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.

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.

scholarly journals Interactions between actin filaments and between actin filaments and membranes in quick-frozen and deeply etched hair cells of the chick ear.

1982 ◽  
Vol 95 (1) ◽  
pp. 249-261 ◽  
Author(s):  
N Hirokawa ◽  
L G Tilney
Keyword(s):  
Hair Cells ◽  
Actin Filaments ◽  
Apical Cell ◽  
Apical Surface ◽  
Contractile Ring ◽  
Vestibular Organ ◽  
Apical Cell Membrane ◽  
Cuticular Plate ◽  
Quick Freezing ◽  
Two Populations

Replicas of the apical surface of hair cells of the inner ear (vestibular organ) were examined after quick freezing and rotary shadowing. With this technique we illustrate two previously undescribed ways in which the actin filaments in the stereocilia and in the cuticular plate are attached to the plasma membrane. First, in each stereocilium there are threadlike connectors running from the actin filament bundle to the limiting membrane. Second, many of the actin filaments in the cuticular plate are connected to the apical cell membrane by tiny branched connecting units like a "crow's foot." Where these "feet" contact the membrane there is a small swelling. These branched "feet" extend mainly from the ends of the actin filaments but some connect the lateral surfaces of the actin filaments as well. Actin filaments in the cuticular plate are also connected to each other by finer filaments, 3 nm in thickness and 74 +/- 14 nm in length. Interestingly, these 3-nm filaments (which measure 4 nm in replicas) connect actin filaments not only of the same polarity but of opposite polarities as documented by examining replicas of the cuticular plate which had been decorated with subfragment 1 (S1) of myosin. At the apicolateral margins of the cell we find two populations of actin filaments, one just beneath the tight junction as a network, the other at the level of the zonula adherens as a ring. The latter which is quite substantial is composed of actin filaments that run parallel to each other; adjacent filaments often show opposite polarities, as evidenced by S1 decoration. The filaments making up this ring are connected together by the 3-nm connectors. Because of the polarity of the filaments this ring may be a "contractile" ring; the implications of this is discussed.

scholarly journals Differential Infection of Polarized Epithelial Cell Lines by Sialic Acid-Dependent and Sialic Acid-Independent Rotavirus Strains

2001 ◽  
Vol 75 (23) ◽  
pp. 11834-11850 ◽  
Author(s):  
Max Ciarlet ◽  
Sue E. Crawford ◽  
Mary K. Estes
Keyword(s):  
Sialic Acid ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Lines ◽  
Apical Cell ◽  
Paracellular Permeability ◽  
Apical Surface ◽  
Apical Cell Membrane ◽  
Basolateral Side ◽  
Epithelial Cell Lines

ABSTRACT Infection of epithelial cells by some animal rotaviruses, but not human or most animal rotaviruses, requires the presence ofN-acetylneuraminic (sialic) acid (SA) on the cell surface for efficient infectivity. To further understand how rotaviruses enter susceptible cells, six different polarized epithelial cell lines, grown on permeable filter membrane supports containing 0.4-μm pores, were infected apically or basolaterally with SA-independent or SA-dependent rotaviruses. SA-independent rotaviruses applied apically or basolaterally were capable of efficiently infecting both sides of the epithelium of all six polarized cell lines tested, while SA-dependent rotaviruses only infected efficiently through the apical surface of five of the polarized cell lines tested. Regardless of the route of virus entry, SA-dependent and SA-independent rotaviruses were released almost exclusively from the apical domain of the plasma membrane of polarized cells before monolayer disruption or cell lysis. The transepithelial electrical resistance (TER) of cells decreased at the same time, irrespective of whether infection with SA-independent rotaviruses occurred apically or basolaterally. The TER of cells infected apically with SA-dependent rotaviruses decreased earlier than that of cells infected basolaterally. Rotavirus infection decreased TER before the appearance of cytopathic effect and cell death and resulted in an increase in the paracellular permeability to [3H]inulin as a function of loss of TER. The presence of SA residues on either the apical or basolateral side was determined using a Texas Red-conjugated lectin, wheat germ agglutinin (WGA), which binds SA residues. WGA bound exclusively to SA residues on the apical surface of the cells, confirming the requirement for SA residues on the apical cell membrane for efficient infectivity of SA-dependent rotaviruses. These results indicate that the rotavirus SA-independent cellular receptor is present on both sides of the epithelium, but SA-dependent and SA-independent rotavirus strains infect polarized epithelial cells by different mechanisms, which may be relevant for pathogenesis and selection of vaccine strains. Finally, rotavirus-induced alterations of the epithelial barrier and paracellular permeability suggest that common mechanisms of pathogenesis may exist between viral and bacterial pathogens of the intestinal tract.

