scholarly journals Cellular zinc is required for intestinal epithelial barrier maintenance via the regulation of claudin-3 and occludin expression

2016 ◽  
Vol 311 (1) ◽  
pp. G105-G116 ◽  
Author(s):  
Yuka Miyoshi ◽  
Soichi Tanabe ◽  
Takuya Suzuki
Keyword(s):  
Electrical Resistance ◽  
Molecular Mechanisms ◽  
Transepithelial Electrical Resistance ◽  
Surface Proteins ◽  
Pcr Analysis ◽  
Intestinal Epithelial ◽  
Calpain Activity ◽  
Intracellular Zinc ◽  
Cell Functions ◽  
Zinc Depletion

Intracellular zinc is required for a variety of cell functions, but its precise roles in the maintenance of the intestinal tight junction (TJ) barrier remain unclear. The present study investigated the essential roles of intracellular zinc in the preservation of intestinal TJ integrity and the underlying molecular mechanisms. Depletion of intracellular zinc in both intestinal Caco-2 cells and mouse colons through the application of a cell-permeable zinc chelator N, N, N′, N′-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) induced a disruption of the TJ barrier, as indicated by increased FITC-labeled dextran flux and decreased transepithelial electrical resistance. The TPEN-induced TJ disruption is associated with downregulation of two TJ proteins, occludin and claudin-3. Biotinylation of cell surface proteins revealed that the zinc depletion induced the proteolysis of occludin but not claudin-3. Occludin proteolysis was sensitive to the inhibition of calpain activity, and increased calpain activity was observed in the zinc-depleted cells. Although quantitative PCR analysis and promoter reporter assay have demonstrated that the zinc depletion-induced claudin-3 downregulation occurred at transcriptional levels, a site-directed mutation in the egr1 binding site in the claudin-3 promoter sequence induced loss of both the basal promoter activity and the TPEN-induced decreases. Reduced egr1 expression by a specific siRNA also inhibited claudin-3 expression and transepithelial electrical resistance maintenance in cells. This study shows that intracellular zinc has an essential role in the maintenance of the intestinal epithelial TJ barrier through regulation of occludin proteolysis and claudin-3 transcription.

scholarly journals Application of Measurements of Transepithelial Electrical Resistance of Intestinal Epithelial Cell Monolayers To Evaluate Probiotic Activity

2005 ◽  
Vol 71 (11) ◽  
pp. 7528-7530 ◽  
Author(s):  
Trine Danø Klingberg ◽  
Maja Herold Pedersen ◽  
Avrelija Cencic ◽  
Birgitte Bjørn Budde
Keyword(s):  
Electrical Resistance ◽  
Intestinal Epithelial Cell ◽  
Transepithelial Electrical Resistance ◽  
Intestinal Epithelial ◽  
Lactobacillus Salivarius ◽  
Cell Monolayers ◽  
Epithelial Cell Monolayers ◽  
Probiotic Lactobacilli ◽  
Dose Dependent ◽  
Probiotic Activity

ABSTRACT Among five potentially probiotic lactobacilli investigated, Lactobacillus plantarum MF1298 and Lactobacillus salivarius DC5 showed the highest increase in the transepithelial electrical resistance (TER) of polarized monolayers of Caco-2 cells, and this increase was shown to be dose dependent. Furthermore, preincubation with MF1298 attenuated a decrease in TER induced by Listeria monocytogenes.

scholarly journals Desmoglein2 Regulates Claudin2 Expression by Sequestering PI-3-Kinase in Intestinal Epithelial Cells

2021 ◽  
Vol 12 ◽  
Author(s):  
Natalie Burkard ◽  
Michael Meir ◽  
Felix Kannapin ◽  
Christoph Otto ◽  
Maximilian Petzke ◽  
...  
Keyword(s):  
Electrical Resistance ◽  
Intestinal Epithelial Cells ◽  
Direct Interaction ◽  
Transepithelial Electrical Resistance ◽  
Intestinal Epithelial ◽  
Intestinal Epithelial Barrier ◽  
Proximity Ligation ◽  
Inflammatory Conditions ◽  
Basal Expression ◽  
Caco2 Cells

