‘Putting the squeeze’ on the tight junction: understanding cytoskeletal regulation

doi:10.1006/scdb.2000.0180

Seminars in Cell & Developmental Biology

Volume 11, Issue 4, August 2000, Pages 301-308

https://doi.org/10.1006/scdb.2000.0180 Get rights and content

Abstract

The apical perijunctional actomyosin ring of epithelia is structurally associated with the tight junction. The functional association between the tight junction and the perijunctional actomyosin ring was initially described in studies using pharmacological agents that disrupt microfilaments. More recently, this interaction has been studied in physiological, pathophysiological, and molecular models of tight junction regulation. These studies have demonstrated the central role of actomyosin contraction in tight junction regulation. With the identification of novel tight junction proteins and characterization of their protein:protein interactions comes the promise of detailed understanding of the molecular interactions that mediate tight junction regulation.

References (41)

JL Madara
Intestinal absorptive cell tight junctions are linked to cytoskeleton
Am J Physiol
(1987)
JL Madara et al.
Effects of cytochalasin D on occluding junctions of intestinal absorptive cells: further evidence that the cytoskeleton may influence paracellular permeability and junctional charge selectivity
J Cell Biol
(1986)
JL Madara et al.
Functional coupling of tight junctions and microfilaments in T84 monolayers
Am J Physiol
(1988)
BR Stevenson et al.
Concentration-dependent effects of cytochalasin D on tight junctions and actin filaments in MDCK epithelial cells
J Cell Sci
(1994)
CJ Bentzel et al.
Cytoplasmic regulation of tight-junction permeability: effect of plant cytokinins
Am J Physiol
(1980)
I Meza et al.
Occluding junctions and cytoskeletal components in a cultured transporting epithelium
J Cell Biol
(1980)
I Meza et al.
Occluding junctions in MDCK cells: modulation of transepithelial permeability by the cytoskeleton
J Cell Biochem
(1982)
JL Madara et al.
Alteration of intestinal tight junction structure and permeability by cytoskeletal contraction
Am J Physiol
(1987)
TY Ma et al.
Cytoskeletal regulation of Caco-2 intestinal monolayer paracellular permeability
J Cell Physiol
(1995)
JL Madara et al.
Structural basis for physiological regulation of paracellular pathways in intestinal epithelia
J Membr Biol
(1987)

JR Pappenheimer

Physiological regulation of transepithelial impedance in the intestinal mucosa of rats and hamsters

J Membr Biol

(1987)

K Atisook et al.

Effects of phlorizin and sodium on glucose-elicited alterations of cell junctions in intestinal epithelia

Am J Physiol

(1990)

JR Turner et al.

Physiological regulation of epithelial tight junctions is associated with myosin light-chain phosphorylation

Am J Physiol

(1997)

JR Turner et al.

Carboxy-terminal vesicular stomatitis virus G protein-tagged intestinal Na⁺-dependent glucose cotransporter (SGLT1): maintanence of surface expression and global transport function with selective perturbation of transport kinetics and polarized expression

J Biol Chem

(1996)

S Citi et al.

Regulation of non-muscle myosin structure and function

Bioessays

(1987)

JR Turner et al.

Protein kinase C-dependent regulation of transepithelial resistance: the roles of myosin light chain and myosin light chain kinase

Am J Physiol

(1999)

R Yuhan et al.

Enteropathogenic Escherichia coli-induced myosin light chain phosphorylation alters intestinal epithelial permeability

Gastroenterology

(1997)

ZM Goeckeler et al.

Myosin light chain kinase-regulated endothelial cell contraction: the relationship between isometric tension, actin polymerization, and myosin phosphorylation

J Cell Biol

(1995)

Y Yamaguchi et al.

Vasopressin and A23187 stimulate phosphorylation of myosin light chain-1 in isolated rat hepatocytes

Am J Physiol

(1991)

G Hecht et al.

