Summary
To study the process of the expression of reduced nicotinamide adenine dinucleotide phosphate-cytochrome P-450 reductase (EC 1.6.2.4) in the liver during development, the amount of enzyme in the cytoplasm of periportal and perivenular hepatocytes in sections cut from livers of male rats was measured during peri- and postnatal growth by quantitative immunohistochemistry with a video image processor. In livers of 19-day-old foetuses, the reductase content in the cytoplasm of periportal and perivenular hepatocytes was 0.16 μM and 0.20 μM, respectively. From the 19th day of gestation to 5 days after birth, the enzyme content increased markedly in the cytoplasm of periportal (288%) and perivenular hepatocytes (301%). Subsequently, the content in the cytoplasm of periportal hepatocytes increased slightly (46%) from 5 to 20 days of age, remained unchanged from 20 to 45 days of age, and increased slightly (15%) from 45 to 90 days of age. However, the content in the cytoplasm of perivenular hepatocytes increased progressively (125%) between 5 and 90 days of age. Thus, the amount of cytochrome P-450 reductase increases markedly in periportal and perivenular hepatocytes during the perinatal period, and subsequently the enzyme content increases gradually in periportal hepatocytes and progressively in perivenular hepatocytes. The present results also suggest that the divergence between cytochrome P-450 expression and the cytochrome P-450-dependent drug metabolic activity in hepatocytes during the perinatal period, found in previous studies, can be attributed to a low cytochrome P-450 reductase density in the membrane of endoplasmic reticulum of periportal and perivenular hepatocytes.
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French, J. S., Guengerich, F. P. &Coon, M. J. (1980) Interaction of cytochrome P-450, NADPH-cytochrome P-450 reductase, phospholipid, and substrate in the reconstituted liver microsomal enzyme system.J. Biol. Chem. 255, 4112–19.
Gustafsson, J-A., Mode, A., Norstedt, G. &Skett, P. (1983) Sex steroid induced changes in hepatic enzymes.Ann. Rev. Physiol. 45, 51–60.
Herzfeld, A., Federman, M. &Greengard, O. (1973) Subcellular morphometric and biochemical analysis of developing rat hepatocytes.J. Cell. Biol. 57, 475–83.
Jacobson, M. &Kuntzman, R. (1969) Testosterone hydroxylation and pentobarbital oxidation by rat liver microsomes; selective effects of age, sex, castration and testosterone propionate treatment.Steroids 13, 327–41.
Kanai, K., Watanabe, J. &Kanamura, S. (1990) Microphotometric analysis of cytochrome P-450 in periportal, midzonal and perivenular hepatocytes of mice treated with phenobarbital.J. Histochem. Cytochem. 38, 1599–1605.
Kanai, K., Watanabe, J., Asaka, Y., Fujimoto, S. &Kanamura, S. (1992) Postnatal changes in sublobular distribution of NADPH-cytochrome P-450 reductase in rat liver.Histochem. J. 24, 957–63.
Lamers, W. H., Hilberts, A., Furt, E., Smith, J., Jonges, G. N., van Noorden, C. J. F., Gaasbeek-Janzen, J. W., Charles, R. &Moorman, A. F. M. (1989) Hepatic enzymic zonation: a reevaluation of the concept of the liver acinus.Hepatology 10, 72–6.
Maeda, K., Kamataki, T., Nagai, T. &Kato, R. (1984) Postnatal development of constitutive forms of cytochrome P-450 in liver microsomes of male and female rats.Biochem. Pharmacol. 33, 509–12.
Masaki, R., Yamamoto, A. &Tashiro, Y. (1987) Cytochrome P-450 and NADPH- cytochrome P-450 reductase are degraded in the autolysosomes in rat liver.J. Cell. Biol. 104, 1207–15.
Masters, B. S. S., Prough, R. A. &Kamin, H. (1975): Properties of the stable aerobic and anaerobic half reduced states of NADPH-cytochrome c reductase.Biochemistry 14, 607–13.
Miwa, G. T., West, S. B. &Lu, A. Y. H. (1978) Studies on the rate-limiting enzyme component in the microsomal monooxygenase system.J. Biol. Chem. 253, 1921–9.
Murray, G. I., Barnes, T. S., Sewell, H. F., Ewen, S. W. B., Melvin, W. T., Shaw, P. M., Fowler, J. &Burke, M. D. (1987) Cytochrome P-450 localization in normal human adult and foetal liver by immunocytochemistry using a monoclonal antibody against human cytochrome P-450.Histochem. J. 19, 537–45.
Omiecinski, C. J., Hassett, C. &Costa, P. (1990) Developmental expression and in situ localization of the phenobarbital-inducible rat hepatic mRNAs for cytochromes CYP2B1, CYP2B2, CYP2C6, and CYP3A1.Mol. Pharmacol. 38, 462–70.