Inflammation-induced reduction of intestinal desmosomal cadherin Desmoglein 2 (Dsg2) is linked to changes of tight junctions (TJ) leading to impaired intestinal epithelial barrier (IEB) function by undefined mechanisms. We characterized the interplay between loss of Dsg2 and upregulation of pore-forming TJ protein Claudin2. Intraperitoneal application of Dsg2-stablising Tandem peptide (TP) attenuated impaired IEB function, reduction of Dsg2 and increased Claudin2 in DSS-induced colitis in C57Bl/6 mice. TP blocked loss of Dsg2-mediated adhesion and upregulation of Claudin2 in Caco2 cells challenged with TNFα. In Dsg2-deficient Caco2 cells basal expression of Claudin2 was increased which was paralleled by reduced transepithelial electrical resistance and by augmented phosphorylation of AKTSer473 under basal conditions. Inhibition of phosphoinositid-3-kinase proved that PI-3-kinase/AKT-signaling is critical to upregulate Claudin2. In immunostaining PI-3-kinase dissociated from Dsg2 under inflammatory conditions. Immunoprecipitations and proximity ligation assays confirmed a direct interaction of Dsg2 and PI-3-kinase which was abrogated following TNFα application. In summary, Dsg2 regulates Claudin2 expression by sequestering PI-3-kinase to the cell borders in intestinal epithelium.

scholarly journals 124 Butyric acid and derivatives: In vitro effects on barrier integrity of porcine intestinal epithelial cells quantified by transepithelial electrical resistance

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 109-110
Author(s):  
Lauren L Kovanda ◽  
Monika Hejna ◽  
Yanhong Liu
Keyword(s):  
Butyric Acid ◽  
Sodium Butyrate ◽  
Barrier Function ◽  
Electrical Resistance ◽  
Intestinal Epithelial Cells ◽  
Intestinal Barrier ◽  
Transepithelial Electrical Resistance ◽  
Intestinal Barrier Function ◽  
Intestinal Epithelial

Abstract Intestinal barrier function in vitro is quantified by the transepithelial electrical resistance (TEER) across epithelial cell monolayers due to polarization and expression of tight junction proteins. The objective of the current study was to measure the TEER of porcine intestinal epithelial cells (IPEC-J2) treated with butyric acid, sodium butyrate, monobutyrin and tributyrin. MTT assays were performed for each compound to determine cell viability and appropriate treatment doses. Butyric acid and tributyrin treatments were: 0, 0.5, 1, 2, and 4 mM. Monobutyrin and sodium butyrate doses were: 0, 1, 2, 4, 8 mM. Cells were seeded at 5 × 105 cells/mL into 12-well plates on Corning transwell inserts. Cells were cultured for 4 to 5 days (until all wells had TEER close to 1000 Ωcm2) and were then treated with organic acids. Each compound was tested in a randomized complete block design with 10 replicates. TEER was measured in Ωcm2 using a Millicell ERS-2 voltohmmeter at 0 h (before treatment) and at 24, 48, and 72 h post-treatment. All data were analyzed by PROC MIXED of SAS. Butyric acid linearly enhanced (P < 0.001) TEER of IPEC-J2 dose-dependently, with the highest TEER observed at doses of 1 and 2 mM at each time point. Sodium butyrate did not impact TEER at h 24, but linearly increased (P < 0.05) TEER at h 48 and 72. Treatment of monobutyrin linearly improved (P < 0.001) TEER at h 24, 48, and 72, with the highest TEER observed at the dose of 8 mM. Tributyrin had the tendency to improve (P < 0.10) TEER at h 48 and 72 when the highest dose was applied. Results of the current in vitro study indicate that butyric acid and its derivatives may improve intestinal barrier function of pigs, which requires verification in vivo.

scholarly journals Enterococcus faeciumHDRsEf1 Protects the Intestinal Epithelium and Attenuates ETEC-Induced IL-8 Secretion in Enterocytes

2016 ◽  
Vol 2016 ◽  
pp. 1-10 ◽  
Author(s):  
Zhongyuan Tian ◽  
Xiaofang Liu ◽  
Ran Dai ◽  
Yuncai Xiao ◽  
Xiliang Wang ◽  
...  
Keyword(s):  
Electrical Resistance ◽  
Cell Layer ◽  
Intestinal Epithelial Cells ◽  
Intestinal Barrier ◽  
Transepithelial Electrical Resistance ◽  
Therapeutic Interventions ◽  
Intestinal Epithelial ◽  
Cell Free Supernatant ◽  
Positive Effects ◽  
Intestinal Pathogen

