Detection of glycosaminoglycans on the surface of human umbilical vein endothelial cells using gold-conjugated poly-l-lysine with silver enhancement

Klein, Nigel; Shennan, Graham I.; Heyderman, Robert S.; Levin, Michael

doi:10.1007/bf00159120

Cited by 21 publications

(13 citation statements)

References 37 publications

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“…To study whether HS loss also causes intestinal protein leakage in vivo, we established 2 independent methods to assess enteric protein loss in mice, one determining fecal loss of 51 Cr-labeled albumin, the other measuring fecal levels of α1-antitrypsin (AAT) (10). The 2 methods gave comparable results, varying less than 10%.…”

Section: Sdc1 Knockdown Causes Protein Leakage In Vitromentioning

confidence: 99%

“…Heparin (lane 2) and 2/3-DS-H (lane 9) were most effective in alleviating cytokine-induced protein leakage (arrows). Lane 1, HS; lane 2, high-molecular-weight heparin (unfractionated); lane 3, low-molecular-weight heparin; lane 4, sized heparin, dp 2; lane 5, sized heparin, dp 8; lane 6, sized heparin, dp 14; lane 7, sized heparin, dp 20; 51 Cr labeling. Mice were injected daily with low (100 U/kg, 0.7 mg/kg) (B and E) or high doses (500 U/kg, 3.5 mg/kg) of heparin (C and F), 2/3-DS-H (C, D, F, and G), or PBS as control.…”

Section: Figurementioning

confidence: 99%

“…Sulfated GAG distribution was localized on formalin-fixed tissues as previously reported (22,51). Briefly, cationic colloidal gold (polylysine gold; Biocell International) was suspended 1:100 in PBS at pH 1.2 and applied to the tissue sections for 1 h at room temperature.…”

Section: Figurementioning

confidence: 99%

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Heparan sulfate and syndecan-1 are essential in maintaining murine and human intestinal epithelial barrier function

Bode

Salvestrini²,

Park³

et al. 2008

J. Clin. Invest.

133

125

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Patients with protein-losing enteropathy (PLE) fail to maintain intestinal epithelial barrier function and develop an excessive and potentially fatal efflux of plasma proteins. PLE occurs in ostensibly unrelated diseases, but emerging commonalities in clinical observations recently led us to identify key players in PLE pathogenesis. These include elevated IFN-γ, TNF-α, venous hypertension, and the specific loss of heparan sulfate proteoglycans from the basolateral surface of intestinal epithelial cells during PLE episodes. Here we show that heparan sulfate and syndecan-1, the predominant intestinal epithelial heparan sulfate proteoglycan, are essential in maintaining intestinal epithelial barrier function. Heparan sulfate-or syndecan-1-deficient mice and mice with intestinal-specific loss of heparan sulfate had increased basal protein leakage and were far more susceptible to protein loss induced by combinations of IFN-γ, TNF-α, and increased venous pressure. Similarly, knockdown of syndecan-1 in human epithelial cells resulted in increased basal and cytokine-induced protein leakage. Clinical application of heparin has been known to alleviate PLE in some patients but its unknown mechanism and severe side effects due to its anticoagulant activity limit its usefulness. We demonstrate here that non-anticoagulant 2,3-de-O-sulfated heparin could prevent intestinal protein leakage in syndecan-deficient mice, suggesting that this may be a safe and effective therapy for PLE patients.

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Section: Sdc1 Knockdown Causes Protein Leakage In Vitromentioning

confidence: 99%

Section: Figurementioning

confidence: 99%

See 1 more Smart Citation

Heparan sulfate and syndecan-1 are essential in maintaining murine and human intestinal epithelial barrier function

Bode

Salvestrini²,

Park³

et al. 2008

J. Clin. Invest.

133

125

View full text Add to dashboard Cite

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“…Anionic sites (mainly representing sulphated glycosaminoglycans, the major constituents of the ECM) were visualized with a gold-conjugated poly- L -lysine probe [11, 12]. Deparaffinized sections were rinsed in PBS (pH 2.0) for 10 min, then incubated on a drop of cationic (poly- L -lysine) colloidal gold (5 nm) (diluted 1:100 in PBS at pH 2.0; Biocell Research Laboratories, Cardiff, UK) for 30 min.…”

Section: Methodsmentioning

confidence: 99%

Mechanisms and Kinetics of Bowman’s Epithelial-Myofibroblast Transdifferentiation in the Formation of Glomerular Crescents

