Substratum influence on collagen and fibronectin biosynthesis by arterial smooth muscle cells in vitro

Holderbaum, Daniel; Ehrhart, L A

doi:10.1002/jcp.1041260210

Cited by 35 publications

(12 citation statements)

References 33 publications

(32 reference statements)

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“…High doses yielded FN synthesis inhibition, while low doses elicited a modest increase. These results are in agreement with those obtained in arterial smooth muscle cells, where FN adhesion regulates FN synthesis in a negative manner [21,22]. Likewise, FN administration to animals with immune chronic nephritis decreased glomerular FN synthesis [23].…”

Section: Discussionsupporting

confidence: 91%

The 80-kD fibronectin fragment increases the production of fibronectin and tumour necrosis factor-alpha (TNF-α) in cultured mesangial cells

López-Armada

González

Gómez-Guerrero

et al. 1997

Clinical and Experimental Immunology

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SUMMARYThe presence of fibronectin (FN) fragments has been demonstrated in several inflammatory processes, and they have been implicated in the recruitment of mononuclear cells. However, the interaction of these FN fragments with resident cells has hardly been studied. We have hypothesized that the 80-kD FN fragment, which includes the RGD cell binding domain of FN, could contribute to the pathogenesis of glomerular damage through the interaction with mesangial cells (MC) via a 5 b 1 integrin. Since an increase in the glomerular deposit of matrix components, particularly FN, is frequently observed in progressive glomerulonephritis, we studied whether its synthesis is modulated by the 80-kD FN fragment and the native FN molecule. While the 80-kD FN fragment stimulated FN in a dose-dependent manner, both at the mRNA and protein level, the whole FN molecule exerted a dual effect. High doses produced FN inhibition, while low doses elicited a certain increase. This stimulation was abrogated by the presence of Sam-1, a MoAb against the a-subunit of the a 5 b 1 integrin. Since cytokines play a fundamental role in glomerular injury, we studied the production of TNF-a, one of the most powerful mediators of inflammation. TNF-a synthesis was induced by the 80-kD FN fragment, in a dosedependent manner, but not by native FN. When MC were incubated with the 29-and 31-kD FN fragments, which lack the RGD cell binding domain, TNF-a secretion was not detected. These results strongly suggest that in cultured MC, the 80-kD FN fragment induces the synthesis of matrix proteins such as FN, and cytokines such as TNF-a, via a 5 b 1 integrin. This mechanism could contribute to the perpetuation of renal injury.

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Section: Discussionsupporting

confidence: 91%

The 80-kD fibronectin fragment increases the production of fibronectin and tumour necrosis factor-alpha (TNF-α) in cultured mesangial cells

López-Armada

González

Gómez-Guerrero

et al. 1997

Clinical and Experimental Immunology

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“…Recently, preconfluent cultures of arterial smooth muscle cells on FN-coated substrates were shown to synthesize 50% less fibronectin compared with similar cultures on albumin-coated surfaces (17). This observation suggests that the cells are capable of recognizing specific extracellular matrix molecules and appropriately regulating their synthesis via a negative feedback mechanism.…”

Section: Discussionmentioning

confidence: 99%

The fibronectin cell attachment sequence Arg-Gly-Asp-Ser promotes focal contact formation during early fibroblast attachment and spreading.

Singer¹,

Kawka²,

Scott³

et al. 1987

The Journal of Cell Biology

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Abstract. Cultured fibroblasts form focal contacts (FCs) associated with actin microfilament bundles (MFBs) during attachment and spreading on serum-or fibronectin (FN)-coated substrates. To determine if the minimum cellular adhesion receptor recognition signal Arg-Gly-Asp-Ser (RGDS) is sufficient to promote FC and MFB formation, rat (NRK), hamster (Nil 8), and mouse (Balb/c 3T3) fibroblasts in serum-free media were plated on substrates derivatized with small synthetic peptides containing RGDS. These cultures were studied with interference reflection microscopy to detect FCs, Normarski optics to identify MFBs, and immunofluorescence microscopy to observe endogenous FN fiber formation. By 1 h, 72-78 % of the NRK and Nil 8 cells plated on RGDS-containing peptide had focal contacts without accompanying FN fibers, while these fibroblasts lacked FCs on control peptide. This early FC formation was followed by the appearance of coincident MFBs and colinear FN fbers forming fibronexuses at 4 h. NRK and Nil 8 cultures on substrates coated with native FN or 75,000-D FN-cell binding fragment showed similar kinetics of FC and MFB formation. In contrast, the Balb/c 3T3 mouse fibroblasts plated on Gly-Arg-Gly-Asp-Ser peptidederivatized substrates, or on coverslips coated with 75,000-D FN cell-binding fragment, were defective in FC formation. These results demonstrate that the apparent binding of substrate-linked RGDS sequences to cell surface adhesion receptors is sufficient to promote early focal contact formation followed by the appearance of fibronexuses in some, but not all, fibroblast lines. F IBRONECTIN (FN) ~ is a large glycoprotein found in plasma and the connective tissue extracellular matrix, which functions primarily in promoting cellular adhesion to solid substrates. It is composed of several protease-resistant functional domains that can bind independently to a number of important substrates, including fibrin, collagen, heparin, and various cell surface components (18,35). The FN cell-attachment domain has recently been shown to bind to a 140-kD FN receptor complex (1, 2, 33) that codistributes with microfilament bundles at cell-substrate adhesion sites (4, 5). Through the use of small synthetic peptides deduced from the primary structure of the FN cellattachment domain, the tetrapeptide Arg-Gly-Asp-Ser has been found to be the minimal FN sequence with cell attachment activity (29-31).Arg-Gly-Asp-Val, a sequence closely related to Arg-GlyAsp-Ser, is present in vitronectin, another protein with potent cell-adhesive properties that is localized with FN in the extracellular matrix (15,16,44). The vitronectin receptor has a different molecular mass than the FN receptor, and specifi-1. Abbreviations used in this paper: DIC, differential interference contrast; FC, focal contact(s); FN, fibronectin; HBSA, heat denatured BSA; IFM, immunofluorescence microscopy; IRM, interference reflection microscopy; SPDP, 3-(2-pyridol-dithio) propionic acid N-hydroxy-succinimide ester.cally binds vitronectin rather than FN. However, bo...