Expression of the catalytic domain of myosin light chain kinase increases paracellular permeability

Am J Physiol

(1996)

Cited by (148)

Role of Bifidobacterium in Modulating the Intestinal Epithelial Tight Junction Barrier: Current Knowledge and Perspectives
2023, Current Developments in Nutrition
The intestinal tight junction (TJ) barrier is a crucial defense mechanism that prevents the passage of intestinal content into the intestinal wall, tissue, and systemic circulation. A compromised intestinal TJ barrier has been identified as a significant factor in inflammatory bowel disease (IBD), necrotizing enterocolitis, and other gut-related inflammatory conditions. Recent studies have revealed the importance of the probiotic bacterial strains of Bifidobacterium in protecting against intestinal inflammation and IBD pathogenesis via the regulation of intestinal TJ barrier function. Numerous species and strains of Bifidobacterium have been found to regulate TJ proteins and the signaling pathways responsible for maintaining intestinal barrier integrity and permeability. In this review, we provide a summary of recent studies that highlight the regulatory role of Bifidobacterium species and the strain effect on the intestinal TJ barrier. We also discuss the intracellular mechanisms involved in Bifidobacterium modulation of the intestinal barrier and the potential therapeutic efficacy of targeting the barrier function to regulate intestinal inflammation.
Effect of pH and lipopolysaccharide on tight junction regulators and inflammatory markers in intestinal cells as an experimental model of weaning transition in dairy calves
2023, JDS Communications
Acidic conditions combined with the presence of lipopolysaccharide (LPS) may increase the permeability of gastrointestinal epithelium. Feeding starch-rich starter to dairy calves is associated with ruminal acidosis and decreases the pH of other segments of the gastrointestinal tract, and that affects the tight junction regulator. The objective was to evaluate the effect of the combination of different pH (7.4 vs. 6.0) and LPS concentrations (0, 0.5, 10 ng/mL) in intestinal cells on tight junction regulators, inflammatory markers, and permeability. The human colon carcinoma Caco-2 cell line was used with the main treatment of pH and LPS in a 2 × 3 factorial arrangement. The pH was acidic (pH 6.0) or physiologic (pH 7.4), whereas LPS was 0, 0.5, or 10 ng/mL. After cells reached 70%–80% of confluence, the media were replaced with each respective treatment medium. Cells were treated for 3 h for mRNA abundance analysis, 3 and 6 h for protein abundance determination, and 3, 6, 12, and 24 h for permeability determination. Protein abundance of the myosin light-chain kinase (MYLK) and toll-like receptor 4 (TLR4) were measured by western blot. The mRNA abundance of IL-8, MYLK, peroxisome proliferator activated receptor gamma, and nuclear factor kappa B (Nfkb1) was determined by real-time, quantitative PCR. Paracellular permeability was determined with Lucifer yellow after 21 d of incubation. Cell culture was performed in biological triplicate; each biological replicate for real-time, quantitative PCR had 2 technical replicates, and for protein abundance and permeability assay had one technical replicate. The MIXED procedure of SAS (SAS Institute Inc.) was used with LPS, pH, and pH × LPS as fixed effects. Significance was declared at P ≤ 0.05 and tendencies when 0.05 < P ≤ 0.10. Increasing LPS doses did not affect the protein abundance of MYLK and TLR4, nor mRNA abundance of MYLK and PPRG. The LPS tended to increase mRNA abundance of IL-8 while pH × LPS interactively increased mRNA abundance of Nfkb1, where mRNA abundance of Nfkb1 was lower in cells exposed to pH 6.0 when combined with 0 and 10 ng/mL of LPS. Contrary to expectations, LPS did not affect the permeability of Caco-2 cells. The mRNA abundance of MYLK was greater at pH 6.0 versus pH 7.4. Also, protein abundance of TLR4 was lower at pH 6.0 than pH 7.4, and it decreased when exposure increased to 6 h. In addition, mRNA abundance of IL-8 was lower at pH 6.0 versus pH 7.4. Permeability was greater at pH 6.0 versus 7.4 after 6, 12, and 24 h of treatment. In summary, the effect of LPS and its interaction with pH showed less impact than expected on dependent variables measured, which might be attributed to the adopted clinically achievable LPS doses likely not being high enough to draw a strong response as observed in the literature. On the other hand, pH was far more relevant, modulating mRNA abundance of inflammatory markers, tight junction regulators, and permeability in in vitro colon cell models.
Analysis of absorption-enhancing mechanisms for combinatorial use of spermine with sodium taurocholate in Caco-2 cells
2022, European Journal of Pharmaceutics and Biopharmaceutics
