Cell-type-specific fibronectin subunits generated by alternative splicing.

Paul, Jeremy I.; Schwarzbauer, Jean E.; Tamkun, John W.; Hynes, Richard O.

doi:10.1016/s0021-9258(18)67233-3

Cited by 134 publications

(39 citation statements)

References 53 publications

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“…We have previously characterized a model of proliferative glomerulonephritis, induced by HV, by a distinct temporal course involving mesangial cell migration, proliferation, and extracellular matrix synthesis (Barnes 1989; Barnes and Abboud 1993; Barnes et al 1994a,1995a, b). Our previous studies suggest that migrating mesangial cells do not require their own synthesis of Fn-EIIIA but may rely on exogenous sources of Fn isoforms derived from platelets and macrophages (Barnes et al 1994a,1995b), and agree with our in vitro studies (Barnes and Hevey 1991) showing a potent migratory mesangial cell response to platelet Fn, which is abundant in EIIIA (Paul et al 1986; Peters et al 1995). A switch to an α-SMA phenotype appeared to be related to mesangial cell synthesis of Fn-EIIIA and coincided with expression of α-SMA, proliferation, and matrix synthesis, suggesting that autocrine synthesis of Fn-EIIIA by mesangial cells has specific functions during the course of glomerular remodeling.…”

Section: Discussionsupporting

confidence: 88%

“…Synthesis of riboprobe, tissue preparation, in situ hybridization and autoradiography were identical to methods used previously (Barnes et al 1994a,1995a, b). Briefly, cDNA probes containing a 160- BP alternatively spliced region EIIIA [generously provided by Dr. Richard O. Hynes, Massachusetts Institute of Technology (Paul et al 1986)] and a 130- BP fragment derived from the 3'-untranslated rat α-SMA mRNA that is specific for α-SMA transcript (Kocher and Gabbiani 1987) (generously provided by Dr. Gabriel Gabbiani, University of Geneva) were used for generation of 35 S-labeled riboprobes to detect cellular localization of mRNA in sections of renal tissue. All experiments were performed simultaneously with the sense riboprobe as a negative control.…”

Section: Methodsmentioning

confidence: 99%

“…The functions of the EIIIA domain are not known. However, its close proximity to the RGDS cell binding domain suggests that this isoform has specific functional roles (Schwarzbauer et al 1985; Paul et al 1986; Hynes 1990; Schwarzbauer 1991; ffrench-Constant 1995). The Fn-EIIIA variant is abundantly expressed during embryogenesis (ffrench-Constant and Hynes 1988,1989; Peters and Hynes 1996) and at the margins of healing wounds (ffrench-Constant et al 1989; Brown et al 1993), whereas this domain is spliced out of plasma Fn (derived from hepatocytes) and many tissue-specific cells in adult tissues (Peters et al 1996), suggesting that it has important functions in remodeling (Paul et al 1986; Schwarzbauer 1991; ffrench-Constant 1995).…”

mentioning

confidence: 99%

“…However, its close proximity to the RGDS cell binding domain suggests that this isoform has specific functional roles (Schwarzbauer et al 1985; Paul et al 1986; Hynes 1990; Schwarzbauer 1991; ffrench-Constant 1995). The Fn-EIIIA variant is abundantly expressed during embryogenesis (ffrench-Constant and Hynes 1988,1989; Peters and Hynes 1996) and at the margins of healing wounds (ffrench-Constant et al 1989; Brown et al 1993), whereas this domain is spliced out of plasma Fn (derived from hepatocytes) and many tissue-specific cells in adult tissues (Peters et al 1996), suggesting that it has important functions in remodeling (Paul et al 1986; Schwarzbauer 1991; ffrench-Constant 1995). Functions for Fn-EIIIA have not been determined, but a close association with cells undergoing high rates of migration, proliferation, and differentiation suggests a role in cell activation (Schwarzbauer et al 1985; ffrench-Constant and Hynes 1988,1989; ffrench-Constant et al 1989; Hynes 1990; Schwarzbauer 1991; Brown et al 1993; Barnes et al 1994a,1995b; ffrench-Constant 1995; Peters and Hynes 1996).…”

mentioning

confidence: 99%

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Expression of Embryonic Fibronectin Isoform EIIIA Parallels α-Smooth Muscle Actin in Maturing and Diseased Kidney

Barnes

Musa

Mitchell

et al. 1999

J Histochem Cytochem.

