Three-dimensional examination of hepatic stellate cells in rat liver and response to endothelin-1 using confocal laser scanning microscopy
Corresponding Author
Hiroki Oikawa
Department of Pathology, School of Medicine, Iwate Medical University, Morioka, Japan
Dr H Oikawa, Department of Pathology, School of Medicine, Iwate Medical University, Uchimaru 19-1, Morioka 020-8505, Japan. Email: [email protected]Search for more papers by this authorTomoyuki Masuda
Department of Pathology, School of Medicine, Iwate Medical University, Morioka, Japan
Search for more papers by this authorJunzo Kawaguchi
Department of Pathology, School of Medicine, Iwate Medical University, Morioka, Japan
Search for more papers by this authorRyo Sato
Department of Pathology, School of Medicine, Iwate Medical University, Morioka, Japan
Search for more papers by this authorCorresponding Author
Hiroki Oikawa
Department of Pathology, School of Medicine, Iwate Medical University, Morioka, Japan
Dr H Oikawa, Department of Pathology, School of Medicine, Iwate Medical University, Uchimaru 19-1, Morioka 020-8505, Japan. Email: [email protected]Search for more papers by this authorTomoyuki Masuda
Department of Pathology, School of Medicine, Iwate Medical University, Morioka, Japan
Search for more papers by this authorJunzo Kawaguchi
Department of Pathology, School of Medicine, Iwate Medical University, Morioka, Japan
Search for more papers by this authorRyo Sato
Department of Pathology, School of Medicine, Iwate Medical University, Morioka, Japan
Search for more papers by this authorAbstract
Abstract Background and Aim: Hepatic stellate cells (HSC) are located in the space of Disse and are considered to participate in the regulation of sinusoidal flow. The contractility of quiescent HSC in normal liver has remained controversial, unlike activated HSC in injured liver. The aim of the present study was to examine the morphological changes in quiescent HSC in response to endothelin-1 (ET-1) perfusion.
Methods: Sections (50 µm thick) obtained from 15 normal rat livers with or without ET-1 perfusion (1 or 400 nmol/L) were stained immunohistochemically with antiglial fibrillary acidic protein (GFAP) antibody and then examined using confocal laser scanning microscopy. For examination of HSC, hepatic lobules were divided into three anatomic regions from the portal areas to the central veins. The length of HSC cytoplasmic processes and area of the sinusoids relative to the section area, excluding portal tracts and central veins, were measured.
Results: The GFAP-positive HSC were distributed relatively evenly in the hepatic lobules and those in region 2 (the area between periportal and pericentral areas) tended to have longer cytoplasmic processes. Perfusion of 1 or 400 nmol/L ET-1 for 25 min resulted in swelling of the cell bodies of GFAP-positive HSC and condensation of the intermediate filaments compared with those perfused with buffer only. Although narrowing of the sinusoidal lumen was observed in each region after perfusion with 400 nmol/L ET-1, there was no apparent shortening of the cytoplasmic processes of HSC. These findings were also confirmed quantitatively.
Conclusion: In the normal rat liver, quiescent HSC are not involved in the regulation of sinusoidal blood flow in response to ET-1.
References
- 1 Wake K. ‘Sternzellen’ in the liver: Perisinusoidal cells with special reference to storage of vitamin A. Am. J. Anat. 1971; 132: 429–62.
- 2 Hendrics HFJ, Verhoofstad WAMM, Brouwer A, De Leeuw AM, Knook DL. Perisinusoidal fat-storing cells are the main vitamin A storage sites in rat liver. Exp. Cell Res. 1985; 160: 138–49.
- 3
Ramadoli G.
The stellate cell (Ito-cell, fat storing cell, lipocyte, perisinusoidal cell) of the liver. New insights into pathophysiology of an intriguing cell.
Virchows Arch. B
1991; 61: 147–58.
10.1007/BF02890417 Google Scholar
- 4 Blomhoff R, Wake K. Perisunusoidal stellate cells of the liver: Important roles in retinol metabolism and fibrosis. FASEB J. 1991; 5: 271–7.
- 5 Yokoi Y, Namihisa T, Kuroda H et al.. Immunocytochemical detection of desmin in fat-storing cells (Ito cells). Hepatology 1984; 4: 709–14.
- 6 Niki T, De Bleser PJ, Wu G, Van den Berg K, Wisse E, Geerts A. Comparison of glial fibrillary acidic protein and desmin staining in normal and CCl4-induced fibrotic rat livers. Hepatology 1996; 23: 1538–45.