Previously, we reported that the combined use of spermine (SPM) and sodium taurocholate (STC) (SPM–STC) significantly improves the oral absorption of rebamipide (BCS class IV) and pulmonary absorption of interferon-α without any harmful histopathological changes in the gastrointestinal tract and lungs, respectively. In the present study, we examined the effect of SPM–STC on the transport of fluorescein isothiocyanate-labeled dextrans (FDs) across Caco-2 cell monolayers and attempted to clarify the mechanisms underlying the transport enhancement caused by SPM–STC. SPM–STC were found to significantly enhance the transport of FDs, while the treatment with SPM–STC was not harmful, and the decrease in transepithelial electrical resistance was transient and reversible. The voltage-clamp study clearly indicated that the opening of the paracellular route could be mainly responsible for the enhanced transport of FD-4. As for the mechanisms, it was found that SPM–STC caused a significant increase in membrane fluidity, which would lead to the enhanced transport of small-molecule drugs such as rebamipide. Since SPM–STC increased intracellular Ca²⁺ via Ca²⁺ uptake through Ca²⁺ channels and Ca²⁺ release from the endoplasmic reticulum stimulated by the IP₃ pathway, the subsequent possible activation of the MLCK signaling pathway would have led to the contraction of the actin–myosin ring. The rearrangement of tight junction-constituting proteins induced through the MAPK pathway has also been suggested as a possible mechanism for opening tight junctions. Claudin-4, a key protein constituting the tight junction, merged with F-actin along with the plasma membrane, was significantly decreased, which would be at least partial structural evidence for the tight-junction opening.
Mechanisms of uptake and transport of particulate formulations in the small intestine
2022, Journal of Controlled Release
Citation Excerpt :
However, in certain cases, decrease in the integrity of tight junctions can lead to enlargement of the intercellular space, and in turn allow for the passage of particles. This enhanced permeation through tight junctional modulation has been observed to occur spontaneously when particles interact with epithelial cells as this may lead to decrease of the epithelial barrier integrity as shown in several in vivo studies [39,158–160]. The Caco-2 cells model was also used to explore the involvement of intercellular junctions in particle transport [158,159].
Micro- and nano-scale particulate formulations are widely investigated towards improving the oral bioavailability of both biologics and drugs with low solubility and/or low intestinal permeability. Particulate formulations harnessing physiological intestinal transport pathways have recently yielded remarkably high oral bioavailabilities, illustrating the need for better understanding the specific pathways underpinning particle small intestinal absorption and the relative role of intestinal cells. Mechanistic knowledge has been hampered by the well acknowledged limitations of current in vitro, in vivo and ex vivo models relevant to the human intestinal physiology and the lack of standardization in studies reporting absorption data. Here we review the relevant literature and critically discusses absorption pathways with a focus on the role of specific intestinal epithelial and immune cells. We conclude that while Microfold (M) cells are a valid target for oral vaccines, enterocytes play a greater role in the systemic bioavailability of orally administrated particulate formulations, particularly within the sub-micron size range. We also comment on less-reported mechanisms such as paracellular permeability of particles, persorption due to cell damage and uptake by migratory immune cells.
Tea polyphenol alleviate Aeromonas hydrophila - induced intestinal physical barrier damage in grass carp (Ctenopharyngodon idella)
2021, Aquaculture
This study aimed to investigate the effects of tea polyphenol (TP) supplementation on attenuating Aeromonas hydrophila - induced intestinal barrier function injury in fish. A total of 540 healthy grass carp (187.49 ± 0.66 g) were randomly divided into six groups with three replicates and fed TP-supplemented diets at 0, 40, 80, 120, 160 and 200 mg/kg for 60 days. After the growth trial, 24 fish per group were randomly selected for a challenge test with A. hydrophila for 6 days. The results indicated that an appropriate level of TP (120 mg/kg diet) repair damaged the intestinal morphology of fish induced by A. hydrophila. Additionally, appropriate level of TP ameliorated the intestinal apical junctional complex partly, which was partly related to the up-regulation of the expressions of tight junction and adherent junction proteins via the RhoA/ROCK pathway in the fish intestine. Based on the results of this research, dietary TP supplementation possesses a protective function of the intestinal physical barrier in fish against A. hydrophila, making TP a functional feed additive for aquaculture and providing a solution for the issue of hazards to human health caused by antibiotic residues in aquatic food.
Target specific tight junction modulators
2021, Advanced Drug Delivery Reviews
Intercellular tight junctions represent a formidable barrier against paracellular drug absorption at epithelia (e.g., nasal, intestinal) and the endothelium (e.g., blood–brain barrier). In order to enhance paracellular transport of drugs and increase their bioavailability and organ deposition, active excipients modulating tight junctions have been applied. First-generation of permeation enhancers (PEs) acted by unspecific interactions, while recently developed PEs address specific physiological mechanisms. Such target specific tight junction modulators (TJMs) have the advantage of a defined specific mechanism of action. To date, merely a few of these novel active excipients has entered into clinical trials, as their lack in safety and efficiency in vivo often impedes their commercialisation. A stronger focus on the development of such active excipients would result in an economic and therapeutic improvement of current and future drugs.