The probioticEnterococcus faeciumHDRsEf1 (Ef1) has been shown to have positive effects on piglet diarrhoea, but the mechanism has not yet been elucidated. In this study, using the IPEC-J2 cell line to mimic intestinal epithelial cells and enterotoxigenicEscherichia coli(ETEC) K88ac as a representative intestinal pathogen, the mechanism underlying Ef1 protection against an enteropathogen was investigated. The results demonstrated that Ef1 was effective in displacing K88ac from the IPEC-J2 cell layer. Moreover, Ef1 and its cell-free supernatant (S-Ef1) modulate IL-8 released by IPEC-J2 cells. Ef1 and its cell-free supernatant showed the potential to protect enterocytes from an acute inflammatory response. In addition, Ef1 and its cell-free supernatant increased the transepithelial electrical resistance (TEER) of the enterocyte monolayer, thus strengthening the intestinal barrier against ETEC. These results may contribute to the development of therapeutic interventions using Ef1 in intestinal disorders of piglets.

scholarly journals Protective Effect of QiOne® 2 Pro on Cultured Intestinal Epithelial Cells after Mobile Phone Radiation

2021 ◽  
Vol 9 (3) ◽  
Author(s):  
Peter C Dartsch
Keyword(s):  
Oxidative Stress ◽  
Coherent State ◽  
Mobile Phone ◽  
Electrical Resistance ◽  
Intestinal Epithelial Cells ◽  
Transepithelial Electrical Resistance ◽  
Epithelial Barrier ◽  
Water Molecules ◽  
Intestinal Epithelial ◽  
Mobile Phone Radiation

QiOne® 2 Pro is a specific device which creates a static field that stimulates water molecules to undergo a transition into the coherent state. Since our body consists of about 70 to 85% of water (depending on age), this coherent state of the water molecules might increase the cellular resistance against exogenous reliabilities such as electromagnetic fields. In this study, the protective effect of QiOne® 2 Pro against mobile phone radiation was examined by using the cultured intestinal epithelial cells. Unprotected cells and untreated control cells served as point of reference. The cell regeneration process as well as the integrity of the intestinal epithelial barrier was investigated by measuring the transepithelial electrical resistance. Mobile phone radiation caused a reduced cell regenerative activity by approximately 60%, whereas the values were about 15% for QiOne® 2 Pro protected cells and untreated controls, respectively. Moreover, mobile phone radiation caused a rupture on the epithelial barrier in unprotected cells by cell death caused due to the oxidative stress with a complete loss of morphological integrity on the barrier. In contrast, untreated controls and QiOne® 2 Pro protected cells did not show any morphological change on the cell layers with an epithelial barrier of a 10-fold higher transepithelial electrical resistance than the unprotected cells. Overall the results clearly demonstrate the sensitivity of intestinal barrier against oxidative stress generated by mobile phone radiation. In addition, the results also show that the QiOne® 2 Pro device is able to reduce unwanted cellular effects of mobile phone radiation.

scholarly journals Revising Endosomal Trafficking under Insulin Receptor Activation

2021 ◽  
Vol 22 (13) ◽  
pp. 6978
Author(s):  
Maria J. Iraburu ◽  
Tommy Garner ◽  
Cristina Montiel-Duarte
Keyword(s):  
Plasma Membrane ◽  
Tyrosine Kinase ◽  
Insulin Receptor ◽  
Molecular Mechanisms ◽  
Receptor Activation ◽  
Small Gtpases ◽  
Early Endosome ◽  
Tyrosine Kinase Receptors ◽  
Endosomal Trafficking ◽  
Cell Functions

The endocytosis of ligand-bound receptors and their eventual recycling to the plasma membrane (PM) are processes that have an influence on signalling activity and therefore on many cell functions, including migration and proliferation. Like other tyrosine kinase receptors (TKR), the insulin receptor (INSR) has been shown to be endocytosed by clathrin-dependent and -independent mechanisms. Once at the early endosome (EE), the sorting of the receptor, either to the late endosome (LE) for degradation or back to the PM through slow or fast recycling pathways, will determine the intensity and duration of insulin effects. Both the endocytic and the endosomic pathways are regulated by many proteins, the Arf and Rab families of small GTPases being some of the most relevant. Here, we argue for a specific role for the slow recycling route, whilst we review the main molecular mechanisms involved in INSR endocytosis, sorting and recycling, as well as their possible role in cell functions.

scholarly journals Common pathways and functional profiles reveal underlying patterns in Breast, Kidney and Lung cancers