- 7 Neubauer K, Knittel T, Aurisch S, Fellmer P, Ramadori G. Glial fibrillary acidic protein: A cell type specific marker for Ito cells in vivo and in vitro. J. Hepatol. 1996; 24: 719–30.
- 8 Ballardini G, Groff P, De Giorgi LB, Schuppan D, Bianchi FB. Ito cell heterogeneity: Desmin-negative Ito cells in normal rat liver. Hepatology 1994; 19: 440–6.
- 9 Knittel T, Kobold D, Piscaglia F et al.. Localization of the liver myofibroblasts and hepatic stellate cells in normal and diseased rat livers: Distinct roles of (myo-) fibroblast subpopulations in hepatic tissue repair. Histochem. Cell Biol. 1999; 112: 387–401.
- 10 Cassiman D, Van Pelt J, De Vos R et al.. Synaptophysin: A novel marker for human and rat hepatic stellate cells. Am. J. Pathol. 1999; 155: 1831–9.
- 11 Gard AL, White FP, Dutton GR. Extra-neural glial fibrillary acidic protein (GFAP) immunoreactivity in perisinusoidal stellate cells of rat liver. J. Neuroimmunol. 1985; 8: 359–75.
- 12 Takahashi-Iwanaga H, Fujita T. Application of an NaOH maceration method to a scanning electron microscopic observation of Ito cells in the rat liver. Arch. Histol. Cytol. 1986; 49: 349–57.
- 13 Wake K, Sato T. Intralobular heterogeneity of perisinusoidal stellate cells in porcine liver. Cell Tissue Res. 1993; 273: 227–37.
- 14 Bhathal PS. Presence of modified fibroblasts in cirrhotic livers in man. Pathology 1972; 4: 139–44.
- 15 Bhathal PS, Grossman HJ. Reduction of the increased portal vascular resistance of the isolated perfused cirrhotic rat liver by vasodilators. J. Hepatol. 1985; 1: 325–37.
- 16 Elliot AJ, Vo LT, Grossman VL, Bhathal PS, Grossman HJ. Endothelin-induced vasoconstriction in isolated perfused liver preparations from normal and cirrhotic rats. J. Gastroenterol. Hepatol. 1997; 12: 314–18.
- 17 Kawada N, Klein H, Decker K. Eicosanoid-mediated contractility of hepatic stellate cells. Biochem. J. 1992; 285: 367–71.
- 18 Pinzani M, Failli P, Ruocco C et al.. Fat-storing cells as liver-specific pericytes. Spatial dynamics of agonist-stimulated intracellular calcium transients. J. Clin. Invest. 1992; 90: 642–6.
- 19 Sakamoto M, Ueno T, Kin M et al.. Ito cell contraction in response to endothelin-1 and substance P. Hepatology 1993; 18: 978–83.
- 20 Rockey DC, Chung JJ. Inducible nitric oxide synthase in rat hepatic lipocytes and the effect of nitric oxide on lipocyte contractility. J. Clin. Invest. 1995; 95: 1199–206.
- 21 Suematsu M, Goda N, Sano T et al.. Carbon monoxide: An endogenous modulator of sinusoidal tone in the perfused rat liver. J. Clin. Invest. 1995; 96: 2431–7.
- 22 Bauer M, Bauer I, Sonin NV et al.. Functional significance of endothelin B receptors in mediating sinusoidal and extrasinusoidal effects of endothelins in the intact rat liver. Hepatology 2000; 31: 937–47.
- 23 Yanagisawa M, Kurihara H, Kimura S et al.. A novel potent vasoconstrictor peptide produced by vascular endothelial cells. Nature 1988; 332: 411–15.
- 24 Arai H, Hori S, Aramori I, Ohkubo H, Nakanishi S. Cloning and expression of a cDNA encoding an endothelin receptor. Nature 1990; 348: 730–2.
- 25 Sakurai T, Yanagisawa M, Takuwa Y et al.. Cloning of a cDNA encoding a non-isopeptide-selective subtype of the endothelin receptor. Nature 1990; 348: 732–5.
- 26 Housset C, Rockey DC, Bissell DM. Endothelin receptors in rat liver: Lipocytes as a contractile target for endothelin 1. Proc. Natl Acad. Sci. USA 1993; 90: 9266–70.
- 27 Housset CN, Rockey DC, Friedman SL, Bissell DM. Hepatic lipocytes: A major target for endothelin-1. J. Hepatol. 1995; 22 (Suppl. 2): 55–60.
- 28 Kawada N, Tran-Thi TA, Klein H, Decker K. The contraction of hepatic stellate (Ito) cells stimulated with vasoactive substances. Eur. J. Biochem. 1993; 213: 815–23.
