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Volumn 274, Issue 5291, 1996, Pages 1379-1383

Liver failure and defective hepatocyte regeneration in interleukin-6- deficient mice

Author keywords

[No Author keywords available]

Indexed keywords

CYCLINE; INTERLEUKIN 1; INTERLEUKIN 6; ONCOPROTEIN; TRANSCRIPTION FACTOR; TUMOR NECROSIS FACTOR ALPHA;

EID: 0029846756     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.274.5291.1379     Document Type: Article
Times cited : (1362)

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    • Hepatectomized animals were anesthetized and ventral laparotomy was performed. Normal liver was prepared by subjecting animals to laparotomy followed by perfusion as described (26). One hour before the remnant liver was harvested and fixed, animals were injected intraperitoneally with BrdU (50 mg/kg) (0.2% solution in PBS) [B. Schutte, M. M. J. Reynders, F. T. Bosman, G. H. Blijham, J. Histochem. Cytochem. 35, 1343 (1987)]. The portal vein was cannulated with a 22-gauge angiocatheter, the liver was flushed with PBS, and 4% paraformaldehyde (pH 7.2) (4°C) was then perfused for 10 min at a rate of 6 ml/min. The fixed liver was removed and cut into 5-mm slices with a razor blade and then fixed for 1 hour in 4% paraformaldehyde at 4°C. An automated tissue processor was used to embed liver slices with paraffin. Tissue sections (5 μm) were cut on a microtome and adhered to poly-L-lysine-coated glass slides. Staining of fixed tissue samples with an antibody to BrdU (Boehringer Mannheim) allows one to discern proliferating cells (brown, stained nuclei) from quiescent ones (clear, unstained nuclei). The immunohistochemical study was performed essentially as described [S. M. Hsu, L. Raine, H. Fanger, Am. J. Clin. Pathol. 75, 734 (1981); L. E. Greenbaum, D. E. Cressman, B. A. Haber, R. Taub, J. Clin. Invest. 96 (1995)]. StatWorks and Student's t test, respectively, were used for statistical analyses on animal liver weights and DNA synthesis.
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    • Hepatectomized animals were anesthetized and ventral laparotomy was performed. Normal liver was prepared by subjecting animals to laparotomy followed by perfusion as described (26). One hour before the remnant liver was harvested and fixed, animals were injected intraperitoneally with BrdU (50 mg/kg) (0.2% solution in PBS) [B. Schutte, M. M. J. Reynders, F. T. Bosman, G. H. Blijham, J. Histochem. Cytochem. 35, 1343 (1987)]. The portal vein was cannulated with a 22-gauge angiocatheter, the liver was flushed with PBS, and 4% paraformaldehyde (pH 7.2) (4°C) was then perfused for 10 min at a rate of 6 ml/min. The fixed liver was removed and cut into 5-mm slices with a razor blade and then fixed for 1 hour in 4% paraformaldehyde at 4°C. An automated tissue processor was used to embed liver slices with paraffin. Tissue sections (5 μm) were cut on a microtome and adhered to poly-L-lysine-coated glass slides. Staining of fixed tissue samples with an antibody to BrdU (Boehringer Mannheim) allows one to discern proliferating cells (brown, stained nuclei) from quiescent ones (clear, unstained nuclei). The immunohistochemical study was performed essentially as described [S. M. Hsu, L. Raine, H. Fanger, Am. J. Clin. Pathol. 75, 734 (1981); L. E. Greenbaum, D. E. Cressman, B. A. Haber, R. Taub, J. Clin. Invest. 96 (1995)]. StatWorks and Student's t test, respectively, were used for statistical analyses on animal liver weights and DNA synthesis.
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    • Hepatectomized animals were anesthetized and ventral laparotomy was performed. Normal liver was prepared by subjecting animals to laparotomy followed by perfusion as described (26). One hour before the remnant liver was harvested and fixed, animals were injected intraperitoneally with BrdU (50 mg/kg) (0.2% solution in PBS) [B. Schutte, M. M. J. Reynders, F. T. Bosman, G. H. Blijham, J. Histochem. Cytochem. 35, 1343 (1987)]. The portal vein was cannulated with a 22-gauge angiocatheter, the liver was flushed with PBS, and 4% paraformaldehyde (pH 7.2) (4°C) was then perfused for 10 min at a rate of 6 ml/min. The fixed liver was removed and cut into 5-mm slices with a razor blade and then fixed for 1 hour in 4% paraformaldehyde at 4°C. An automated tissue processor was used to embed liver slices with paraffin. Tissue sections (5 μm) were cut on a microtome and adhered to poly-L-lysine-coated glass slides. Staining of fixed tissue samples with an antibody to BrdU (Boehringer Mannheim) allows one to discern proliferating cells (brown, stained nuclei) from quiescent ones (clear, unstained nuclei). The immunohistochemical study was performed essentially as described [S. M. Hsu, L. Raine, H. Fanger, Am. J. Clin. Pathol. 75, 734 (1981); L. E. Greenbaum, D. E. Cressman, B. A. Haber, R. Taub, J. Clin. Invest. 96 (1995)]. StatWorks and Student's t test, respectively, were used for statistical analyses on animal liver weights and DNA synthesis.
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    • note
    • We thank J. Darnell for the STAT3 cDNA, U. Muller Eberhard and S. Maeda for the hemopexin (HPX) and serum amyloid P-component (SAP) probes, and C. Steer for the gift of the cyclin D1 cDNA and helpful discussions. We also thank C. Deutschmann, R. Diamond, D. Tewari, P. Traber, M. Lazar, and F. Rauscher for helpful discussions; S. Hwang and V. Miles for technical assistance; and J. Matthews for assistance with manuscript preparation. This work was in part supported by NIH grants DK44237, DK49210, and DK49629 (to R.T.); NIH grant K08 (to L.E.G.); the University of Pennsylvania Genetics Training Grant (to R.A.D.); and the Howard Hughes Medical Institute.


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