Immunogold localization of procollagen III, fibronectin and heparan sulfate proteoglycan on ultrathin frozen sections of the normal rat liver

Geerts, Albert; Geuze, Hans J.; Slot, Jan W.; Voß, B.; Schuppan, Detlef; Schellinck, Paul; Wisse, Eduard

doi:10.1007/bf00482963

Cited by 66 publications

(25 citation statements)

References 27 publications

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“…Several studies have contributed to the characterization of the ECM, which is defined in vitro as the growth substratum remaining attached to culture dishes after removal of cells by Triton X-100 (3)(4)(5), EDTA (5), or cytochalasin B (6). Major components of the hepatic ECM include collagen type III, collagen IV, fibronectin, and laminin (2,7,8) as well as abundant basement membrane-type heparin sulfate proteoglycans (HSPG) (7,9,10). In the liver, the ECM is a discontinuous structure and is therefore unlikely to have an efficient filtration function as described for more organized matrices of other tissues (11).…”

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confidence: 99%

The HepG2 Extracellular Matrix Contains Separate Heparinase- and Lipid-releasable Pools of ApoE

Burgess

Gould

Marcel

1998

Journal of Biological Chemistry

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We have examined the association of apoE with the extracellular matrix (ECM) of HepG2 cells. Comparison of ECM prepared by previously published methods demonstrated that cytochalasin B-prepared material yielded the highest endogenous apoE, representing 23.6% of that in cell monolayers. ECM prepared with EDTA or Triton X-100 exhibited decreased levels of apoE, 3 and 6%, respectively. ECM bound very low density lipoprotein poorly (5-6% of the monolayer capacity); however, these incubations dramatically increased the apoE content of the ECM. Heparinase or suramin decreased apoE of the ECM by 19.6 and 37.3%, respectively, suggesting association with heparin sulfate proteoglycans. EDTA or EGTA also displaced 35% of the apoE, suggesting a Ca 2؉ -dependent association. Incubation with phosphatidylcholine vesicles (PCV) displaced 30% of the apoE, suggesting that lipid content affects association of apoE with the ECM. Data derived from sequential incubations with combinations of suramin, EGTA, and PCV were consistent with the presence of two distinct pools of apoE on the HepG2 ECM, one releasable with suramin and EGTA and the other releasable with lipids. Exogenously applied lipid-free apoE readily bound to the ECM; however, increasing the lipid content decreased its association. Lipid-free apoE could be equally displaced from the ECM with PCV or suramin. When lipid-free apoE adsorbed to microtiter wells was incubated with a triglyceride emulsion or palmitoyloleyl phosphatidylcholine micelles, the immunoreactivity of 3H1 (but not other antibodies), a monoclonal antibody against an epitope in the C-terminal domain of apoE, increased about 4-fold. In a similar manner, incubation of ECM with lipid dramatically increased the immunoreactivity of 3H1, indicating that apoE of the ECM exists in a lipid-poor form. Scatchard analysis demonstrated that the increased immunoreactivity was due to an increase in the number of antibody binding sites. In conclusion, the ECM contains two pools of lipid-poor apoE. One pool associates with the ECM through heparin sulfate proteoglycans-and Ca 2؉ -dependent interactions. A second pool of apoE dissociates from the ECM upon lipidation. The lipid-sensitive pool of apoE may participate in secretion or efflux of lipids or in the capture of lipoproteins by providing the apoE needed for receptor-mediated uptake.

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confidence: 99%

The HepG2 Extracellular Matrix Contains Separate Heparinase- and Lipid-releasable Pools of ApoE

Burgess

Gould

Marcel

1998

Journal of Biological Chemistry

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“…From these observations, it has been concluded that: (1) the fine reticular fibers are mainly composed of type III and V collagens, and (2) the collagen fibers and coarse reticular fibers in the periphery of liver lobules are composed of type I, III and V collagens. Simultaneous localization of type I collagen and type III collagen in the same cross-striated collagen fibril has also been revealed in the space of Disse of normal rat (Geerts et al, 1986(Geerts et al, , 1990. In either case, each ECM component localizes three-dimensionally in a restricted region in the normal liver lobule.…”

Section: Ecm In the Perisinusoidal Spacementioning

confidence: 71%

“…Normal perisinusoidal space contains type I, III, IV, V, and VI collagens, fibronectin, laminin, and proteoglycans (Martinez-Hernandez, 1984;Geerts et al, 1986Geerts et al, , 1990Geerts et al, , 1994Rojkind and Greenwel, 1994;Martinez-Hernandez andAmenta, 1993a, 1995). A matrix gradient in the space of Disse has been revealed by detailed and extensive ultrastructural and immunochemical studies Sigal et al, 1992).…”

Section: Ecm In the Perisinusoidal Spacementioning

confidence: 99%

“…Fibronectin is present mainly in amorphous material in close contact with the microvilli of the parenchymal cells (Geerts et al, 1986;Martinez-Hernandez andAmenta, 1993b, 1995). Heparan sulfate proteoglycan is present in diffuse material associated with the surface of the parenchymal and endothelial cells (Geerts et al, 1986).…”

Section: Ecm In the Perisinusoidal Spacementioning

confidence: 99%

“…Heparan sulfate proteoglycan is present in diffuse material associated with the surface of the parenchymal and endothelial cells (Geerts et al, 1986).…”

Section: Ecm In the Perisinusoidal Spacementioning

confidence: 99%

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Adhesion between Cells and Extracellular Matrix with Special Reference to Hepatic Stellate Cell Adhesion to Three-dimensional Collagen Fibers.

