Summary
In the present study, we have localized by immunocytochemistry at the LM and EM level, procollagen type III (PIIIP), fibronectin (FN) and heparan sulfate proteoglycan (HSPG). Intracellularly, PIIIP was observed in both parenchymal and endothelial cells. In parenchymal cells, PIIIP was found in Golgi derived vesicles. This observation suggests that PIIIP synthesis is a normal function liver parenchymal cells. In endothelial cells, vesicles, which could not be identified, were seen to contain PIIIP. This result does not allow to conclude, whether sinusoidal endothelial cells secrete or take up PIIIP. Extracellularly, PIIIP was present around portal and central veins, in the space of Disse and between adjacent parenchymal cells. In the space of Disse, almost all interstitial collagen fibrils reacted with the anti PIIIP antibodies. This observation leads to the conclusion that most fibrils of the space of Disse contain type III in addition to type I collagen molecules.
By immunofluorescence, FN was seen mainly along the sinusoids in discrete dots. By EM, FN was found to be present in diffuse material closely associated with the sinusoidal membrane of the parenchymal cells and in strands connecting adjacent parenchymal cells, parenchymal and endothelial cells or parenchymal cells and collagen fibrils. FN was also present in vascular and ductular basal laminae.
Strong HSPG reaction was observed around bile ducts. Moderate reaction was seen around blood vessels and in the space of Disse. In the latter location, the ultrastructural distribution of HSPG resembles that of FN, i.e. HSPG is present in diffuse material and in strands.
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Beil N, Furthmayr H, Timpl R (1972) Chicken antibodies to soluble rat collagen, I. Characterization of the immune response by precipitation and agglutination methods. Immunochemistry 9:779–788
Bianchi FB, Biagini G, Ballardini G, Cenacchi G, Faccani A, Pisi E, Lasche R, Liotta L, Garbisa S (1984) Basement membrane production by hepatocytes in chronic liver disease. Hepatology 4:1167–1172
Biempica L, Morecky R, Wu CH, Giambione MA, Rojkind M (1980) Immunocytochemical localization of type B collagen. A component of basement membrane in human liver. Am J Pathol 98:591–602
Carlsson R, Engvall E, Freeman A, Ruoslahti E (1981) Laminin and fibronectin in cell adhesion: enhanced adhesion of cells from regenerating liver to laminin. Proc Natl Acad Sci USA 78:2403–2406
Clément B, Emonard H, Rissel M, Druguet M, Grimaud JA, Herbage D, Bourel M, Guillouzo A (1984) Cellular origin of collagen and fibronectin in the liver. Cell Mol Biol 30:489–496
De Leeuw AM, McCarthy S, Geerts A, Knook DL (1984) Isolated fat-storing cells divide and contain collagen. Hepatology 4:392–403
De Zanger R, Wisse E (1982) The filtration effect of rat liver fenestrated sinusoidal endothelium on the passage of (remnant) chylomicrons to the space of Disse. In: Knook DL, Wisse E (eds) Sinusoidal liver cells. Elsevier Biomedical, Amsterdam, pp 69–76
Gay S, Fietzek PP, Remberger K, Eder M, Kühn K (1975) Liver cirrhosis: immunofluorescence and biochemical studies demonstrate two types of collagen. Klin Wochemschr 53:205–208
Geerts A, Voss B, Rauterberg J, Wisse E (1982) On the distribution of collagen, fibronectin and laminin in the normal rat liver. In: Knook DL, Wisse E (eds) Sinusoidal liver cells. Elsevier Biomedical, Amsterdam, pp 209–216
Grimaud JA, Druguet M, Peyrol S, Chevalier O, Herbage D, El Badrawy N (1980) Collagen immunotyping in human liver: light and electron microscope study. J Histochem Cytochem 28:1145–1156
Hahn E, Wick G, Pencev D, Timpl R (1980) Distribution of basement membrane proteins in normal and fibrotic human liver: collagen IV, laminin and fibronectin. Gut 21:63–71
Hata R, Ninomiya Y, Nagai Y, Tsukada Y (1980) Biosynthesis of interstitial types of collagen by albumin-producing rat liver parenchymal cell (hepatocyte) clones in culture. Biochemistry 19:169–176
Hatahara T, Seyer JM (1982) Procollagen production by rat hepatocytes in primary culture (BBA 21134). Biochim Biophys Acta 716:431–438
Konomi H, Sano J, Nagai Y (1981) Immunohistochemical localization of type I, III and IV (Basement membrane) collagens in the liver. Acta Pathol Jpn 31:973–978
Martinez-Hernandez A (1984) The hepatic extracellular matrix, I. Electron immunohistochemical studies in normal rat liver. Lab Invest 51:57–75
