin, which reportedly has binding specificity for free 3-beta-hydroxysterols. In some experiments, concomitantly, cell-surface anionic sites were detected with cationized ferritin. Vessels were perfused in situ with PBS, followed by light fixation and filipin administration for 10 to 60 min. Tissues were further processed for thin-section and freeze-fracture electron microscopy. Short exposure (10 min) to filipin-glutaraldehyde solution resulted in the initial appearance, on many areas, of rings of characteristic filipin-sterol complexes within the rim surrounding stomata of most plasmalemmal vesicles, transendothelial channels, and fenestrae. Such rings were absent from the rims of the large openings of the sinusoid endothelium (liver, adrenal), coated pits and phagocytic vacuoles. After longer exposure (30-60 min), filipin-sterol complexes labeled randomly the rest of plasma membrane (except for coated pits, and partially the interstrand areas of junctions), and also marked most plasmalemmal vesicles. These peristomal rings of sterols were displayed mostly on the P face, and, at their full development, consisted of 6-8 units around a vesicle stoma, and 10-12 units around a fenestra. At their level, the intramembranous particles and the cell surface anionic sites were virtually excluded. Peristomal rings of sterols were also detected on the plasma membrane of pericytes and smooth muscle cells of the microvascular wall, which otherwise were poorly labeled with filipin-sterol complexes as compared to endothelial plasmalemma. It is presumed that the peristomal rings of cholesterol may represent important contributors to the local transient stabilization of plasma membrane and to the phase separation between cell membrane and vesicle membrane at a certain stage of their fusion/fission process.

scholarly journals Cell-surface mucosubstances from trypsin disaggregation of normal and virus-transformed lines of baby-hamster kidney cells (Short Communication)

1973 ◽  
Vol 134 (4) ◽  
pp. 1123-1126 ◽  
Author(s):  
S. Megan Minnikin ◽  
Adrian Allen
Keyword(s):  
Cell Surface ◽  
Cell Line ◽  
Cell Lines ◽  
Short Communication ◽  
Polyoma Virus ◽  
Kidney Cells ◽  
Baby Hamster Kidney ◽  
Transformed Cell ◽  
Transformed Cell Line ◽  
Baby Hamster Kidney Cells

Cell disaggregation by trypsin solubilizes significantly less mucosubstance from the surface of polyoma-virus-transformed baby-hamster kidney cells than from the same non-transformed cell line. The mucosubstance, which consists of both acid mucopolysaccharides and mucoproteins, also differs qualitatively in the two cell lines.

scholarly journals Immunocytochemical study of the partition and distribution of Sindbis virus glycoproteins in freeze-fractured membranes of infected baby hamster kidney cells.

1985 ◽  
Vol 101 (4) ◽  
pp. 1300-1306 ◽  
Author(s):  
M R Torrisi ◽  
S Bonatti
Keyword(s):  
Plasma Membrane ◽  
Sindbis Virus ◽  
Plasma Membranes ◽  
Protein A ◽  
Kidney Cells ◽  
Baby Hamster Kidney ◽  
Immunocytochemical Study ◽  
Viral Glycoproteins ◽  
Infected Cells ◽  
Baby Hamster Kidney Cells

Sindbis virus-infected baby hamster kidney cells were analyzed by thin section fracture-label. Specific immunolabel with antiviral glycoprotein antibodies or with conventional lectin label (wheat germ agglutinin) were used in conjunction with colloidal gold-conjugated protein A or ovomucoid, respectively. In addition, intact infected cells were analyzed with both labeling procedures. Experiments with Sindbis infected-chick embryo fibroblast cells were carried out as controls. Viral transmembrane glycoproteins appeared present in freeze-fractured inner and outer nuclear membrane, endoplasmic reticulum, Golgi stacks and vesicles, and plasma membranes; a clear preferential partition with the exoplasmic faces of all intracellular membranes was observed. By contrast, at the plasma membrane level, Sindbis glycoproteins were found to partition preferentially with the protoplasmic face. It seems likely that this protoplasmic partition is related to the binding with the nucleocapsid that takes place during the budding of the virus. At the cell surface, viral glycoproteins always appeared clustered and were predominantly associated with budding figures: moreover, large portions of the plasma membrane were devoid of both glycoproteins and budding viruses.

Cell surface receptors for wheat germ agglutinin and limulin in baby hamster kidney cells and ricin resistant variants

Biochimie ◽  
1981 ◽  
Vol 63 (3) ◽  
pp. 169-175 ◽  
Author(s):  
Angèle Obrenovitch ◽  
Claude Sene ◽  
Annie Claude Roche ◽  
Michel Monsigny ◽  
Peter Visher ◽  
...  
Keyword(s):  
Cell Surface ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Cell Surface Receptors ◽  
Kidney Cells ◽  
Baby Hamster Kidney ◽  
Surface Receptors ◽  
Baby Hamster Kidney Cells

scholarly journals Molecular characterization of gp40, a mucin-type glycoprotein from the apical plasma membrane of Madin–Darby canine kidney cells (type I)

1997 ◽  
Vol 326 (1) ◽  
pp. 99-108 ◽  
Author(s):  
Gert ZIMMER ◽  
Friedrich LOTTSPEICH ◽  
Andrea MAISNER ◽  
Hans-Dieter KLENK ◽  
Georg HERRLER
Keyword(s):  
Plasma Membrane ◽  
Cell Surface ◽  
Apical Plasma Membrane ◽  
Type I ◽  
Kidney Cells ◽  
Madin Darby Canine Kidney ◽  
A Cell ◽  
Polarized Transport ◽  
Darby Canine Kidney ◽  
Canine Kidney

gp40 has been recently identified as a major apical cell-surface sialoglycoprotein of type-I Madin–Darby canine kidney cells, a cell line widely used for the study of polarized transport. The determination of two internal amino acid sequences of the purified glycoprotein by Edman degradation enabled us to isolate the cDNA encoding the 18.6 kDa protein backbone of gp40. Sequence analysis revealed that gp40 is a type-I membrane protein which has several characteristics in common with glycophorin A and other mucin-type glycoproteins. At least 14 serine/threonine residues were found to be used for O-glycosylation. No potential sites for N-glycosylation were detected. gp40 turned out to represent the canine homologue of a cell-surface antigen expressed by various epithelial and non-epithelial cells in rat and mouse. Potential O-glycosylation sites, transmembrane and cytoplasmic domains were found to be highly conserved in the three species. gp40 was detected in canine lung, intestine, kidney, brain and heart but not in liver and spleen. The subline II of Madin–Darby canine kidney cells was found not to express gp40. Stable expression of gp40 in transfected type-II cells revealed that gp40 is predominantly delivered to the apical plasma membrane. N-Glycans and a glycosylphosphatidylinositol anchor, both proposed apical targeting signals, are absent from gp40, indicating that other determinants are responsible for its polarized transport.

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