Abstract
In order to study the influence of cell shape as modulated by the extracellular matrix on the cellular activity, hepatocytes isolated from liver were maintained on collagen I coated plastic substrata and collage I gel substrata and certain hepatocyte specific functions were investigated. The incorporation of3[H]-leucine into total proteins and albumin secreted by cells maintained on collagen gel was found to be significantly higher compared to those maintained on a collagen coated plastic substrata, indicating that hepatocytes on collagen gel have an enhanced albumin synthesizing capacity. Increased incorporation of35[S]-sulphate into total proteoglycans (PG) and a relatively higher fraction of the35[S]-PG in the extracellular space showed an increased rate of synthesis and secretion of sulphated PGs by cells maintained on collagen gels. But in contrast to the above results, the incorporation of3[H]-leucine into cytokeratins C8, C18 and actin were significantly low in cells maintained on collagen gel. The tyrosine amino transferase activity exhibited by hepatocytes preincubated with dexamethasone on collagen gel was also significantly low. The different forms of collagen substrata appeared to have no effect on the amino acid transport by hepatocytes, further suggesting that the various hepatocyte specific functions are not uniformly altered when hepatocytes are maintained on three-dimensional collagen gel substrata. These results indicate that the shape of the cell as determined by the nature of the matrix substratum influences the synthetic activity of secretory proteins and those remaining intracellularly, differently.
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References
Hay ED: Extracellular matrix. J Cell Biol 91: 205s-223s, 1981
Montesano R, Mouron P, Amherdt M, Orci L: Collagen matrix promotes reorganisation of pancreatic endocrine cell monolayers into islet like organoids. J Cell Biol 97: 935–939, 1983
Chen J, Stuckey EC, Berry CL: Three dimensional culture of rat exocrine pancreatic cells using collagen gels. Br J Exp Pathol 66: 551–559, 1985
Sudhakaran PR, Stamatoglou SC, Hughes RC: Modulation of protein synthesis and secretion by substratum in primary cultures of Rat hepatocytes. Exp Cell Res 167: 505–516, 1986
Rojkind M, Gatmaitan Z, Mackensen S, Giambrone M, Ponce P, Reid LM: Connective tissue biomatrix: Its isolation and utilization for long term cultures of normal rat hepatocytes. J Cell Biol 87: 255–263, 1980
Bissell DM, Arenson DM, Maher JJ, Roll FJ: Support of cultured hepatocytes on a Laminin rich gel. J Clin Invest 79: 801–812, 1987
Ben-Zeev A, Robinson GS, Bucher NLR, Farmer SR: Cell-cell and cell-matrix interactions differentially regulate the expression of hepatic and cytoskeletal genes in primary cultures of rat hepatocytes. Proc Natl Acad Sci, USA 85: 2161–2165, 1988
Nakamura T, Teramoto H, Tomita Y, Ichihara A: L-proline is an essential amino acid for hepatocyte growth in culture. Biochem Biophys Res Commun 122: 884–891, 1984
Buck CA, Horwitz AF: Cell surface receptors for extracellular matrix molecules. Ann Rev Cell Biol 3: 179–205, 1987
Albelda SM, Buck CA: Integrins and other cell adhesion molecules. FASEB J 4: 2868–2880, 1990
Gull-berg D, Terracio L, Borg TK, Rubin K: Identification of Integrin like matrix receptors with affinity for Interstitial collagens. J Biol Chem 264: 12686–12694: 1989
Ruoslahti E: Fibronectin and its receptors. Ann Rev Biochem 57: 375–413, 1988
Forseberg E, Paulsson M, Timpl R, Johansson S: Characterization of a laminin receptor on rat hepatocytes. J Biol Chem 265: 6376–6381, 1990
Ben-Zeev A: The relation between cytoplasmic organisation, gene expression and morphogenesis. Trends Biochem Sci 11: 478–481, 1986
Spooner BS: Microfilaments, microtubules and extracellular materials in morphogenesis. Bioscience 25: 440–451, 1975
Gospodarowicz D, Greenburg G, Birdwell CR: Determination of cellular shape by the extracellular matrix and its correlation with control of cellular growth. Cancer Res 38: 4155–4171, 1978
Emerman JT, Pitelka DR: Maintenance and induction of morphological differentiation in dissociated mammary epithelium on floating collagen membranes.In vitro 13: 316–328, 1977
Seglen PO: Preparation of isolated rat liver cells. Methods Cell Biol 13: 29–83, 1976
Sudhakaran PR, Sinn W, von Figura K: Regulation of heparan sulphate metabolism by adenosine 3′5′ cyclic monophosphate in hepatocytes in culture. Biochem J 192: 395–402, 1980
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275, 1951
Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685, 1970
Chamberlain JB: Fluorographic detection of radioactivity in polyacrylamide gels with the water soluble fluor, sodium salicylate. Anal Biochem 98: 132–135, 1979
Granner DK, Tomkins GM: Tyrosine amino transferase (Rat liver). Methods Enzymol 17A: 633–637, 1970
Sudhakaran PR, Sinn W, von Figura K: Initiation of altered heparan sulphate on β-D-xyloside in rat hepatocytes. Hoppe-Seylers Z Physiol. Chem 362: 39–46, 1981
Bennett CA, Franklin NL: In: Statistical analysis in Chemistry and Chemical Industry, John Wiley and sons, Inc, New York
Li ML, Aggeler J, Farson DA, Hatier C, Hassell J, Bissell MJ: Influence of a reconstituted basement membrane and its components on casein gene expression and secretion in mouse mammary epithelial cells. Proc Natl Acad Sci USA 84: 136–140, 1987
Wicha MS, Lawrie G, Kohn E, Bagavandon P, Mahn T: Extracellular matrix promotes mammary epithelial growth on the differentiationin vitro. Proc Natl Acad Sci USA 79: 3213–3217, 1982
Schuetz EG, Li D, Omiecinski CJ, Muller-Eberhard U, Kleinman HK, Elswick B, Guzelian PS: Regulation of gene expression in adult rat hepatocytes cultured on a basement membrane matrix. J Cell Physiol 134: 309–323, 1988
Dunn JCY, Tompkins RG, Yarmush LM: Hepatocytes in collagen sandwich: Evidence for transcriptional and translational regulation. J Cell Biol 116: 1043–1053, 1992
Walling JM, Blackmore M, Hickman JA, Townsend KM: Role of the extracellular matrix on the growth and differentiated phenotype of murine colonic adenocarcinoma cellsin vitro. Int J Cancer 47: 776–783, 1991
Parry G, Lee EYH, Farson DA, Koval N, Bissell MJ: Collageneous substrate regulate the nature and distribution of glycosaminoglycans produced by differentiated cultures of mouse mammary epithelial cells. Exp Cell Res 156: 487–499, 1985
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Santhosh, A., Sudhakaran, P.R. Influence of collagen gel substrata on certain biochemical activities of hepatocytes in primary culture. Mol Cell Biochem 137, 127–133 (1994). https://doi.org/10.1007/BF00944074
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DOI: https://doi.org/10.1007/BF00944074