View all citing articles on Scopus

¹: Address for correspondence: 540 E. Canfield, 9248 Scott Hall, Detroit, MI 48201, USA. Email: [email protected]

View full text

Regular Article‘Putting the squeeze’ on the tight junction: understanding cytoskeletal regulation

Abstract

Intestinal absorptive cell tight junctions are linked to cytoskeleton

Am J Physiol

Effects of cytochalasin D on occluding junctions of intestinal absorptive cells: further evidence that the cytoskeleton may influence paracellular permeability and junctional charge selectivity

J Cell Biol

Functional coupling of tight junctions and microfilaments in T84 monolayers

Am J Physiol

Concentration-dependent effects of cytochalasin D on tight junctions and actin filaments in MDCK epithelial cells

J Cell Sci

Cytoplasmic regulation of tight-junction permeability: effect of plant cytokinins

Am J Physiol

Occluding junctions and cytoskeletal components in a cultured transporting epithelium

J Cell Biol

Occluding junctions in MDCK cells: modulation of transepithelial permeability by the cytoskeleton

J Cell Biochem

Alteration of intestinal tight junction structure and permeability by cytoskeletal contraction

Am J Physiol

Cytoskeletal regulation of Caco-2 intestinal monolayer paracellular permeability

J Cell Physiol

Structural basis for physiological regulation of paracellular pathways in intestinal epithelia

J Membr Biol

Physiological regulation of transepithelial impedance in the intestinal mucosa of rats and hamsters

J Membr Biol

Effects of phlorizin and sodium on glucose-elicited alterations of cell junctions in intestinal epithelia

Am J Physiol

Physiological regulation of epithelial tight junctions is associated with myosin light-chain phosphorylation

Am J Physiol

Carboxy-terminal vesicular stomatitis virus G protein-tagged intestinal Na+-dependent glucose cotransporter (SGLT1): maintanence of surface expression and global transport function with selective perturbation of transport kinetics and polarized expression

J Biol Chem

Regulation of non-muscle myosin structure and function

Bioessays

Protein kinase C-dependent regulation of transepithelial resistance: the roles of myosin light chain and myosin light chain kinase

Am J Physiol

Enteropathogenic Escherichia coli-induced myosin light chain phosphorylation alters intestinal epithelial permeability

Gastroenterology

Myosin light chain kinase-regulated endothelial cell contraction: the relationship between isometric tension, actin polymerization, and myosin phosphorylation

J Cell Biol

Vasopressin and A23187 stimulate phosphorylation of myosin light chain-1 in isolated rat hepatocytes

Am J Physiol

Expression of the catalytic domain of myosin light chain kinase increases paracellular permeability

Am J Physiol

Regular Article
‘Putting the squeeze’ on the tight junction: understanding cytoskeletal regulation

Carboxy-terminal vesicular stomatitis virus G protein-tagged intestinal Na⁺-dependent glucose cotransporter (SGLT1): maintanence of surface expression and global transport function with selective perturbation of transport kinetics and polarized expression