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NADPH:cytochrome P-450(c) reductase: Biochemical characterization in rat brain and cultured neurons and evolution of activity during development

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Abstract

NADPH:cytochrome P-450 (c) reductase is a microsomal enzyme which is involved in the cytochrome P-450-dependent biotransformation of many exogenous agents as well as of some endogenous molecules. Using cytochromec as a substrate, the kinetic parameters of this enzyme were determined in brain microsomes. The comparison of the NADPH:cytochrome P-450 reductase's Vmax values and cytochrome P-450 contents in both fractions, suggests a role of cerebral NADPH:cytochrome P-450 reductase in cytochrome P-450 independent pathways. This is also supported by the different developmental pattern of brain enzyme as compared to the liver enzyme, and by the presence of a relatively high NADPH:cytochrome P-450 reductase activity in immature rat brain and neuronal cultures, while cytochrome P-450 was hardly detectable in these preparations. The enzyme activity was not induced by a phenobarbital chronic treatment neither in the adult brain nor in cultured neurons, suggesting a different regulation of the brain enzyme expression.

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References

  1. Lu, A. Y. H., and West, S. B. 1980. Multiplicity of mammalian microsomal cytochrome P-450. Pharmacol. Rev. 31:277–295.

    Google Scholar 

  2. Masters, B. S. S. 1980. The role of NADPH-cytochromec (P-450) reductase in detoxication. Pages 183–200in Jakoby, W. B. (ed.) Enzymatic Basis of Detoxication. Vol. 1, Academic Press, New York.

    Google Scholar 

  3. Miwa, G. T., and Lu, A. Y. H. 1984. The association of cytochrome P-450 reductase in phospholipid membranes. Arch. Biochem. Biophys. 234:161–166.

    PubMed  Google Scholar 

  4. Gram, T. E., and Gilette, J. R. 1971. Biotransformation of drugs. Pages 571–609,in Bacq, Z. M. (ed.) Fundamentals of Biochemical Pharmacology, Pergamon Press, Oxford.

    Google Scholar 

  5. Holtzman, J. L., Crankshaw, D. L., Peterson, F. J., and Polnaszek, C. F. 1981. The kinetics of the aerobic reduction of nitrofurantoin by NADPH-cytochrome P-450 (c) reductase. Mol. Pharmacol. 20:669–673.

    PubMed  Google Scholar 

  6. Minchin, R. F., Ho, P. C., and Boyd, M. R. 1986. Reductive metabolism of nitrofurantoin by rat lung and liver in vitro. Biochem. Pharmacol. 35:575–580.

    PubMed  Google Scholar 

  7. Heimbrook, D. C., and Sartorelli, A. C. 1986. Biochemistry of misonidazole reduction by NADPH-cytochrome c (P-450) reductase. Mol. Pharmacol. 29:168–172.

    PubMed  Google Scholar 

  8. Sasame, H. A., Ames, M. M., and Nelson, S. D. 1977. Cytochrome P-450 and NADPH cytochrome c reductase in rat brain. Formation of catechols and reactive catechol metabolites. Biochem. Biophys. Res. Comm. 78:919–926.

    PubMed  Google Scholar 

  9. Nabeshima, T., Fontenot, J., and Ho, I. K. 1977. Effects of chronic administration of pentobarbital and morphine on the brain microsomal cytochrome P-450 system. Biochem. Pharmacol. 30:1142–1145.

    Google Scholar 

  10. Walther, B., Ghersi-Egea, J. F., Minn, A., and Siest, G. 1986. Subcellular distribution of cytochrome P-450 in the brain. Brain Res. 375:338–344.

    PubMed  Google Scholar 

  11. Paul, S. M., Axelrod, J., and Diliberto, E. J. 1977. Catechol oestrogens forming enzyme of brain—Demonstration of cytochrome P-450 monooxygenase. Endocrinology 101:1604–1610.

    PubMed  Google Scholar 

  12. Walther, B., Ghersi-Egea, J. F., Minn, A., and Siest, G. 1987a. Brain mitochondrial cytochrome P-450scc: spectral and catalytic properties. Arch. Biochem. Biophys. 254:592–596.

    PubMed  Google Scholar 

  13. Mesnil, M., Testa, B., and Jenner, P. 1985. Aryl hydrocarbon hydroxylase in rat brain microsomes. Biochem. Pharmacol. 34:435–436.