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 Retargeting the Coxsackievirus and Adenovirus Receptor to the Apical Surface of Polarized Epithelial Cells Reveals the Glycocalyx as a Barrier to Adenovirus-Mediated Gene Transfer

2000 ◽  
Vol 74 (13) ◽  
pp. 6050-6057 ◽  
Author(s):  
Raymond J. Pickles ◽  
Jill A. Fahrner ◽  
JenniElizabeth M. Petrella ◽  
Richard C. Boucher ◽  
Jeffrey M. Bergelson
Keyword(s):  
Gene Transfer ◽  
Epithelial Cells ◽  
Basolateral Membrane ◽  
Apical Cell ◽  
Airway Epithelial Cells ◽  
Apical Surface ◽  
Apical Cell Membrane ◽  
Human Airway ◽  
Coxsackievirus And Adenovirus Receptor ◽  
Adenovirus Receptor

ABSTRACT Lumenal delivery of adenovirus vectors (AdV) results in inefficient gene transfer to human airway epithelium. The human coxsackievirus and adenovirus receptor (hCAR) was detected by immunofluorescence selectively at the basolateral surfaces of freshly excised human airway epithelial cells, suggesting that the absence of apical hCAR constitutes a barrier to adenovirus-mediated gene delivery in vivo. In transfected polarized Madin-Darby canine kidney cells, wild-type hCAR was expressed selectively at the basolateral membrane, whereas hCAR lacking the transmembrane and/or cytoplasmic domains was expressed on both the basolateral and apical membranes. Cells expressing apical hCAR still were not efficiently transduced by AdV applied to the apical surface. However, after the cells were treated with agents that remove components of the apical surface glycocalyx, AdV transduction occurred. These results indicate that adenovirus can infect via receptors located at the apical cell membrane but that the glycocalyx impedes interaction of AdV with apical receptors.

Effect of Ethanol on Intracellular Vesicular Transport from Golgi to the Apical Cell Membrane

1998 ◽  
Vol 22 (1) ◽  
pp. 167 ◽  
Author(s):  
Amalia Slomiany ◽  
Pawel Nowak ◽  
Elizabeth Piotrowski ◽  
Bronislaw L. Slomiany
Keyword(s):  
Cell Membrane ◽  
Vesicular Transport ◽  
Apical Cell ◽  
Apical Cell Membrane

Differential signaling and regulation of apical vs. basolateral EGFR in polarized epithelial cells

1998 ◽  
Vol 275 (6) ◽  
pp. C1419-C1428 ◽  
Author(s):  
Scott K. Kuwada ◽  
Kirk A. Lund ◽  
Xiu Fen Li ◽  
Peter Cliften ◽  
Kurt Amsler ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Epithelial Cells ◽  
Cell Membrane ◽  
Apical Cell ◽  
Biologically Active ◽  
Apical Surface ◽  
Apical Cell Membrane ◽  
Collagen Protein ◽  
Epidermal Growth ◽  
Mediate Cell

Overexpression of the epidermal growth factor receptors (EGFR) in polarized kidney epithelial cells caused them to appear in high numbers at both the basolateral and apical cell surfaces. We utilized these cells to look for differences in the regulation and signaling of apical vs. basolateral EGFR. Apical and basolateral EGFR were biologically active and mediated EGF-induced cell proliferation to similar degrees. Receptor downregulation and endocytosis were less efficient at the apical surface, resulting in prolonged EGF-induced tyrosine kinase activity at the apical cell membrane. Tyrosine phosphorylation of EGFR substrates known to mediate cell proliferation, Src-homologous and collagen protein (SHC), extracellularly regulated kinase 1 (ERK1), and ERK2 could be induced similarly by activation of apical or basolateral EGFR. Focal adhesion kinase was tyrosine phosphorylated more by basolateral than by apical EGFR; however, β-catenin was tyrosine phosphorylated to a much greater degree following the activation of mislocalized apical EGFR. Thus EGFR regulation and EGFR-mediated phosphorylation of certain substrates differ at the apical and basolateral cell membrane domains. This suggests that EGFR mislocalization could result in abnormal signal transduction and aberrant cell behavior.

