Expression of J1/tenascin in the crypt-villus unit of adult mouse small intestine: implications for its role in epithelial cell shedding

Development ◽  
1990 ◽  
Vol 109 (2) ◽  
pp. 313-321 ◽  
Author(s):  
R. Probstmeier ◽  
R. Martini ◽  
M. Schachner
Keyword(s):  
Extracellular Matrix ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Basal Lamina ◽  
Adult Mouse ◽  
Epithelial Cell Line ◽  
Intestinal Epithelial ◽  
Collagen Types ◽  
Numerous Microvillus ◽  
Cell Shedding

The localization of the extracellular matrix recognition molecule J1/tenascin was investigated in the crypt-villus unit of the adult mouse ileum by immunoelectron microscopic techniques. In the villus region, J1/tenascin was detected strongly in the extracellular matrix (ECM) between fibroblasts of the lamina propria. It was generally absent in the ECM at the interface between subepithelial fibroblasts and intestinal epithelium, except for some restricted areas along the epithelial basal lamina of villi, but not of crypts. These restricted areas corresponded approximately to the basal part of one epithelial cell. In J1/tenascin-positive areas, epithelial cells contacted the basal lamina with numerous microvillus-like processes, whereas in J1/tenascin-negative areas the basal surface membranes of epithelial cells contacted their basal lamina in a smooth and continuous apposition. In order to characterize the functional role of J1/tenascin in the interaction between epithelial cells and ECM, the intestinal epithelial cell line HT-29 was tested for its ability to adhere to different ECM components. Cells adhered to substratum-immobilized fibronectin, laminin and collagen types I to IV, but not to J1/tenascin. When laminin or collagen types I to IV were mixed with J1/tenascin, cell adhesion was as effective as without J1/tenascin. However, adhesion was completely abolished when cells were offered a mixture of fibronectin and J1/tenascin as substratum. The ability of J1/tenascin to reduce the adhesion of intestinal epithelial cells to their fibronectin-containing basal lamina suggests that J1/tenascin may be involved in the process of physiological cell shedding from the villus.

Studying Permeability in a Commonly Used Epithelial Cell Line: T84 Intestinal Epithelial Cells

2011 ◽  
pp. 115-137 ◽  
Author(s):  
Rino P. Donato ◽  
Adaweyah El-Merhibi ◽  
Batjargal Gundsambuu ◽  
Kai Yan Mak ◽  
Emma R. Formosa ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cell Line ◽  
Intestinal Epithelial Cells ◽  
Epithelial Cell Line ◽  
Intestinal Epithelial

47 Burn-induced Ileal Extracellular Matrix Upregulation Associated with Epithelial to Mesenchymal Transition

2020 ◽  
Vol 41 (Supplement_1) ◽  
pp. S31-S31
Author(s):  
Claire B Cummins ◽  
Xiaofu Wang ◽  
Yanping Gu ◽  
Jusong Song ◽  
Ravi S Radhakrishnan
Keyword(s):  
Extracellular Matrix ◽  
Epithelial Cells ◽  
Western Blot ◽  
Burn Injury ◽  
Total Body Surface Area ◽  
Epithelial Cell Line ◽  
Intestinal Epithelial ◽  
Mesenchymal Transition ◽  
Intestinal Fibrosis ◽  
Junction Protein

Abstract Introduction Severe burns have long been associated with systemic inflammation and intestinal dysfunction. Recent evidence suggests that intestinal fibrosis may be responsible for intestinal dysfunction after burns. In response to inflammatory stimuli, intestinal epithelial cells may undergo epithelial-mesenchymal transition (EMT). EMT is a process by which epithelial cells acquire a mesenchymal-like phenotype, thereby compromising epithelial barrier function. EMT has been implicated in the pathogenesis of intestinal fibrosis. In the present study, we examined the cellular mechanism of burn-induced intestinal dysfunction. Methods Male BALA/c mice (8–12 weeks) received 30% total body surface area full-thickness scald burns or sham procedure. Ileal tissue was collected 5 days after burn for immunofluorescence (IF) and Western blot. Rat intestinal epithelial cell line IEC-6 was treated with cytokines and EMT marker proteins were analyzed by Western blot. Results IF data demonstrated that burn significantly increased extracellular matrix (ECM) protein laminin in ileal tissues, suggesting that burn injury induces ileal fibrogenesis. In IEC-6 cells treated with Tumor necrosis factor (TNF) α, IL-1β dose-dependently upregulated α-smooth muscle actin (α-SMA), a well known marker for myofibroblasts. ECM proteins fibronectin and laminin were found to be significantly increased after TNFα treatment. Tight junction protein E-cadherin was decreased after TNFα treatment. Futhermore, TNF receptor signaling antagonist R-7050 and SM7368 blocked TNFα-induced α-SMA upregulation. Conclusions Burn-induced mouse ileum ECM upregulation may be through TNFα-mediated EMT. Applicability of Research to Practice TNFα receptor antagonism could represent a potential pathway for drug development for treatment of burn-induced intestinal dysfunction.