    Google Scholar 

  14. Walther, B., Ghersi-Egea, J. F., Jayyosi, Z., Minn, A., and Siest, G. 1987b. Ethoxyresorufin O-deethylase activity in rat brain subcellular fractions. Neurosci. Lett. 76:58–62.

    PubMed  Google Scholar 

  15. Possmayer, F., Kleine, L., Duwe, G., Steward-DeHaan, P. J., Wong, T., MacPherson, C. F. C., and Harding, P. G. R. 1979. Differences in the subcellular and subsynaptosomal distribution of the putative endoplasmic reticulum markers, NADPH-cytochromec reductase, estrone sulfate sulfohydrolase and CDP-choline-diacylglycerol cholinephosphotransferase in rat brain. J. Neurochem. 32:889–906.

    PubMed  Google Scholar 

  16. Takeshita, M., Miki, M., and Yubishi, T. 1982. Cytochrome reductase activities in rat brain during development. J. Neurochem. 39:1047–1049.

    PubMed  Google Scholar 

  17. Haglund, L., Kohler, C., Haaparenta, Y., Goldstein, M., and Gustafsson, J. A. 1984. Presence of NADPH-cytochrome P-450 reductase in central catecholaminergic neurones. Nature 307:259–262.

    PubMed  Google Scholar 

  18. Daval, J. L., Pereira de Vasconcelos, A., and Lartaud, I. 1988. Morphological and neurochemical influence of diazepam and phenobarbital on selective cultures of neurons from fetal rat brain. J. Neurochem. 50:665–672.

    PubMed  Google Scholar 

  19. Bottenstein, J. F., and Sato, G. 1979. Growth of a neuroblastoma cell line in serum-free supplemented medium. Proc. Nat. Acad. Sci. U.S. 76:514–517.

    Google Scholar 

  20. Faivre-Baumann, N., Rosenblum, E., Ruymirat, S., Grouselle, D., and Tixier-Vidal, A. 1981. Differentiation of fetal mouse hypothalamic cells in serum-free medium. Dev. Neurosci. 4:118–129.

    PubMed  Google Scholar 

  21. Ghersi-Egea, J. F., Walther, B., Minn, A., and Siest, G. 1987. Quantitative measurement of cerebral cytochrome P-450 by second derivative spectrophotometry. J. Neurosci. Meth. 20:261–269.

    Google Scholar 

  22. Beaufay, H., Amar-Costesec, A., Feymans, E., Thines-Sempoux, D., Wibo, M., Robbi, M., and Berther, J. 1974. Analytical study of microsomes and isolated subcellular membranes from rat liver. J. Cell Biol. 61:213–231.

    Google Scholar 

  23. Strobel, H. W., and Dignam, J. D. 1978. Purification and properties of NADPH-cytochrome P-450 reductase. Pages 89–93, in Fleischer, S. E. and Packer, I. (eds.) Methods in Enzymology, Vol 52, Academic Press, London.

    Google Scholar 

  24. Lowry, O. H., Rosenbrough, N. J., Farr, A. L., and Randall, R. J. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193:265–275.

    PubMed  Google Scholar 

  25. Dignam, J. D., and Strobel, H. W. 1977. Preparation of homogeneous NADPH-cytochrome P-450 reductase from rat liver. Biochem. Biophys. Res. Comm. 63:845–852.

    Google Scholar 

  26. Yasukochi, Y., and Masters, B. S. S. 1976. Some properties of detergent-solubilized NADPH-cytochromec (cytochrome P-450) reductase purified by biospecific affinity chromatography. J. Biol. Chem. 251:5337–5344.

    PubMed  Google Scholar 

  27. Souhaili-El Amri, H., Fargetton, X., Delatour, P., and Batt, A. M. 1987. Sulphoxidation of albendazole by the FAD-containing and cytochrome P-450-dependent monooxygenases from pig liver microsomes. Xenobiotica 17:1159–1168.

    PubMed  Google Scholar 

  28. Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685.

    PubMed  Google Scholar 

  29. Burnette, W. N. 1981. Western blotting: electrophoretic transfer of proteins from sodium dodecyl sufate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal. Biochem. 112:195–203.

    PubMed  Google Scholar 

  30. Guenthner, T. M., and Mannering, G. J. 1977. Induction of hepatic monooxygenase system in fetal and neonatal rats with phenobarbital, polycyclic hydrocarbons and other xenobiotics. Biochem. Pharmacol. 26:567–575.

