Rotavirus alters paracellular permeability and energy metabolism in Caco-2 cells

2000 ◽  
Vol 279 (4) ◽  
pp. G757-G766 ◽  
Author(s):  
Kathleen G. Dickman ◽  
Scott J. Hempson ◽  
Joseph Anderson ◽  
Scott Lippe ◽  
Liming Zhao ◽  
...  
Keyword(s):  
Tight Junction ◽  
Tight Junctions ◽  
Intestinal Cells ◽  
Metabolic Dysfunction ◽  
Rotavirus Infection ◽  
Severe Diarrhea ◽  
Consistent Feature ◽  
And Function ◽  
Junction Structure

Rotaviruses infect epithelial cells of the small intestine, but the pathophysiology of the resulting severe diarrhea is incompletely understood. Histological damage to intestinal epithelium is not a consistent feature, and in vitro studies showed that intestinal cells did not undergo rapid death and lysis during viral replication. We show that rotavirus infection of Caco-2 cells caused disruption of tight junctions and loss of transepithelial resistance (TER) in the absence of cell death. TER declined from 300 to 22 Ω · cm2between 8 and 24 h after infection and was accompanied by increased transepithelial permeability to macromolecules of 478 and 4,000 Da. Distribution of tight junction proteins claudin-1, occludin, and ZO-1 was significantly altered during infection. Claudin-1 redistribution was notably apparent at the onset of the decline in TER. Infection was associated with increased production of lactate, decreased mitochondrial oxygen consumption, and reduced cellular ATP (60% of control at 24 h after infection), conditions known to reduce the integrity of epithelial tight junctions. In conclusion, these data show that rotavirus infection of Caco-2 intestinal cells altered tight junction structure and function, which may be a response to metabolic dysfunction.

scholarly journals Direct Current Stimulation Disrupts Endothelial Glycocalyx and Tight Junctions of the Blood-Brain Barrier in vitro

2021 ◽  
Vol 9 ◽  
Author(s):  
Yifan Xia ◽  
Yunfei Li ◽  
Wasem Khalid ◽  
Marom Bikson ◽  
Bingmei M. Fu
Keyword(s):  
Endothelial Cells ◽  
Tight Junction ◽  
Tight Junctions ◽  
Blood Brain Barrier ◽  
Direct Current ◽  
Endothelial Glycocalyx ◽  
Microvascular Endothelial Cells ◽  
Bbb Permeability

Transcranial direct current stimulation (tDCS) is a non-invasive physical therapy to treat many psychiatric disorders and to enhance memory and cognition in healthy individuals. Our recent studies showed that tDCS with the proper dosage and duration can transiently enhance the permeability (P) of the blood-brain barrier (BBB) in rat brain to various sized solutes. Based on the in vivo permeability data, a transport model for the paracellular pathway of the BBB also predicted that tDCS can transiently disrupt the endothelial glycocalyx (EG) and the tight junction between endothelial cells. To confirm these predictions and to investigate the structural mechanisms by which tDCS modulates P of the BBB, we directly quantified the EG and tight junctions of in vitro BBB models after DCS treatment. Human cerebral microvascular endothelial cells (hCMECs) and mouse brain microvascular endothelial cells (bEnd3) were cultured on the Transwell filter with 3 μm pores to generate in vitro BBBs. After confluence, 0.1–1 mA/cm2 DCS was applied for 5 and 10 min. TEER and P to dextran-70k of the in vitro BBB were measured, HS (heparan sulfate) and hyaluronic acid (HA) of EG was immuno-stained and quantified, as well as the tight junction ZO-1. We found disrupted EG and ZO-1 when P to dextran-70k was increased and TEER was decreased by the DCS. To further investigate the cellular signaling mechanism of DCS on the BBB permeability, we pretreated the in vitro BBB with a nitric oxide synthase (NOS) inhibitor, L-NMMA. L-NMMA diminished the effect of DCS on the BBB permeability by protecting the EG and reinforcing tight junctions. These in vitro results conform to the in vivo observations and confirm the model prediction that DCS can disrupt the EG and tight junction of the BBB. Nevertheless, the in vivo effects of DCS are transient which backup its safety in the clinical application. In conclusion, our current study directly elucidates the structural and signaling mechanisms by which DCS modulates the BBB permeability.

scholarly journals Enteroaggregative Escherichia coli Disrupts Epithelial Cell Tight Junctions

2010 ◽  
Vol 78 (11) ◽  
pp. 4958-4964 ◽  
Author(s):  
Maura C. Strauman ◽  
Jill M. Harper ◽  
Susan M. Harrington ◽  
Erik Juncker Boll ◽  
James P. Nataro
Keyword(s):  
Escherichia Coli ◽  
Tight Junction ◽  
Tight Junctions ◽  
Cell Model ◽  
Barrier Dysfunction ◽  
T84 Cells ◽  
E Coli ◽  
Eaec Strain ◽  
Epithelial Barrier Dysfunction