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In this study we examined if an association exists between expression of an alternatively spliced "embryonic" fibronectin isoform EIIIA (Fn-EIIIA) and alpha-smooth muscle actin (alpha-SMA) in the maturing and adult rat kidney and in two unrelated models of glomerular disease, passive accelerated anti-glomerular basement membrane (GBM) nephritis and Habu venom (HV)-induced proliferative glomerulonephritis, using immunohistochemistry and in situ hybridization. Fn-EIIIA and alpha-SMA proteins were abundantly expressed in mesangium and in periglomerular and peritubular interstitium of 20-day embryonic and 7-day (D-7) postnatal kidneys in regions of tubule and glomerular development. Staining was markedly reduced in these structures in maturing juvenile (D-14) kidney and was largely lost in adult kidney. Expression of Fn-EIIIA and alpha-SMA was reinitiated in the mesangium and the periglomerular and peritubular interstitium in both models and was also observed in glomerular crescents in anti-GBM nephritis. Increased expression of Fn-EIIIA mRNA by in situ hybridization corresponded to the localization of protein staining. Dual labeling experiments verified co-localization of Fn-EIIIA and alpha-SMA, showing a strong correlation of staining between location and staining intensity during kidney development, maturation, and disease. Expression of EIIIA mRNA corresponded to protein expression in developing and diseased kidneys and was lost in adult kidney. These studies show a recapitulation of the co-expression of Fn-EIIIA and alpha-SMA in anti-GBM disease and suggest a functional link for these two proteins.

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Section: Discussionsupporting

confidence: 88%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Expression of Embryonic Fibronectin Isoform EIIIA Parallels α-Smooth Muscle Actin in Maturing and Diseased Kidney

Barnes

Musa

Mitchell

et al. 1999

J Histochem Cytochem.

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“…Fn also promotes cell migration and proliferation (Simonson et al 1989;Hynes 1990;Barnes and Hevey 1991) and is believed to be involved in cell matrix assembly (McDonald 1988;Hynes 1990;Morla et al 1994). Platelets and macrophages also secrete TSP-1 (Jaffe et al 1985;Frazier 1987) and Fn-EIIIA (Plow et al 1979;Alitalo et al 1980;Paul et al 1986) in amounts that could regulate these same cell behaviors in the local microenvironment and play a role in remodeling in wound healing and disease (Raugi and Lovett 1987;Reed et al 1993;DiPietro et al 1996).…”

mentioning

confidence: 99%

Differential Expression of Thrombospondin and Cellular Fibronectin During Remodeling in Proliferative Glomerulonephritis

Barnes

Mitchell

Kanalas

et al. 1999

J Histochem Cytochem.