    PubMed  Google Scholar 

  31. Cresteil, T., Flinois, J. P., Pfister, A., and Leroux, J. P. 1979. Effect of microsomal preparations and induction on cytochrome P-450-dependent monooxygenases in fetal and neonatal rat liver. Biochem. Pharmacol. 28:2057–2063.

    PubMed  Google Scholar 

  32. Conney, A. H. 1967. Pharmacological implications of microsomal enzyme induction. Pharmac. Rev. 19:317–366.

    Google Scholar 

  33. Noguchi, M., Yoshida, T., and Kikuchi, G. 1980. Specific requirement of NADPH-cytochrome c reductase for the microsomal heme oxygenase reaction yielding biliverdin IXa. FEBS Lett. 98:281–284.

    Google Scholar 

  34. Dailey, H. A., and Strittmatter, P. 1980. Characterization of the interaction of amphipathic cytochrome b5 with stearyl coenzyme A desaturase and NADPH:cytochrome P-450 reductase. J. Biol. Chem. 255:5184–5189.

    PubMed  Google Scholar 

  35. Ilan, Z., Ilan, R., and Cinti, D. L. 1981. Evidence for a new physiological role of hepatic NADPH:ferricytochrome (P-450) oxidoreductase. Direct electron input to the microsomal fatty acid chain elongation system. J. Biol. Chem. 256:10066–10072.

    PubMed  Google Scholar 

  36. Trakshel, G. M., Kutty, R. K., and Maines, M. D. 1988. Resolution of the rat brain heme oxygenase activity: absence of a detectable amount of the inducible form (HO-1). Arch. Biochem. Biophys. 260:732–739.

    PubMed  Google Scholar 

  37. Kapitulnik, J., Gelboin, H. V., Guengerich, F. P., and Jacobowitz, D. M. 1987. Immunohistochemical localization of cytochrome P-450 in rat brain. Neuroscience 20:829–833.

    PubMed  Google Scholar 

  38. Van der Hoeven, T., and Galivan, J. 1987. The effect of dexamethasone, insulin and triiodothyronine on microsomal NADPH-cytochrome-c (P-450) reductase in primary cultures of isolated hepatocytes. Biochem. Biophys. Acta 931:59–67.

    PubMed  Google Scholar 

  39. Ghersi-Egea, J. F., Walther, B., Perrin, R., Minn, A., and Siest, G. 1987. Inducibility of rat brain drug-metabolizing enzymes. Eur. J. Drug Metab. Pharmacokinet. 12:263–265.

    PubMed  Google Scholar 

  40. Ghersi-Egea, J. F., Minn, A., and Siest, G. 1988. A new aspect of the protective function of the blood-brain barrier: activities of four drug-metabolizing enzyme in isolated rat brain microvessels. Life Sci. 42:2515–2523.

    PubMed  Google Scholar 

  41. Hodgson E. 1987. Introduction to toxicology. Pages 1–22,in Hodgson E. and Levi P. E. (eds.) A Textbook of Modern Toxicology, Elsevier, New York.

    Google Scholar 

  42. Reed, D. J. 1985. Cellular defense mechanisms against reactive metabolites. Pages 71–108,in Anders, M. W. (ed.) Bioactivation of Foreign Compounds, Academic Press, London.

    Google Scholar 

  43. Pellegrini-Giampietro D. E., Cherici G., Alesiani M., Carlà V., and Moroni F. 1988. Excitatory amino acids release from rat hippocampal slices as a consequence of free-radical formation. J. Neurochem. 51:1960–1963.

    PubMed  Google Scholar 

  44. Leibowitz, B. E., and Siegel, B. V. 1980. Aspects of free radical reactions in biological systems: aging. J. Gerontol. 35:45–56.

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Centre du Médicament, CNRS URA 597, Université de Nancy-1, 30 rue Lionnois, 54000, Nancy, France

    J. F. Ghersi-Egea, A. Minn, Z. Jayyosi, V. Arnould, H. Souhaili-El Amri & G. Siest

  2. INSERM U 272, 30 rue Lionnois, 54000, Nancy, France

    J. L. Daval

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  1. J. F. Ghersi-Egea
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  2. A. Minn
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  3. J. L. Daval
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Ghersi-Egea, J.F., Minn, A., Daval, J.L. et al. NADPH:cytochrome P-450(c) reductase: Biochemical characterization in rat brain and cultured neurons and evolution of activity during development. Neurochem Res 14, 883–887 (1989). https://doi.org/10.1007/BF00964819

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