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 Caenorhabditis elegans chaperonin CCT/TRiC is required for actin and tubulin biogenesis and microvillus formation in intestinal epithelial cells

2014 ◽  
Vol 25 (20) ◽  
pp. 3095-3104 ◽  
Author(s):  
Keiko Saegusa ◽  
Miyuki Sato ◽  
Katsuya Sato ◽  
Junko Nakajima-Shimada ◽  
Akihiro Harada ◽  
...  
Keyword(s):  
Caenorhabditis Elegans ◽  
Epithelial Cells ◽  
Apical Membrane ◽  
Intestinal Epithelial Cells ◽  
Intestinal Cells ◽  
Intermediate Filament Protein ◽  
Apical Surface ◽  
Intestinal Epithelial ◽  
Microtubule Network

Intestinal epithelial cells have unique apical membrane structures, known as microvilli, that contain bundles of actin microfilaments. In this study, we report that Caenorhabditis elegans cytosolic chaperonin containing TCP-1 (CCT) is essential for proper formation of microvilli in intestinal cells. In intestinal cells of cct-5(RNAi) animals, a substantial amount of actin is lost from the apical area, forming large aggregates in the cytoplasm, and the apical membrane is deformed into abnormal, bubble-like structures. The length of the intestinal microvilli is decreased in these animals. However, the overall actin protein levels remain relatively unchanged when CCT is depleted. We also found that CCT depletion causes a reduction in the tubulin levels and disorganization of the microtubule network. In contrast, the stability and localization of intermediate filament protein IFB-2, which forms a dense filamentous network underneath the apical surface, appears to be superficially normal in CCT-deficient cells, suggesting substrate specificity of CCT in the folding of filamentous cytoskeletons in vivo. Our findings demonstrate physiological functions of CCT in epithelial cell morphogenesis using whole animals.

The effect of berberine in vitro on tight junctions in human Caco-2 intestinal epithelial cells

Fitoterapia ◽  
2009 ◽  
Vol 80 (4) ◽  
pp. 241-248 ◽  
Author(s):  
Lili Gu ◽  
Ning Li ◽  
Qiurong Li ◽  
Qiang Zhang ◽  
Chengyang Wang ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Tight Junctions ◽  
Intestinal Epithelial Cells ◽  
Intestinal Epithelial

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.

scholarly journals Cell-adhesion molecule uvomorulin is localized in the intermediate junctions of adult intestinal epithelial cells.

1985 ◽  
Vol 100 (1) ◽  
pp. 327-332 ◽  
Author(s):  
K Boller ◽  
D Vestweber ◽  
R Kemler
Keyword(s):  
Cell Adhesion ◽  
Epithelial Cells ◽  
Adhesion Molecule ◽  
Cell Adhesion Molecule ◽  
Intestinal Epithelial Cells ◽  
Rabbit Antiserum ◽  
Preimplantation Embryos ◽  
Intestinal Epithelial ◽  
Electron Microscopic ◽  
A Cell

Uvomorulin is a cell-adhesion molecule implicated in the compaction process of mouse preimplantation embryos and the aggregation of embryonal carcinoma cells. A rabbit antiserum against purified uvomorulin also reacts with epithelial cells of various adult tissues. In this study, we investigated the localization of uvomorulin on adult intestinal epithelial cells using electron microscopic analyses. Uvomorulin was shown to exhibit a highly restricted localization in the intermediate junctions of these cells. The results are discussed with respect to a possible adhesive function of uvomorulin on intestinal epithelial cells.

scholarly journals Salmonella Type III Effector AvrA Stabilizes Cell Tight Junctions to Inhibit Inflammation in Intestinal Epithelial Cells

PLoS ONE ◽  
2008 ◽  
Vol 3 (6) ◽  
pp. e2369 ◽  
Author(s):  
Anne P. Liao ◽  
Elaine O. Petrof ◽  
Sumalatha Kuppireddi ◽  
Yun Zhao ◽  
Yinglin Xia ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Tight Junctions ◽  
Intestinal Epithelial Cells ◽  
Intestinal Epithelial ◽  
Type Iii Effector ◽  
Type Iii

scholarly journals Intermediate Filaments Interact with Dormant Ezrin in Intestinal Epithelial Cells

2005 ◽  
Vol 16 (9) ◽  
pp. 4096-4107 ◽  
Author(s):  
Flavia A. Wald ◽  
Andrea S. Oriolo ◽  
M. Llanos Casanova ◽  
Pedro J.I. Salas
Keyword(s):  
Epithelial Cells ◽  
Intermediate Filaments ◽  
Brush Border ◽  
Antisense Rna ◽  
Apical Membrane ◽  
Intestinal Epithelial Cells ◽  
Intestinal Epithelial ◽  
Undifferentiated Cells ◽  
Responsive Element