ABSTRACT Enteroaggregative Escherichia coli (EAEC) is responsible for inflammatory diarrhea in diverse populations, but its mechanisms of pathogenesis have not been fully elucidated. We have used a previously characterized polarized intestinal T84 cell model to investigate the effects of infection with EAEC strain 042 on tight junction integrity. We find that infection with strain 042 induces a decrease in transepithelial electrical resistance (TER) compared to uninfected controls and to cells infected with commensal E. coli strain HS. When the infection was limited after 3 h by washing and application of gentamicin, we observed that the TER of EAEC-infected monolayers continued to decline, and they remained low even as long as 48 h after the infection. Cells infected with the afimbrial mutant strain 042aafA exhibited TER measurements similar to those seen in uninfected monolayers, implicating the aggregative adherence fimbriae II (AAF/II) as necessary for barrier dysfunction. Infection with wild-type strain 042 induced aberrant localization of the tight junction proteins claudin-1 and, to a lesser degree, occludin. EAEC-infected T84 cells exhibited irregular shapes, and some cells became elongated and/or enlarged; these effects were not observed after infection with commensal E. coli strain HS or 042aafA. The effects on tight junctions were also observed with AAF/I-producing strain JM221, and an afimbrial mutant was similarly deficient in inducing barrier dysfunction. Our results show that EAEC induces epithelial barrier dysfunction in vitro and implicates the AAF adhesins in this phenotype.

The tight junction: Structure and function

Micron ◽  
1993 ◽  
Vol 24 (3) ◽  
pp. 325-352 ◽  
Author(s):  
Michel Hirsch ◽  
Walter Noske
Keyword(s):  
Tight Junction ◽  
Structure And Function ◽  
And Function ◽  
Junction Structure

scholarly journals Do Tight Junction Structure and Function Play a Role in the Acid Resistance of Barrett’s Esophagus?

2007 ◽  
Vol 21 (6) ◽  
Author(s):  
Biljana Jovov ◽  
Christina M. Van Itallie ◽  
James M. Anderson ◽  
Roy C. Orlando
Keyword(s):  
Tight Junction ◽  
Barrett’S Esophagus ◽  
Barrett's Esophagus ◽  
Acid Resistance ◽  
Structure And Function ◽  
And Function ◽  
Junction Structure

Phosphorylation of occludin correlates with occludin localization and function at the tight junction

1997 ◽  
Vol 273 (6) ◽  
pp. C1859-C1867 ◽  
Author(s):  
Vivian Wong
Keyword(s):  
Molecular Weight ◽  
Tight Junction ◽  
Tight Junctions ◽  
Growth Medium ◽  
Mdck Cells ◽  
Low Calcium ◽  
Phosphatase Treatment ◽  
And Function ◽  
Triton X ◽  
Mdck I

Multiple forms of occludin were found in Madin-Darby canine kidney (MDCK) cells. In the absence of cell-to-cell contacts, achieved by incubating cells in low-calcium growth medium, a cluster of lower-molecular-weight (LMW) occludin bands (∼65,000–68,000) was present in both MDCK I and II cells. On formation of tight junctions, achieved by changing the low-calcium growth medium to normal-calcium growth medium, a cluster of higher-molecular-weight (HMW) bands (∼72,000–75,000 for MDCK I cells and ∼70,000–73,000 for MDCK II cells) was also expressed. The HMW occludin bands could be eliminated by phosphatase treatment. Therefore, the HMW forms of occludin appeared to be the hyperphosphorylated product of the LMW forms. These HMW forms were Triton X-100 insoluble, which correlated with their localization at the tight junctions. Furthermore, depletion of tight junction-localized occludin by an occludin extracellular domian peptide (20) correlated with a decrease in the HMW forms of occludin. In conclusion, phosphorylation of occludin may be a mechanism by which occludin localization and function are regulated.

scholarly journals The Unique-5 and -6 Motifs of ZO-1 Regulate Tight Junction Strand Localization and Scaffolding Properties

2007 ◽  
Vol 18 (3) ◽  
pp. 721-731 ◽  
Author(s):  
Alan S. Fanning ◽  
Brent P. Little ◽  
Christoph Rahner ◽  
Darkhan Utepbergenov ◽  
Zenta Walther ◽  
...  
Keyword(s):  
Tight Junction ◽  
Tight Junctions ◽  
Cultured Cells ◽  
Variable Region ◽  
Binding Studies ◽  
Vitro Binding ◽  
Complex Protein ◽  
Regulatory Functions ◽  
Maguk Proteins

The proper cellular location and sealing of tight junctions is assumed to depend on scaffolding properties of ZO-1, a member of the MAGUK protein family. ZO-1 contains a conserved SH3-GUK module that is separated by a variable region (unique-5), which in other MAGUKs has proven regulatory functions. To identify motifs in ZO-1 critical for its putative scaffolding functions, we focused on the SH3-GUK module including unique-5 (U5) and unique-6 (U6), a motif immediately C-terminal of the GUK domain. In vitro binding studies reveal U5 is sufficient for occludin binding; U6 reduces the affinity of this binding. In cultured cells, U5 is required for targeting ZO-1 to tight junctions and removal of U6 results in ectopically displaced junction strands containing the modified ZO-1, occludin, and claudin on the lateral cell membrane. These results provide evidence that ZO-1 can control the location of tight junction transmembrane proteins and reveals complex protein binding and targeting signals within its SH3-U5-GUK-U6 region. We review these findings in the context of regulated scaffolding functions of other MAGUK proteins.

Sign in / Sign up

Export Citation Format

Share Document