scholarly journals Mechanism of cholera toxin action on a polarized human intestinal epithelial cell line: role of vesicular traffic

1992 ◽  
Vol 117 (6) ◽  
pp. 1197-1209 ◽  
Author(s):  
WI Lencer ◽  
C Delp ◽  
MR Neutra ◽  
JL Madara
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cholera Toxin ◽  
Cell Line ◽  
Intestinal Epithelial Cell ◽  
Time Course ◽  
Epithelial Cell Line ◽  
Intestinal Epithelial ◽  
Vesicular Traffic ◽  
Intestinal Epithelial Cell Line

The massive secretion of salt and water in cholera-induced diarrhea involves binding of cholera toxin (CT) to ganglioside GM1 in the apical membrane of intestinal epithelial cells, translocation of the enzymatically active A1-peptide across the membrane, and subsequent activation of adenylate cyclase located on the cytoplasmic surface of the basolateral membrane. Studies on nonpolarized cells show that CT is internalized by receptor-mediated endocytosis, and that the A1-subunit may remain membrane associated. To test the hypothesis that toxin action in polarized cells may involve intracellular movement of toxin-containing membranes, monolayers of the polarized intestinal epithelial cell line T84 were mounted in modified Ussing chambers and the response to CT was examined. Apical CT at 37 degrees C elicited a short circuit current (Isc: 48 +/- 2.1 microA/cm2; half-maximal effective dose, ED50 integral of 0.5 nM) after a lag of 33 +/- 2 min which bidirectional 22Na+ and 36Cl- flux studies showed to be due to electrogenic Cl- secretion. The time course of the CT-induced Isc response paralleled the time course of cAMP generation. The dose response to basolateral toxin at 37 degrees C was identical to that of apical CT but lag times (24 +/- 2 min) and initial rates were significantly less. At 20 degrees C, the Isc response to apical CT was more strongly inhibited (30-50%) than the response to basolateral CT, even though translocation occurred in both cases as evidenced by the formation of A1-peptide. A functional rhodamine-labeled CT-analogue applied apically or basolaterally at 20 degrees C was visualized only within endocytic vesicles close to apical or basolateral membranes, whereas movement into deeper apical structures was detected at 37 degrees C. At 15 degrees C, in contrast, reduction to the A1-peptide was completely inhibited and both apical and basolateral CT failed to stimulate Isc although Isc responses to 1 nM vasoactive intestinal peptide, 10 microM forskolin, and 3 mM 8Br-cAMP were intact. Re-warming above 32 degrees C restored CT-induced Isc. Preincubating monolayers for 30 min at 37 degrees C before cooling to 15 degrees C overcame the temperature block of basolateral CT but the response to apical toxin remained completely inhibited. These results identify a temperature-sensitive step essential to apical toxin action on polarized epithelial cells. We suggest that this event involves vesicular transport of toxin-containing membranes beyond the apical endosomal compartment.

Fibronectin fragments induce MMP activity in mouse mammary epithelial cells: evidence for a role in mammary tissue remodeling

2000 ◽  
Vol 113 (5) ◽  
pp. 795-806 ◽  
Author(s):  
P. Schedin ◽  
R. Strange ◽  
T. Mitrenga ◽  
P. Wolfe ◽  
M. Kaeck
Keyword(s):  
Extracellular Matrix ◽  
Mammary Gland ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Protease Activity ◽  
Mammary Epithelial Cells ◽  
Cell Loss ◽  
Mammary Epithelial ◽  
Epithelial Cell Line ◽  
Fibronectin Fragments