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“…4 " 6 Previous studies in our laboratory have demonstrated that preconfluent smooth muscle cells (SMCs) grown on fibronectin synthesize significantly less collagen and fibronectin than control SMCs, whereas noncollagenous protein synthesis was unaltered. 7 In contrast, both collagen and fibronectin syn-thesis by postconfluent cells was unaffected by the substratum. Quantification of collagen synthesis and procollagen mRNA levels in SMCs plated on an SMCderived matrix have not been previously reported.…”

Section: Abstract • Collagen • Smooth Muscle Cells • Basic Fibroblasmentioning

confidence: 97%

Basic fibroblast growth factor in the extracellular matrix suppresses collagen synthesis and type III procollagen mRNA levels in arterial smooth muscle cell cultures.

Majors

Ehrhart

1993

Arterioscler Thromb

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To determine the effects of an intact extracellular matrix on collagen synthesis, arterial smooth muscle cells (SMCs) were plated sparsely on a cell-free, SMC-derived matrix and examined the following day. Collagen synthesis during a 5-hour incubation by cells on the matrix was reduced to 67% of the control values obtained from cultures on plastic. Total protein synthesis was unaffected. Treatment of the matrix with heparitinase to remove basic fibroblast growth factor (bFGF) before seeding the SMCs abolished the inhibitory effect of the matrix on collagen synthesis. The inhibitory effect was also eliminated by treating the matrix with a neutralizing polyclonal antibody directed against bFGF. Collagen synthesis by SMC cultures grown in wells coated with purified bFGF was only 61% that of control cultures, whereas total protein synthesis remained unchanged. Slot-blot analysis revealed that the relative message level for orl(III) procollagen was reduced in cultures grown on the preexisting matrix or on plastic precoated with bFGF, whereas the al(I) procollagen message was unaffected. These results demonstrate the ability of the extracellular matrix to modulate the synthesis of collagen by arterial SMCs and indicate that bFGF in the matrix is responsible for these effects. (Arteriosclerosis and Thrombosis 1993; 13:680-686) KEY WORDS • collagen • smooth muscle cells • basic fibroblast growth factor • extracellular matrixC ells in vivo are in contact with an insoluble matrix composed of collagens, elastin, proteoglycans, noncollagenous glycoproteins, and a variety of adherent nonstructural components including growth factors. This extracellular matrix has the capacity to modulate the behavior and phenotype of cells both in vivo and in vitro. Several types of cultured cells have been shown to respond to the substratum onto which they are plated by altering their adhesion, growth, morphology, and protein synthesis patterns.Numerous studies have shown that preformed matrices, when used as a substratum for cultured cells, may have significant effects on cell proliferation.1 " 3 In addition, the substratum on which cells are plated can have specific effects on the synthesis of extracellular matrix components.4 " 6 Previous studies in our laboratory have demonstrated that preconfluent smooth muscle cells (SMCs) grown on fibronectin synthesize significantly less collagen and fibronectin than control SMCs, whereas noncollagenous protein synthesis was unaltered. 7 In contrast, both collagen and fibronectin syn-

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Substratum influence on collagen and fibronectin biosynthesis by arterial smooth muscle cells in vitro

Cited by 35 publications

References 33 publications

The 80-kD fibronectin fragment increases the production of fibronectin and tumour necrosis factor-alpha (TNF-α) in cultured mesangial cells

The 80-kD fibronectin fragment increases the production of fibronectin and tumour necrosis factor-alpha (TNF-α) in cultured mesangial cells

The fibronectin cell attachment sequence Arg-Gly-Asp-Ser promotes focal contact formation during early fibroblast attachment and spreading.

Basic fibroblast growth factor in the extracellular matrix suppresses collagen synthesis and type III procollagen mRNA levels in arterial smooth muscle cell cultures.

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