Abstract
Metabolism of therapeutic drugs in the body by the mixed function oxidase system is an important consideration in the analysis of a drug's effectiveness. P450-dependent metabolism within the brain of a neuro-specific drug may affect the drug's course of action. To determine whether cytochrome P450 was expressed in brain, RNA was isolated from the whole brains of rats treated with a variety of known hepatic P450 inducers, including amitriptyline, imipramine, isosafrole, phenobarbital, and β-naphthoflavone. The RNA was analyzed for the presence of P450 isozymes by the PCR technique. Differential expression of P450IA1, P450IIB1, P450IIB2, P450IID, and P450IIE1 was detected in the brain samples, depending on the treatment. Cytochrome P450 reductase expression was also detected in the brain samples, giving strong evidence that the brain contains a competent mixed function oxidase system under all conditions studied. (Mol Cell Biochem120: 171–179, 1993)
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Strobel HW, Lu AYH, Heidemak J, Coon MJ: Phosphatidylcholine requirement in the enzymatic reduction of hemeprotein P450 and in fatty acid, hydrocarbon and drug hydroxylation. J Biol Chem 245: 4851–4854, 1970
Gillette JR: Biochemistry of drug oxidation and reduction by enzymes in hepatic endoplasmic reticulum. Adv Pharmacol 4: 219–216, 1966
Conney AH: Pharmicological implications of microsomal enzyme induction. Pharmacol Rev 19:317–366, 1967
Nebert DW: Genes encoding drug-metabolizing enzymes: Possible role in human disease. In: AD Woodhead, MA Bender, RC Leonard (eds.) Phenotypic Variation in Populations, Plenum Publishing Corperation, New York, 1988, pp 45–64
Fang WF, Strobel HW: The drug and carcinogen metabolism system of rat colon microsomes. Arch Biochem Biophys 186: 128–138, 1978
Guengerich FP: Preparation and properties of highly purified cytochrome P450 and NADPH-cytochrome P450 reductase from pulmonary microsomes of untreated rabbits. Mol Pharmacol 13: 911–923, 1977
Cohn JA, Alvares AP, Kappas A: On the occurrence of cytochrome P450 and aryl hydrocarbon hydroxylase activity in rat brain. J Exp Med 145: 1607–1611, 1977
Sasame HA, Ames MM, Nelson SD: Cytochrome P450 and NADPH cytochrome c reductase in rat brain: formation of catechols and reactive catechol metabolites. Bio Biol Res Comm 78: 919–929, 1977
Levitt RC, Fysh JM, Jenson NM, Nebert DW: The {iAH} locus: Biochemical basis for genetic differences in brain tumor formation in mice. Genetics 92: 1205–1210, 1979
Barbeau A, Cloutier T, Roy M, Plasse L, Paris S, Poirier J: Ecogenetics of Parkinson's Disease: 4-hydroxylation of debrisoquine. Lancet 2, 1213–1216, 1985
Shahi GS, Das NP, Moochhals SM: Parkinson's disease and cytochrome P450: a possible link? Med Hypotheses 32: 277–282, 1990
Fonne-Pfister R, Bargetzi MJ, Meyer UA: MPTP, the neurotoxin inducing Parkinson's disease, is a potent competitive inhibitor of human and rat cytochrome P450 isozymes (P450bufI, P450db1 (P450IID6)) catalyzing debrisoquine 4-hydroxylation. Biochem Biophys Res Commun 148: 1144–1150, 1987
Naslund BM, Glaumann H, Warner M, Gustafsson JA, Hansson T: Cytochrome P450 b and c in the rat brain and pituitary gland. Mol Pharmacol 33: 31–37, 1988
Tyndale F, Sunahare R, Inaba T, Kalow W, Gonzalez FJ, Niznik HB: Neuronal cytochrome P450IID1 (debrisaquine/sparteinetype): potent nucleotide sequence identity to human hepatic P450 gene cyp2D6. Mol Pharmacol 40: 63–68, 1991
Ravindranath V, Anandatheerthavarada HK, Shankar SK: Xenobiotic metabolism in human brain-presence of cytochrome P450 and associated mono-oxygenases. Brain Res 496: 331–335, 1989
Kapitulnik J, Gelboin HV, Guengerich FP, Jacobowitz DM: Immunohistochemical localization of cytochrome P450 in rat brain. Neuroscience 20: 829–833, 1987
Kohler C, Erikson LG, Hansson T, Warner M, Ake-Gustafsson J: Immunohistochemical localization of cytochrome P-450 in the rat brain. Neurosci Lett 84: 109–144, 1988
Warner M, Kohler C, Hannson T, Gustafsson JA: Regional distribution of cytochrome P-450 in the rat brain: spectral quantitation and contribution of P-450b, e and P-450c, d. J Neurochem 50: 1057–1065, 1988
Hansson T, Tinberg N, Ingelman-Sundberg M, Kohler C: Regional distribution of ethanol-inducable cytochrome P-450IIE1 in the rat central nervous system. Neuroscience 34: 451–463, 1990
Haglund L, Kohler C, Haaparanta T, Goldstein M, Gustafsson JA: Immunohistochemical evidence for a heterogeneous distribution of NADPH cytochrome P450 reductase in the rat and monkey brain. In: J Rydstrom, J Montelius, M Bengtsson (eds.) Extrahepatic Drug Metabolism and Chemical Carcinogensis. Elsebier Science Publishers B.V., New York, 1983, pp 89–93
Takeshita M, Miki M, Yubisui T: Cytochrome P450 reductase activities in rat brain microsomes during development. J Neurochem 39: 1047–1049, 1982
Bergh A, Strobel HW: Reconstitution of the brain mixed function oxidase system: purification of NADPH cytochrome P-450 reductase and partial purification of cytochrome P450 from whole brain. J Neurochem 59: 576–581, 1992
Li H, Gyllensten UB, Cui X, Saiki RK, Erlich HA, Arnheim N: Amplification and Analysis of DNA Sequences in Single Human Sperm and Diploid Cells. Nature 335: 414–41, 1988
Warner M, Ahlgrin R, Zaphiropoulos PG, Hayashi S, Gustafsson JA: Identification and Localization of Cytochromes P450 Expressed in Brain. In: MR Waterman, EF Johnson (eds.) Methods of Enzymology, vol. 206. Academic Press, New York, 1991, pp 631–641
Krug MS, Berger SL: First-Strand cDNA Synthesis Primed with Oligo(dT). In: SL Berger, AR Kimmel (eds.) Methods of Enzymology, vol. 152. Academic Press, New York, 1987, pp 316–325
Devereux J, Haeberli P, Smithies O: A Comprehensive Set of Sequences Analysis Programs for the VAX. Nucleic Acids Research 12(1): 387–395, 1984
Author information
Authors and Affiliations
Additional information
Thesis student of the Graduate School of Biomedical Sciences, the University of Texas Health Science Center at Houston
Rights and permissions
About this article
Cite this article
Hodgson, A.V., White, T.B., White, J.W. et al. Expression analysis of the mixed function oxidase system in rat brain by the polymerase chain reaction. Mol Cell Biochem 120, 171–179 (1993). https://doi.org/10.1007/BF00926090
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00926090