- 29 Bauer M, Paquette NC, Zhang JX et al.. Chronic ethanol consumption increases hepatic sinusoidal contractile response to endothelin-1 in the rat. Hepatology 1995; 22: 1565–76.
- 30 Rockey DC, Housset CN, Friedman SL. Activation-dependent contractility of rat hepatic lipocytes in culture and in vivo. J. Clin. Invest. 1993; 92: 1795–804.
- 31 Rockey DC, Weisiger R. Endothelin induced contractility of stellate cells from normal and cirrhotic rat liver: Implications for regulation of portal pressure and resistance. Hepatology 1996; 24: 233–40.
- 32 Zhang JX, Pegoli Jr W, Clemens MG. Endothelin-1 induces direct constriction of hepatic sinusoids. Am. J. Physiol. 1994; 266: G624–G632.
- 33 Okumura S, Takei Y, Kawano S et al.. Vasoactive effect of endothelin-1 on rat liver in vivo. Hepatology 1994; 19: 155–61.
- 34 Housset C. The dual play of endothelin receptors in hepatic vasoregulation. Hepatology 2000; 31: 1025–6.
- 35
McCuskey RS.
Morphological mechanism for regulating blood flow through hepatic sinusoids.
Liver
2000; 20: 3–7.
10.1034/j.1600-0676.2000.020001003.x Google Scholar
- 36 Rockey DC. The cellular pathogenesis of portal hypertension: Stellate cell contractility, endothelin, and nitric oxide. Hepatology 1997; 25: 2–5.
- 37 Rockey DC. Hepatic blood flow regulation by stellate cells in normal and injured liver. Semin. Liver Dis. 2001; 21: 337–49.
- 38 Geerts A. History, heterogeneity, developmental biology, and functions of quiescent hepatic stellate cells. Semin. Liver Dis. 2001; 21: 311–35.DOI: 10.1055/s-2001-17550
- 39 Shay J. Economy of effort in electron microscope morphometry. Am. J. Pathol. 1975; 81: 503–12.
- 40 Beier K, Fahimi HD. Application of automatic image analysis for morphometric studies of peroxisomes stained cytochemically for catalase. Cell Tissue Res. 1986; 246: 635–40.
- 41 Wisse E, De Zanger RB, Charels K, Van Der Smissen P, McCuskey RS. The liver sieve: Considerations concerning the structure and function of the endothelial fenestrae, the sinusoidal wall and the space of Disse. Hepatology 1985; 5: 683–92.
- 42 Motta P, Porter KR. Structure of rat liver sinusoids and associated tissue spaces as revealed by scanning electron microscopy. Cell Tissue Res. 1974; 148: 111–25.
- 43 Lautt WW, Greenway CV, Legare DJ, Weisman H. Localization of intrahepatic portal vascular resistance. Am. J. Physiol. 1986; 251: G375–81.
- 44 Zhang JX, Bauer M, Clemens MG. Vessel- and target cell-specific actions of endothelin-1 and endothelin-3 in rat liver. Am. J. Physiol. 1995; 269: G269–77.
- 45 Gondo K, Ueno T, Sakamoto M, Sakisaka S, Sata M, Tanikawa K. The endothelin-1 binding site in rat liver tissue. Light- and electron-microscopic autoradiographic studies. Gastroenterology 1993; 104: 1745–9.
- 46 Wake K. Sinusoidal structure and dynamics. In: Vidal-Vanaclocha F, ed. Functional Heterogeneity of Liver Tissue: From Cell Lineage Diversity to Sublobular Compartment-Specific Pathogenesis. Austin, Texas: RG Landes, 1996; 57–67.
- 47 Kristensen DB, Kawada N, Imamura K et al.. Proteome analysis of rat hepatic stellate cells. Hepatology 2000; 32: 268–77.
- 48 Kaneda K, Ekataksin W, Sogawa M, Matsumura A, Cho A, Kawada N. Endothelin-1-induced vasoconstriction causes a significant increase in portal pressure of rat liver: Localized constrictive effect on the distal segment of preterminal portal venules as revealed by light and electron microscopy and serial reconstruction. Hepatology 1998; 27: 735–47.
- 49 McCuskey RS, Ito Y, McCuskey MK, Ekataksin W, Wake K. Morphologic mechanisms for regulating blood flow through hepatic sinusoids: 1998 update and overview. In: Wisse E, Knook DL, DeZanger R, Fraser R, eds. Cells of the Hepatic Sinusoid, Vol. 7. Leiden, The Netherlands: Kupffer Cell Foundation, 1999; 129–34.
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