Ninomiya Y, Hata R, Nagai Y (1980) Glycosaminoglycan synthesis by liver parenchymal cell clones in culture and its change with transformation. Biochim Biophys Acta 629:349–358
Orkin RW, Gehron P, McGoodwin EB, Martin GR, Valentine T, Swarm R (1977) A murine tumor producing a matrix of basement membrane. J Exp Med 145:204–220
Ponce P, Cordero J, Rojkind M (1981) A noncollagenous matrix for attachment of rat hepatocytes in culture. Hepatology 1:204–210
Popper H, Kent G (1975) Fibrosis in chronic liver disease. Clin Gastroenterol 4:315–332
Rauterberg J, Schlief H-E, Pott G (1982) The collagen of the normal and fibrotic liver. Characterization of collagen-derived peptides solubilized by cyanogenbromide cleavage and low temperature pepsin treatment. In: Gerlach U, Pott G, Rauterberg J, Voss B (eds) Connective tissue of the normal and fibrotic human liver. Georg Thieme, Stuttgart New York, pp 136–137
Rennard SI, Berg R, Martin GR, Foidart JM, Robey PG (1980) Enzyme-linked immunoassay (ELISA) for connective tissue components. Anal Biochem 104:205–214
Rojkind M, Giambrone MA, Biempica L (1979) Collagen types in normal and cirrhotic liver. Gastroenterology 76:710–719
Rojkind M, Kershenobich D (1983) The extracellular matrix: fibrosis and cirrhosis. In: Arias IM, Frenkel M, Wilson JHP (eds) The liver annual. Elsevier, Amsterdam New York Oxford, vol 3, pp 189–205
Sano J, Sato S, Ishizaki M, Yajima G, Konomi H, Fujiwara S, Nagai Y (1981) Types I, III and IV (basement membrane) collagens in the bovine liver parenchyma: electron microscopic localization by the peroxidase labeled antibody method. Biomed Res 2:546–551
Schulz R, Hahn U, Schuppan D, Hahn EG, Riecken EO (1984) Expression neuer Kollagen Typen durch portale Fibroblasten bei der obstructiven Gallengangserkrankung. Verh Dtsch Ges Inn Med 90:1499–1501
Schuppan D, Rühlmann T, Rebhuhn S, Hahn EG, Riecken EO (1984) A method that allows to quantify basement membrane collagen (type IV) and a new interstitial collagen (type VI) in liver biopsies (abstract). Gastroenterology 86:1339
Schuppan D, Dumont JM, Kim KY, Hennings G, Hahn EG (1986) Serum concentration of the aminoterminal procollagen type III peptide in the rat reflects early formation of connective tissue in experimental liver fibrosis. J. Hepatol (accepted for publication)
Seyer JM (1980) Interstitial collagen polymorphism in rat liver with CCl4-induced cirrhosis. Biochim Biophys Acta 629:490–498
Slot JW, Geuze HJ (1981) Sizing of protein A-colloidal gold probes for immunoelectron microscopy. J Cell Biol 90:533–536
Timpl R (1976) Immunological studies on collagen. In: Ramachandran GN, Reddi AH (eds) Biochemistry of collagen. Plenum Press, New York, pp 319–375
Timpl R (1982) Antibodies to collagens and procollagens. In: Cunningham LW, Frederiksen DW (eds) Methods in enzymology, vol 82, Structural and contractile proteins. (Part A) Extracellular matrix. Academic Press, New York London, pp 472–498
Tseng SCG, Smuckler EA, Stern R (1983) Types of collagen synthesized by normal rat liver hepatocytes in primary culture. Hepatology 3:955–963
Tokuyasu KT, Singer SJ (1976) Improved procedures for immunoferritin labeling of ultrathin frozen sections. J Cell Biol 71:894–906
Voss B, Allam S, Rauterberg J, Ullrich K, Gieselman V, von Figura K (1979) Primary cultures of rat hepatocytes synthesize fibronectin. Biochem Biophys Res Commun 90:1348–1354
Voss B, Rauterberg J, Allam S, Pott G (1980) Distribution of collagen type I and type III and of two collagenous components of basement membranes in the human liver. Pathol Res Pract 170:50–60
Wick G, Brunner H, Penner E, Timpl R (1978) The diagnostic application of specific antiprocollagen sera. II Analysis of liver biopsies. Int Arch Allerg Appl Immunol 56:316–324
Wisse E, De Wilde A, De Zanger R (1984) Perfusion fixation of human and rat liver tissue for light and electron microscopy: a review and assessment of existing methods with special emphasis on sinusoidal cells and microcirculation. In: SEM inc, O'Hare AMF (eds) Science of biological specimen preparation. Chicago IL USA, pp 31–38
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In honour of Prof. P. van Duijn
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Geerts, A., Geuze, H.J., Slot, J.W. et al. Immunogold localization of procollagen III, fibronectin and heparan sulfate proteoglycan on ultrathin frozen sections of the normal rat liver. Histochemistry 84, 355–362 (1986). https://doi.org/10.1007/BF00482963
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DOI: https://doi.org/10.1007/BF00482963