Imai

Sato

Senoo

2000

Cell Struct. Funct.

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ABSTRACT. Hepatic stellate cells are located in the perisinusoidal space (space of Disse), and extend their dendritic, thin membranous processes and fine fibrillar processes into this space. The stellate cells coexist with a three-dimensional extracellular matrix (ECM) in the perisinusoidal space. In turn the three-dimensional structure of the ECM regulates the proliferation, morphology, and functions of the stellate cell. In this review, the morphology of sites of adhesion between hepatic stellate cells and extracellular matrix is described. Hepatic stellate cells cultured in polystyrene dishes spread well, whereas the cells cultured on or in type I collagen gel become slender and elongate their long cellular processes which adhere directly to the collagen fibers. Cells in type I collagen gel form a large number of adhesive structures, each adhesive area forming a face but not a point. Adhesion molecules, integrins, for the ECM are localized on the cell surface. Elongation of the cellular processes occurs via integrin-binding to type I collagen fibers. The signal transduction mechanism, including protein and phosphatidylinositol phosphorylation, is critical to induce and sustain the cellular processes. Information on the three-dimensional structures of ECM is transmitted via three-dimensional adhesive structures containing the integrins.Key words: hepatic stellate cells/extracellular matrix/cell-extracellular matrix adhesion/type I collagen/three-dimensional structureThe extracellular matrix (ECM) is known to regulate a diversity of biological activities, including cell proliferation, differentiation, migration, polarity, tumorigenesis and cancer metastasis. The cells adhere to ECM by specialized cellular devices such as focal contacts, podosomes, point contacts, and hemidesmosomes (Senoo and Hata, 1994b).The ECM is composed of an insoluble complex of collagens, adhesive glycoproteins (e.g., fibronectin, laminin), proteoglycans (PGs) (e.g., heparan sulfate PGs, chondroitin sulfate PGs) and elastin. These ECM components construct an insoluble supramolecular complex (reviewed by Hata, 1996), the molecules of which contain positional information relayed back as signals to the cells that produce them or to neighboring cells, as well as regulate their growth and metabolism (reviewed by Hata, 1996).Integrins are the major family of cell surface receptors that mediate attachment to the ECM (Hynes, 1992;Clark and Brugge, 1995;Giancotti and Ruoslahti, 1999). Formation of focal contacts is regulated by intracellular signals which promote integrin clustering and cytoskeletal associations. The relationship between the cells and ECM is one of the most important aspects with regard to the nature of the cells.Hepatic stellate cells (vitamin A-storing cells, lipocytes, fat-storing cells) lie in the perisinusoidal space (space of Disse) (Wake, 1980(Wake, , 1995(Wake, , 1997 and coexist with the threedimensional ECM components within the space. The types of ECM components regulate the behavior of the stellate cell (Senoo et al.,...

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Cellular interactions promote tissue-specific function, biomatrix deposition and junctional communication of primary cultured hepatocytes

1988

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Hepatocytes, prepared from normal adult rat liver, were seeded onto a collagen substratum and cultured alone or in the presence of rat liver endothelial cells. When hepatocytes were cultured alone in a hormonally defined serum-free medium, decreased albumin production and rapid morphological deterioration of bile canaliculi structures and gap junctions occurred within 4 to 5 days. In contrast, hepatocytes cocultured with liver mesenchymal cells remained morphologically intact and biochemically functional for at least 4 weeks. They reorganized into small islands, continued to secrete high levels of albumin, did not express alpha-fetoprotein (a fetal marker), and remained strongly dye coupled. All of the hepatocytes synthesized albumin and retained their gap junctional channels. No junctional communication was observed between hepatocytes and endothelial cells. Long fibers containing fibronectin, Type I collagen and laminin distributed over the hepatocytes were induced in coculture but never appeared in hepatocytes cultured alone. Moreover, supplementation of the hormonally defined medium with phenobarbital and dimethyl sulfoxide, both of which improve the life span and functional activities of cultured hepatocytes, failed to induce reticulin fiber formation in pure culture of hepatocytes. The modulation of albumin secretion, biomatrix deposition and junctional communication observed in hepatocytes cultured with sinusoidal liver cells was also obtained when hepatocytes were in association with various epithelial or mesenchymal cells [rat liver epithelial cells (T51B), mouse embryonic fibroblasts (NIH 3T3), human or rat dermal fibroblasts and bovine aorta endothelial cells (AG 4762)].

show abstract

Immunogold localization of procollagen III, fibronectin and heparan sulfate proteoglycan on ultrathin frozen sections of the normal rat liver

Cited by 66 publications

References 27 publications

The HepG2 Extracellular Matrix Contains Separate Heparinase- and Lipid-releasable Pools of ApoE

The HepG2 Extracellular Matrix Contains Separate Heparinase- and Lipid-releasable Pools of ApoE

Adhesion between Cells and Extracellular Matrix with Special Reference to Hepatic Stellate Cell Adhesion to Three-dimensional Collagen Fibers.

Cellular interactions promote tissue-specific function, biomatrix deposition and junctional communication of primary cultured hepatocytes

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