scholarly journals Isotropic actomyosin dynamics promote organization of the apical cell cortex in epithelial cells

2014 ◽  
Vol 207 (1) ◽  
pp. 107-121 ◽  
Author(s):  
Christoph Klingner ◽  
Anoop V. Cherian ◽  
Johannes Fels ◽  
Philipp M. Diesinger ◽  
Roland Aufschnaiter ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Apical Membrane ◽  
Myosin Ii ◽  
Apical Cell ◽  
Cell Cortex ◽  
Apical Surface ◽  
Cortical Actin ◽  
Cellular Mechanics ◽  
Apical Cell Membrane ◽  
Spatially Correlated

Although cortical actin plays an important role in cellular mechanics and morphogenesis, there is surprisingly little information on cortex organization at the apical surface of cells. In this paper, we characterize organization and dynamics of microvilli (MV) and a previously unappreciated actomyosin network at the apical surface of Madin–Darby canine kidney cells. In contrast to short and static MV in confluent cells, the apical surfaces of nonconfluent epithelial cells (ECs) form highly dynamic protrusions, which are often oriented along the plane of the membrane. These dynamic MV exhibit complex and spatially correlated reorganization, which is dependent on myosin II activity. Surprisingly, myosin II is organized into an extensive network of filaments spanning the entire apical membrane in nonconfluent ECs. Dynamic MV, myosin filaments, and their associated actin filaments form an interconnected, prestressed network. Interestingly, this network regulates lateral mobility of apical membrane probes such as integrins or epidermal growth factor receptors, suggesting that coordinated actomyosin dynamics contributes to apical cell membrane organization.

Apical heterodimeric cystine and cationic amino acid transporter expressed in MDCK cells

2002 ◽  
Vol 283 (1) ◽  
pp. F181-F189 ◽  
Author(s):  
Christian Bauch ◽  
François Verrey
Keyword(s):  
Mdck Cell ◽  
Mdck Cells ◽  
Apical Cell ◽  
Surface Expression ◽  
Xenopus Laevis Oocytes ◽  
Apical Surface ◽  
Apical Cell Membrane ◽  
Cationic Amino Acid ◽  
Confocal Immunofluorescence Microscopy ◽  
Apical Side

The luminal uptake ofl-cystine and cationic amino acids by (re)absorptive epithelia, as found in the small intestine and the proximal kidney tubule, is mediated by the transport system b0,+, which is defective in cystinuria. Expression studies in Xenopus laevis oocytes and other nonepithelial cells as well as genetic studies on cystinuria patients have demonstrated that two gene products, the glycoprotein rBAT and the multitransmembrane-domain protein b0,+AT, are required for system b0,+function. To study the biosynthesis, surface expression, polarity, and function of this heterodimer in an epithelial context, we established stable Madin-Darby canine kidney (MDCK) cell lines expressing rBAT and/or b0,+AT. Confocal immunofluorescence microscopy shows that both subunits depend on each other for apical surface expression. Immunoprecipitation of biosynthetically labeled proteins indicates that b0,+AT is stable in the absence of rBAT, whereas rBAT is rapidly degraded in the absence of b0,+AT. When both are coexpressed, they associate covalently and rBAT becomes fully glycosylated and more stable. Functional experiments show that the expressed transport is of the high-affinity b0,+-type and is restricted to the apical side of the epithelia. In conclusion, coexpression experiments in MDCK cell epithelia strongly suggest that the intracellular association of rBAT and b0,+AT is required for the surface expression of either subunit, which together form a functional heterocomplex at the apical cell membrane.

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