Mammary gland form and function are regulated by interactions between epithelium and extracellular matrix. Major glycoprotein components of extracellular matrix have been identified that give survival, proliferation and differentiation signals to mammary epithelial cells. We provide evidence that proteolytic fragments of the extracellular matrix glycoprotein, fibronectin, suppress growth and can promote apoptosis of mouse mammary epithelial cells. During mammary gland involution, total fibronectin and fibronectin fragment levels are increased. The peak levels of fibronectin protein and fragments are observed 4–6 days post-weaning, coincident with the peak in epithelial cell death. Using a model for hormone withdrawal-induced death of mammary epithelium, elevated levels of fibronectin proteolytic fragments were associated with apoptosis in TM-6 cells, a tumorigenic mouse mammary epithelial cell line. Treatment of TM-6 cells with exogenous fibronectin fragments (FN120) reduced cell number, and induced apoptosis and matrix degrading protease activity. Inhibition of matrix protease activity rescued TM-6 cell viability, indicating that FN120-induced cell loss is mediated through matrix protease activity. In a three-dimensional model for mammary gland development, FN120 reduced alveolar-like and promoted ductal-like development by a matrix protease-dependent mechanism. These data suggest that during post-lactational involution, fibronectin fragments may contribute to epithelial cell loss and dissolution of mammary alveoli by inducing matrix degrading proteinases.

ADAM-15 inhibits wound healing in human intestinal epithelial cell monolayers

2005 ◽  
Vol 288 (2) ◽  
pp. G346-G353 ◽  
Author(s):  
Laetitia Charrier ◽  
Yutao Yan ◽  
Adel Driss ◽  
Christian L. Laboisse ◽  
Shanthi V. Sitaraman ◽  
...  
Keyword(s):  
Wound Healing ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Intestinal Epithelial Cell ◽  
Intestinal Epithelial Cells ◽  
Epithelial Cell Line ◽  
Intestinal Epithelial ◽  
Coding Region ◽  
Human Intestinal Epithelial Cell ◽  
New Variant

The disintegrin metalloproteases (or ADAMs) are membrane-anchored glycoproteins that have been implicated in cell-cell or cell-matrix interactions and in proteolysis of molecules on the cell surface. The expression and/or the pathophysiological implications of ADAMs are not known in intestinal epithelial cells. Therefore, our aim was to investigate the expression and the role of ADAMs in intestinal epithelial cells. Expression of ADAMs was assessed by RT-PCR, Western blot analysis, and immunufluorescence experiments. Wound-healing experiments were performed by using the electric cell substrate impedence sensing technology. Our results showed that ADAMs-10, -12, and -15 mRNA are expressed in the colonic human cell lines Caco2-BBE and HT29-Cl.19A. An ADAM-15 complementary DNA cloned from Caco2-BBE poly(A)+ RNA, and encompassing the entire coding region, was found to be shorter and to present a different region encoding the cytoplasmic tail compared with ADAM-15 sequence deposited in the database. In Caco2-BBE cells and colonic epithelial cells, ADAM-15 protein was found in the apical, basolateral, and intracellular compartments. We also showed that the overexpression of ADAM-15 reduced cell migration in a wound-healing assay in Caco2-BBE monolayers. Our data show that 1) ADAM-15 is expressed in human intestinal epithelia, 2) a new variant of ADAM-15 is expressed in a human intestinal epithelial cell line, and 3) ADAM-15 is involved in intestinal epithelial cells wound-healing processes. Together, these results suggest that ADAM-15 may have important pathophysiological roles in intestinal cells.

scholarly journals Bifidobacterium breve reduces apoptotic epithelial cell shedding in an exopolysaccharide and MyD88-dependent manner

Open Biology ◽  
2017 ◽  
Vol 7 (1) ◽  
pp. 160155 ◽  
Author(s):  
K. R. Hughes ◽  
L. C. Harnisch ◽  
C. Alcon-Giner ◽  
S. Mitra ◽  
C. J. Wright ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Well Being ◽  
Dependent Manner ◽  
Intestinal Epithelial ◽  
Epithelial Barrier Function ◽  
Bifidobacterium Breve ◽  
Inflammatory Conditions ◽  
Inflammatory Bowel ◽  
Cell Shedding

Certain members of the microbiota genus Bifidobacterium are known to positively influence host well-being. Importantly, reduced bifidobacterial levels are associated with inflammatory bowel disease (IBD) patients, who also have impaired epithelial barrier function, including elevated rates of apoptotic extrusion of small intestinal epithelial cells (IECs) from villi—a process termed ‘cell shedding’. Using a mouse model of pathological cell shedding, we show that mice receiving Bifidobacterium breve UCC2003 exhibit significantly reduced rates of small IEC shedding. Bifidobacterial-induced protection appears to be mediated by a specific bifidobacterial surface exopolysaccharide and interactions with host MyD88 resulting in downregulation of intrinsic and extrinsic apoptotic responses to protect epithelial cells under highly inflammatory conditions. Our results reveal an important and previously undescribed role for B. breve , in positively modulating epithelial cell shedding outcomes via bacterial- and host-dependent factors, supporting the notion that manipulation of the microbiota affects intestinal disease outcomes.

