Loss of growth hormone-dependent characteristics of rat hepatocytes in culture

In Vitro Cellular & Developmental Biology - Plant - Tập 23 - Trang 303-307 - 1987
David W. Crabb1, Janet Roepke1
1Departments of Medicine and Biochemistry, Indiana University School of Medicine, Indianapolis

Tóm tắt

The liver of rodents is sexually differentiated, i.e. the female liver differs from the male liver. This differentiation is largely controlled by the pattern of growth hormone (GH) secretion. We have attempted to maintain GH-dependent differentiation of cultured rat hepatocytes. We examined the level of alcohol dehydrogenase (ADH) activity, which responds to GH and is higher in female than in male liver, and the estrogen receptor, which is dependent on GH but is present in equal amounts in males and females. ADH activity was maintained in cells from male rats, but fell by 40% in cells from females in medium supplemented with insulin and dexamethasone. The estrogen receptor content of female cells fell dramatically to undetectable levels within 2 d of culture. Extensive supplementation of the medium failed to prevent the decrease in ADH activity in female cells; similarly, the addition of female sex steroids; rat serum; pituitary extracts; rat, human, or bovine GH; or ovine prolactin failed to maintain the enzyme activity. Insulin, dexamethasone, thyroid hormone plus GH or prolactin, or the combination of all five hormones also failed to prevent the loss of estrogen receptors. Short-term cultures of rat hepatocytes, although retaining the liver-specific expression of ADH at the male level, lose GH-dependent expression of the estrogen receptor and stimulation of ADH activity.

Tài liệu tham khảo

Baxter, R. C.; Bryson, J. M.; Turtle, J. R. Somatogenic receptors of rat liver: regulation by insulin. Endocrinology 107:1176–1181; 1980. Baxter, R. C.; Zaltsman, Z.; Turtle, J. R. Rat growth hormone (GH) but not prolactin (PRL) induces both GH and PRL receptors in female rat liver. Endocrinology 114:1893–1901: 1984. Crabb, D. W.; Li, T.-K. Expression of alcohol dehydrogenase in primary monolayer cultures of rat hepatocytes. Biochem. Biophys. Res. Comm. 128:12–17; 1985. Crabb, D. W.; Morzorati, S.; Simon, J., et al. Central nervous system control of liver alcohol dehydrogenase activity in rats. Life Sci. 37:2381–2387; 1985. Crabb, D. W.; Bosron, W. F.; Li, T.-K. The role of the pituitary and androgenic imprinting in the regulation of liver alcohol dehydrogenase activity. Biochem. Pharmacol. 35:1527–1532; 1986. Eagon, P. K.; Fisher, S. E.; Imhoff, A. F., et al. Estrogen binding proteins of the male liver: influences of hormonal changes. Arch. Biochem. Biophys. 201:486–499; 1980. Eden, S. Age- and sex-related differences in episodic growth hormone secretion in the rat. Endocrinology 105:555–560; 1979. Enat, R.; Jefferson, D. M.; Ruiz-Opazo, N., et al. Hepatocyte proliferationin vitro: its dependence on the use of serum-free hormonally define medium and the substrate of extracellular matrix. Proc. Natl. Acad. Sci. USA 81:1411–1415; 1984. Gatmaitan, Z.; Jefferson, D. M.; Ruiz-Opazo, N., et al. Regulation of growth and differentiation of a rat hepatoma cell line by the synergistic interactions of hormones and the collagenous substrata. J. Cell Biol. 97:1179–1190; 1983. Gustafsson, J.-A.; Mode, A.; Norstedt, G., et al. Sex steroid-induced changes in hepatic enzymes. Ann. Rev. Physiol. 45:51–60; 1983. Gustafsson, J.-A.; Eden, S.; Eneroth, P., et al. Regulation of sexually dimorphic hepatic metabolism by the somatostatin-growth hormone axis. J. Steroid Biochem. 19:691–698; 1983. Gustafsson, J.-A.; Mode, A.; Norstedt, G. Central control of prolactin and estrogen receptors in rat liver. Expression of a novel endocrine system, the hypothalamo-pituitary-liver axis. Ann. Rev. Pharmacol. Toxicol. 23:259–278; 1983. Isom, H. C.; Georgoff, I. Quantitative assay for albumin-producing liver cells after simian virus 40 transformation of rat hepatocytes maintained in chemically defined medium. Proc. Natl. Acad. Sci. USA 81:6378–6382; 1984. Isom, H. C.; Secott, T.; Georgoff, I., et al. Maintenance of differentiated rat hepatocytes in primary culture. Proc. Natl. Acad. Sci. USA 82:3252–3256; 1985. Layne, E. Spectrophotometric and turbidometric methods for measuring proteins. Methods Enzymol. 3:447–454; 1957. Lin, R. C.; Snodgrass, P. J. Primary cultures of normal adult rat liver cells which maintain stable urea cycle enzymes. Biochem. Biophys. Res. Comm. 64:725–734; 1976. Mezey, E.; Potter, J. J. Rat liver alcohol dehydrogenase activity: effects of growth hormone and hypophysectomy. Endocrinology 104:1667–1673; 1979. Mezey, E.; Potter, J. J.; Tsitouras, P. D. Liver alcohol dehydrogenase activity in the female rat: effects of ovariectomy and estradiol administration. Life Sci. 29:1171–1176; 1981. Mode, A.; Norstedt, G.; Simic, B., et al. Continuous infusion of growth hormone feminizes hepatic steroid metabolism in the rat. Endocrinology 108:2103–2108; 1981. Norstedt, G.; Wrange, O.; Gustafsson, J.-A. Multihormonal regulation of the estrogen receptor in rat liver. Endocrinology 108:1190–1196; 1981. Norstedt, G.; Palmiter, R. Secretory rhythm of growth hormone regulates sexual differentiation of mouse liver. Cell 36:805–812; 1984. Posner, B. I.; Patel, B.; Vezinhet, A., et al. Pituitary-dependent growth hormone receptors in rabbit and sheep liver. Endocrinology 107:1954–1958; 1980. Rachamin, G.; MacDonald, J. A.; Wahid, S., et al. Modulation of alcohol dehydrogenase and ethanol metabolism by sex hormones in the spontaneously hypertensive rat. Biochem. J. 186:483–490; 1980. Scott, C. D.; Martin, J. L.; Baxter, R. C. Rat hepatocyte insulin-like growth factor I and binding protein: effect of growth hormone in vitro and in vivo. Endocrinology 116:1102–1107; 1985. Teschke, R.; Wannagat, F.-J.; Lowendorf, F., et al. Hepatic alcohol metabolizing enzymes after prolonged administration of sex hormones and alcohol in female rats. Biochem. Pharmacol. 35:521–527; 1986. Wakai, K.; Tsushia, T.; Isozaki, O., et al. Processing of human growth hormone by rat hepatocytes in monolayer culture. Endocrinology 114:1475–1482; 1984.