Ezrin connects the apical F-actin scaffold to membrane proteins in the apical brush border of intestinal epithelial cells. Yet, the mechanisms that recruit ezrin to the apical domain remain obscure. Using stable CACO-2 transfectants expressing keratin 8 (K8) antisense RNA under a tetracycline-responsive element, we showed that the actin-ezrin scaffold cannot assemble in the absence of intermediate filaments (IFs). Overexpression of ezrin partially rescued this phenotype. Overexpression of K8 in mice also disrupted the assembly of the brush border, but ezrin distributed away from the apical membrane in spots along supernumerary IFs. In cytochalasin D-treated cells ezrin localized to a subapical compartment and coimmunoprecipitated with IFs. Overexpression of ezrin in undifferentiated cells showed a Triton-insoluble ezrin compartment negative for phospho-T567 (dormant) ezrin visualized as spots along IFs. Pulse-chase analysis showed that Triton-insoluble, newly synthesized ezrin transiently coimmunoprecipitates with IFs during the first 30 min of the chase. Dormant, but not active (p-T567), ezrin bound in vitro to isolated denatured keratins in Far-Western analysis and to native IFs in pull-down assays. We conclude that a transient association to IFs is an early step in the polarized assembly of apical ezrin in intestinal epithelial cells.

scholarly journals JAM4, a Junctional Cell Adhesion Molecule Interacting with a Tight Junction Protein, MAGI-1

2003 ◽  
Vol 23 (12) ◽  
pp. 4267-4282 ◽  
Author(s):  
Susumu Hirabayashi ◽  
Makiko Tajima ◽  
Ikuko Yao ◽  
Wataru Nishimura ◽  
Hiroki Mori ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Epithelial Cells ◽  
Tight Junctions ◽  
Adhesion Molecule ◽  
Intestinal Epithelial Cells ◽  
Cell Junctions ◽  
Intestinal Epithelial ◽  
Cell Contacts ◽  
L Cells ◽  
Junction Protein

ABSTRACT MAGI-1 is a membrane-associated guanylate kinase protein at tight junctions in epithelial cells. It interacts with various molecules and functions as a scaffold protein at cell junctions. We report here a novel MAGI-1-binding protein that we named junctional adhesion molecule 4 (JAM4). JAM4 belongs to an immunoglobulin protein family. JAM4 was colocalized with ZO-1 in kidney glomeruli and in intestinal epithelial cells. Biochemical in vitro studies revealed that JAM4 bound to MAGI-1 but not to ZO-1, whereas JAM1 did not bind to MAGI-1. JAM4 and MAGI-1 interacted with each other and formed clusters in COS-7 cells when coexpressed. JAM4 mediated calcium-independent homophilic adhesion and was accumulated at cell-cell contacts when expressed in L cells. MAGI-1, ZO-1, and occludin were recruited to JAM4-based cell contacts. JAM4 also reduced the permeability of CHO cell monolayers. MAGI-1 strengthened JAM4-mediated cell adhesion in L cells and sealing effects in CHO cells. These findings suggest that JAM4 together with MAGI-1 provides an adhesion machinery at tight junctions, which may regulate the permeability of kidney glomerulus and small intestinal epithelial cells.

scholarly journals The function of macromolecular complex of CFTR-NHERF2-LPA2 in inflammatory responses of intestinal epithelial cells

2017 ◽  
Author(s):  
Shanshan Kong ◽  
Weiqiang Zhang
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Line ◽  
Human Cell ◽  
Inflammatory Responses ◽  
Intestinal Epithelial Cells ◽  
Human Cell Line ◽  
Macromolecular Complex ◽  
Apical Surface ◽  
Intestinal Epithelial

AbstractCFTR is a cAMP-regulated chloride channel located in the apical surface of intestinal epithelial cells; where it forms a macromolecular complex with NHERF2 and LPA2. CFTR has been shown to play a role in the pathogenies of several types of secretory diarrheas. Inflammatory bowel disease (IBD) is a chronic condition of intestine characterized by severe inflammation and mucosal destruction, genetic analysis has shown that LPA contribute to IBD and patients of cystic fibrosis also display the phenotype of diarrhea. The purpose of this study is to investigate if this complex plays a role in the inflammatory responses of intestinal epithelium.We then explored the role of this complex in maintaining the integrity of tight junction and inflammatory responses in these cells. In vitro assays show that inhibiting CFTR or LPA2 in the intestinal epithelial cell could disrupt the epithelial cell junction, and reduce the TER of intestinal epithelial cells in both mouse and human cell line. EƯSA assay show that intriguing LPA2 through LPS or LPA can increase the secretion of IL-8, while inhibiting or SiRNA knockdown of LPA2 can decrease the secretion of IL-8 in mouse or human intestinal epithelial cells. The CFTR inhibitor can reduce the IL-8 secretion in both mouse and human cell line, the deletion of CFTR in mouse intestine does not affect the IL-8 level, but the knockdown of CFTR in human cell line reduced the IL-8 protein level. The deletion of CFTR in human also reduced the IL-8 mRNA level. This indicates the CFTR-LPA complex is necessary for the expression of IL-8.

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