2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Sergio Romera-Giner ◽  
Zoraida Andreu Martínez ◽  
Francisco García-García ◽  
Marta R. Hidalgo
Keyword(s):  
Molecular Mechanisms ◽  
Machine Learning Techniques ◽  
Alternative Activation ◽  
Survival Times ◽  
Major Health Problem ◽  
Lung Cancers ◽  
Cancer Data ◽  
Cell Functions ◽  
Cancer Pathogenesis ◽  
Tissue Of Origin

Abstract Background Cancer is a major health problem which presents a high heterogeneity. In this work we explore omics data from Breast, Kidney and Lung cancers at different levels as signalling pathways, functions and miRNAs, as part of the CAMDA 2019 Hi-Res Cancer Data Integration Challenge. Our goal is to find common functional patterns which give rise to the generic microenvironment in these cancers and contribute to a better understanding of cancer pathogenesis and a possible clinical translation down further studies. Results After a tumor versus normal tissue comparison of the signaling pathways and cell functions, we found 828 subpathways, 912 Gene Ontology terms and 91 Uniprot keywords commonly significant to the three studied tumors. Such features interestingly show the power to classify tumor samples into subgroups with different survival times, and predict tumor state and tissue of origin through machine learning techniques. We also found cancer-specific alternative activation subpathways, such as the ones activating STAT5A in ErbB signaling pathway. miRNAs evaluation show the role of miRNAs, such as mir-184 and mir-206, as regulators of many cancer pathways and their value in prognoses. Conclusions The study of the common functional and pathway activities of different cancers is an interesting approach to understand molecular mechanisms of the tumoral process regardless of their tissue of origin. The existence of platforms as the CAMDA challenges provide the opportunity to share knowledge and improve future scientific research and clinical practice.

scholarly journals Endurance Training Regulates Expression of Some Angiogenesis-Related Genes in Cardiac Tissue of Experimentally Induced Diabetic Rats

Biomolecules ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 498
Author(s):  
Mojdeh Khajehlandi ◽  
Lotfali Bolboli ◽  
Marefat Siahkuhian ◽  
Mohammad Rami ◽  
Mohammadreza Tabandeh ◽  
...  
Keyword(s):  
Gene Expression ◽  
Endurance Training ◽  
Molecular Mechanisms ◽  
Cardiac Tissue ◽  
Critical Role ◽  
Diabetic Rats ◽  
Moderate Intensity ◽  
Pcr Analysis ◽  
Cardiac Tissues ◽  
The Impact

Exercise can ameliorate cardiovascular dysfunctions in the diabetes condition, but its precise molecular mechanisms have not been entirely understood. The aim of the present study was to determine the impact of endurance training on expression of angiogenesis-related genes in cardiac tissue of diabetic rats. Thirty adults male Wistar rats were randomly divided into three groups (N = 10) including diabetic training (DT), sedentary diabetes (SD), and sedentary healthy (SH), in which diabetes was induced by a single dose of streptozotocin (50 mg/kg). Endurance training (ET) with moderate-intensity was performed on a motorized treadmill for six weeks. Training duration and treadmill speed were increased during five weeks, but they were kept constant at the final week, and slope was zero at all stages. Real-time polymerase chain reaction (RT-PCR) analysis was used to measure the expression of myocyte enhancer factor-2C (MEF2C), histone deacetylase-4 (HDAC4) and Calmodulin-dependent protein kinase II (CaMKII) in cardiac tissues of the rats. Our results demonstrated that six weeks of ET increased gene expression of MEF2C significantly (p < 0.05), and caused a significant reduction in HDAC4 and CaMKII gene expression in the DT rats compared to the SD rats (p < 0.05). We concluded that moderate-intensity ET could play a critical role in ameliorating cardiovascular dysfunction in a diabetes condition by regulating the expression of some angiogenesis-related genes in cardiac tissues.

scholarly journals High throughput transepithelial electrical resistance (TEER) measurements on perfused membrane-free epithelia

Lab on a Chip ◽  
2021 ◽  
Author(s):  
A. Nicolas ◽  
F. Schavemaker ◽  
K. Kosim ◽  
D. Kurek ◽  
M. Haarmans ◽  
...  
Keyword(s):  
High Throughput ◽  
Electrical Resistance ◽  
Drug Exposure ◽  
Transepithelial Electrical Resistance ◽  
Ease Of Use ◽  
Inflammatory Processes

We present an instrument for simultaneously measuring TEER in up to 80 perfused epithelial tubules on an OrganoPlate. The sensitivity, speed and ease of use enables screening of tubules during formation, drug exposure and inflammatory processes.

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