Pool, C. F., Madlener, S., Diegenbach, P. C., Sluiter, A. A. &Van Dersluis, P. (1984) Quantitation of antiserum reactivity in immunocytochemistry. Two new methods for measuring peroxidase activity on antigen-coupled beads incubated according to an immunoperoxidase method.J. Histochem. Cytochem. 32, 921–8.
Ram, P. A. &Waxman, D. J. (1992) Thyroid hormone stimulation of NADPH P450 reductase expression in liver and extrahepatic tissues. Regulation by multiple mechanisms.J. Biol. Chem. 267, 3294–301.
Ratanasavanh, D., Beaune, P., Morel, F., Flinois, J.-P., Guengerich, F. P. &Guillouzo, A. (1991) Intralobular distribution and quantitation of cytochrome P-450 enzymes in human liver as a function of age.Hepatology 13, 1142–51.
Shapiro, B. H., Pampori, N. A., Lapenson, D. P. &Waxman, D. J. (1994) Growth hormone-dependent and-independent sexually dimorphic regulation of phenobarbital-induced hepatic cytochromes P-450 2B1 and 2B2.Arch. Biochem. Biophys. 312, 234–9.
Smith, M. T., Redick, J. A. &Baron, J. (1983) Quantitative immuno-histochemistry: a comparison of microdensitometric analysis of unlabeled antibody peroxidase-antiperoxidase staining and of microfluorometric analysis of indirect fluorescent antibody staining for nicotinamide adenosine dinucleotide phosphate (NADPH)-cytochrome (P-450) reductase in rat liver.J. Histochem. Cytochem. 31, 1183–9.
Taira, Y., Redick, J. A. &Baron, J. (1980) An immunohistochemical study on the localization and distribution of NADPH-cytochrome c (P-450) reductase in rat liver.Mol. Pharmacol. 17, 374–81.
Watanabe, J. &Kanamura, S. (1991) An improved microphotometry system for measurement of cytochrome P-450 in hepatocyte cytoplasm.J. Histochem. Cytochem. 39, 689–94.
Watanabe, J., Kanai, K. &Kanamura, S. (1991) Measurement of NADPH-ferrihemoprotein reductase content in sections of liver.J. Histochem. Cytochem. 39, 1635–43.
Watanabe, J., Asaka, Y., Kanai, K. &Kanamura, S. (1992) Relation between cytochrome P-450 increase and endoplasmic reticulum proliferation in hepatocytes of mice treated with phenobarbital: a microphotometric and morphometric study.J. Histochem. Cytochem. 40, 353–7.
Watanabe, J., Asaka, Y., Fujimoto, S. &Kanamura, S. (1993a) Densities of NADPH-ferrihemoprotein reductase and cytochrome P-450 molecules in the endoplasmic reticulum membrane of rat hepatocytes.J. Histochem. Cytochem. 41, 43–49.
Watanabe, J., Asaka, Y. &Kanamura, S. (1993b) Postnatal development and sublobular distribution of cytochrome P-450 in rat liver: a microphotometric study.J. Histochem. Cytochem. 41, 397–400.
Watanabe, J., Asaka, Y., Tanaka, T. &Kanamura, S. (1994) Measurement of NADPH-cytochrome P-450 reductase content in rat liver sections by quantitative immunohistochemistry with a video image processor.J. Histochem. Cytochem. 42, 1161–7.
Waxman, D. J., Dannan, G. A. &Guengerich, F. P. (1985) Regulation of rat hepatic cytochrome P-450: age-dependent expression, hormonal imprinting, and xenobiotic inducibility of sex-specific isoenzymes.Biochemistry 24, 4409–17.
Wolf, C. R., Moll, E., Friedberg, T., Oesch, F., Buchmann, A., Kuhlmann, W. D. &Kunz, H. W. (1984) Characterization, localization and regulation of a novel phenobarbital-inducible form of cytochrome P450, compared with three further P450-isozymes, NADPH P450-reductase, glutathione transferases and microsomal epoxide hydrolase.Carcinogenesis 5, 993–1001.
Yasukochi, Y. &Masters, B. S. S. (1976) Some properties of a detergent-solubilized NADPH-cytochrome c (P-450) reductase purified by biospecific affinity chromatography.J. Biol. Chem. 251, 5337–44.
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Watanabe, J., Asaka, Y. & Kanamura, S. Peri- and postnatal changes in reduced nicotinamide adenine dinucleotide phosphate-cytochrome P-450 reductase content in hepatocytes of rats. Histochem J 28, 505–510 (1996). https://doi.org/10.1007/BF02331410
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DOI: https://doi.org/10.1007/BF02331410