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SUMMARY Thrombospondin-1 (TSP-1) and an alternatively spliced fibronectin (Fn)-EIIIA isoform are adhesive proteins associated with embryogenesis and tissue remodeling. We compared, by immunohistochemistry and in situ hybridization, the course of TSP-1 and Fn-EIIIA expression in a model of glomerulonephritis induced by Habu snake venom (HV) and characterized by mesangial cell migration, proliferation, and extracellular matrix (ECM) synthesis. At 24 hr after HV, TSP-1 and Fn-EIIIA proteins localized in the central aspects of lesions associated with platelets and macrophages and at the margins of lesions coinciding with mesangial cell migration (determined by Thy-1 staining). Mesangial cells at this time expressed TSP-1 but not Fn-EIIIA mRNA. TSP-1 protein and mRNA peaked in lesions at 48 hr and were associated with cell proliferation (determined by PCNA, ␣-smooth muscle actin phenotype, and expression of ␤-PDGF receptor mRNA). TSP-1 expression declined at 72 hr when expression of ECM synthesis peaked, as determined by increased expression of collagen Type IV, laminin, and TGF-␤ 1 protein and mRNA. Mesangial cell expression of Fn-EIIIA was first observed at 48 hr and was most abundant at 72 hr after HV. Therefore, plateletand macrophage-derived Fn-EIIIA and TSP-1 in early lesions are associated with mesangial cell migration. Mesangial cell upregulation of TSP-1 is associated with migration and proliferation but not maximal ECM accumulation, whereas mesangial cell expression of Fn-EIIIA is associated with proliferation and ECM accumulation. These results suggest distinctive temporal and spatial roles for TSP-1 and Fn-EIIIA in remodeling during glomerular disease. Thrombospondin-1 (TSP-1) and cellular fibronectin, particularly the alternatively spliced isoform containing an extradomain EIIIA (Fn-EIIIA), are highly expressed during embryogenesis (ffrench-Constant and

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Gene expression and synthesis of fibronectin isoforms in rat hepatic stellate cells. comparison with liver parenchymal cells and skin fibroblasts

Xu¹,

Niki²,

Virtanen

et al. 1997

J. Pathol.

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Fibronectins are multifunctional glycoproteins that are important components of the extracellular matrix in normal and fibrotic liver. Multiple fibronectin isoforms are generated from a single gene by alternative splicing of the primary transcript at the domains EIIIA, EIIIB, and V. The aim of this study was to investigate the fibronectin isoforms expressed by activated hepatic stellate cells, the most important connective tissue‐producing cells in injured liver. Hepatocytes and skin fibroblasts were also studied for comparison. Activation of hepatic stellate cells in vivo was induced by injecting CCl4 twice weekly for 3 weeks. Activation in vitro was achieved by culturing cells on plastic. The level of activation was evaluated by α‐smooth muscle actin immunocytochemistry. Steady‐state levels of fibronectin isoform messenger RNA were examined by Northern hybridization analysis using specific cDNA probes for the EIIIA, EIIIB, and V domains. The de novo synthesis of fibronectin isoforms was examined by metabolic labelling and immunoprecipitation using domain‐specific monoclonal antibodies. Fibronectin transcripts were not detectable in freshly isolated hepatic stellate cells from normal liver. Cultured hepatic stellate cells, as well as skin fibroblasts, expressed EIIIA+, EIIIB+ and V95+ transcripts. They were detectable as early as day 3 and increased with time in culture. At 3 days in culture, more than 90 per cent of stellate cells were α‐smooth muscle actin‐positive. In vivo activated hepatic stellate cells expressed EIIIA+ and V95+ transcripts; EIIIB+ fibronectin mRNA was absent. Less than 20 per cent of in vivo activated stellate cells expressed α‐smooth muscle actin. Freshly isolated parenchymal cells from normal liver as well as from CCl4‐treated liver expressed V95+ transcripts, but were negative for EIIIA or EIIIB fibronectin mRNA. Immunoprecipitation results were in accordance with Northern hybridization analysis. Hepatic stellate cells in culture synthesized and secreted fibronectin molecules that contained EIIIA, EIIIB, and V fragments. Our results indicate that hepatic stellate cells synthesize and secrete fibronectin isoforms that are distinct from those of parenchymal cells. © 1997 by John Wiley & Sons, Ltd.

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Cell-type-specific fibronectin subunits generated by alternative splicing.

Cited by 134 publications

References 53 publications

Expression of Embryonic Fibronectin Isoform EIIIA Parallels α-Smooth Muscle Actin in Maturing and Diseased Kidney

Expression of Embryonic Fibronectin Isoform EIIIA Parallels α-Smooth Muscle Actin in Maturing and Diseased Kidney

Differential Expression of Thrombospondin and Cellular Fibronectin During Remodeling in Proliferative Glomerulonephritis

Gene expression and synthesis of fibronectin isoforms in rat hepatic stellate cells. comparison with liver parenchymal cells and skin fibroblasts

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