Fujigaki¹,

Sun²,

Fujimoto³

et al. 2002

Nephron

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Background: We investigated the mechanisms and kinetics of Bowman’s epithelial-myofibroblast transdifferentiation in the formation of glomerular crescents. Methods: Crescentic glomerulonephritis was induced by i.v. injection of rabbit anti-rat glomerular basement membrane antiserum in WKY rats. Results: Cellular crescents (83.5% of glomeruli) were first observed at day 7 after disease induction. Immunostaining of alpha-smooth muscle actin (alpha-SMA), as a marker for the myofibroblast phenotype, was found in some periglomerular regions as early as day 3, when it was also seen in parietal epithelial cells (PEC) of Bowman’s capsule at day 5 and in crescent formation at day 7. Proliferation marker Ki67-positive PEC was found at day 3, and double Ki67- and alpha-SMA-positive PEC could be seen at day 5. The migratory figure of PEC with the expression of alpha-SMA was found by immunoelectron microscopy. At day 7, some crescent cells were stained positive for PEC marker, protein gene product 9.5, in association with alpha-SMA or Ki67. Expression of transforming growth factor (TGF)-β receptor types I and II, as well as platelet-derived growth factor (PDGF) receptor β and PDGF-B increased in PEC as early as day 3. At day 5 marked deposition of cellular and common fibronectin, but not other extracellular matrix components examined was found in Bowman’s spaces where ED 1-positive macrophages infiltrated. Conclusions: PEC may be stimulated to proliferate and/or transdifferentiate into myofibroblast phenotype possibly by action of TGF-β and PDGF and/or binding of fibronectin to PEC, then migrate and/or proliferate, participating in glomerular crescents.

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“…Anionic sites (mainly representing sulfated glycosaminoglycans, the major constituents of the ECM) were visualized with a goldconjugated poly-l-lysine probe [15]. Deparaffinized sections were rinsed in PBS (pH 2.0) for 10 min, then incubated on a drop of cationic (poly-l-lysine) colloidal gold (5 nm) (diluted 1:100 in PBS at pH 2.0; Biocell Rescarch Laboratories, Cardiff, UK) for 4 h at 37C.…”

Section: Cationic Gold Stainingmentioning

confidence: 99%

Important role for fibronectin-EIIIA during renal tubular repair and cellular recovery in uranyl acetate-induced acute renal failure of rats

et al. 2003

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The present study was designed to identify the source and kinetics of an alternatively spliced "embryonic" cellular fibronectin EIIIA (cFn-EIIIA) in relation to regenerating renal tubules in uranyl acetate (UA)-induced acute renal failure (ARF) in rats. Damage of the proximal tubules was found as early as day 2 after induction of ARF, peaked at day 5, and was almost substituted by epithelial relining by day 7. Immunohistochemistry showed de novo deposition of cFn-EIIIA in peritubular regions as early as day 2, then on the tubular basement membrane (TBM) after day 4. beta1 Integrin, the receptor for Fn, was mainly found at the basal side of tubules in the normal control and increased in the interstitium after induction of ARF, but the staining pattern gradually returned to the control after day 7. Immunoelectron microscopy revealed that cFn-EIIIA was produced initially by the peritubular endothelium and later by fibroblastic cells and was deposited to the TBM, on which regenerating tubules proliferated, probably with cFn-EIIIA production. beta1 Integrin was expressed in cFn-EIIIA-producing cells, especially in regenerating tubular cells, suggesting that cFn-EIIIA signal transduction affects regenerating tubules. Transforming growth factor (TGF)-beta1 was found in some damaged proximal tubules and interstitial cells after induction of ARF and later in the regenerating tubules. CFn-EIIIA and beta1 integrin mRNA levels were upregulated as early as day 2. TGF-beta1 mRNA level significantly increased after day 3, suggesting a modulatory role for TGF-beta1 on cFn-EIIIA production, but not by day 2. Our data suggest that cFn-EIIIA production by the endothelium during the very early response to tubular injury and by fibroblastic cells and regenerating tubules may play an important role in the cellular recovery of UA-induced ARF in rats.

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Detection of glycosaminoglycans on the surface of human umbilical vein endothelial cells using gold-conjugated poly-l-lysine with silver enhancement

Cited by 21 publications

References 37 publications

Heparan sulfate and syndecan-1 are essential in maintaining murine and human intestinal epithelial barrier function

Heparan sulfate and syndecan-1 are essential in maintaining murine and human intestinal epithelial barrier function

Mechanisms and Kinetics of Bowman’s Epithelial-Myofibroblast Transdifferentiation in the Formation of Glomerular Crescents

Important role for fibronectin-EIIIA during renal tubular repair and cellular recovery in uranyl acetate-induced acute renal failure of rats

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