scholarly journals Shiga Toxin 2e-Producing Escherichia coli Isolates from Humans and Pigs Differ in Their Virulence Profiles and Interactions with Intestinal Epithelial Cells

2005 ◽  
Vol 71 (12) ◽  
pp. 8855-8863 ◽  
Author(s):  
Anne-Katharina Sonntag ◽  
Martina Bielaszewska ◽  
Alexander Mellmann ◽  
Nadine Dierksen ◽  
Peter Schierack ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Shiga Toxin ◽  
Intestinal Epithelial Cell ◽  
Intestinal Epithelial Cells ◽  
Epithelial Cell Line ◽  
Uptake System ◽  
Intestinal Epithelial ◽  
E Coli

ABSTRACT Thirteen Escherichia coli strains harboring stx 2e were isolated from 11,056 human stools. This frequency corresponded to the presence of the stx 2e allele in 1.7% of all Shiga toxin-producing E. coli (STEC) strains. The strains harboring stx 2e were associated with mild diarrhea (n = 9) or asymptomatic infections (n= 4). Because STEC isolates possessing stx 2e are porcine pathogens, we compared the human STEC isolates with stx 2e-harboring E. coli isolated from piglets with edema disease and postweaning diarrhea. All pig isolates possessed the gene encoding the F18 adhesin, and the majority possessed adhesin involved in diffuse adherence; these adhesins were absent from all the human STEC isolates. In contrast, the high-pathogenicity island encoding an iron uptake system was found only in human isolates. Host-specific patterns of interaction with intestinal epithelial cells were observed. All human isolates adhered to human intestinal epithelial cell lines T84 and HCT-8 but not to pig intestinal epithelial cell line IPEC-J2. In contrast, the pig isolates completely lysed human epithelial cells but not IPEC-J2 cells, to which most of them adhered. Our data demonstrate that E. coli isolates producing Shiga toxin 2e have imported specific virulence and fitness determinants which allow them to adapt to the specific hosts in which they cause various forms of disease.

scholarly journals Interleukin (IL)-22 from IL-20 Subfamily of Cytokines Induces Colonic Epithelial Cell Proliferation Predominantly through ERK1/2 Pathway

2019 ◽  
Vol 20 (14) ◽  
pp. 3468 ◽  
Author(s):  
Md. Moniruzzaman ◽  
Ran Wang ◽  
Varinder Jeet ◽  
Michael A. McGuckin ◽  
Sumaira Z. Hasnain
Keyword(s):  
Cell Proliferation ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Imaging System ◽  
Intestinal Epithelial Cells ◽  
Human Colon ◽  
Epithelial Cell Line ◽  
Intestinal Epithelial ◽  
Human Colon Cancer ◽  
Transcriptional Changes

The interleukin (IL)-20 subfamily of cytokines consists of IL-19, IL-20, IL-22, IL-24, and IL-26, and the expression of IL-20, IL-22, and IL-24 is reported to be higher in the colon of patients with ulcerative colitis. Although the receptors for these cytokines are highly expressed in the colon epithelium, their effects on epithelial renewal are not clearly understood. This study evaluated the effects of IL-20, IL-22, and IL-24 in epithelial renewal using the LS174T human colon cancer epithelial cell line. LS174T cells were treated with IL-20, IL-22, and IL-24 (25, 50, and 100 ng/mL) and a live-cell imaging system was used to evaluate the effects on cell proliferation. Following treatment, the signaling pathways contributing to cell proliferation were investigated through Western blotting in LS174T cells and downstream transcriptional changes through qRT-PCR in LS174T cells, and RNA-Seq in primary murine intestinal epithelial cells. Our results demonstrated that only IL-22 promoted LS174T cell proliferation, mediated via extracellular-signal-regulated kinase (ERK)1/2-mediated downstream regulation of p90RSK, c-Jun, and transcriptional changes of TRIM15 and STOM. IL-22 also promoted expression of ERK1/2-independent genes such as DDR2, LCN2, and LRG1, which are known to be involved in cell proliferation and migration. This study suggests that IL-22 induces cell proliferation in highly proliferative cells such as intestinal epithelial cells.

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