Abstract
Studies initiated to investigate the presence of cytochrome P4503A (CYP3A) isoenzymes in brain revealed constitutive mRNA and protein expression of CYP3A1 in rat brain. Western blotting studies showed that pretreatment with CYP3A inducer such as pregnenolone-16α -carbonitrile (PCN) significantly increased the cross reactivity comigrating with hepatic CYP3A1 and CYP3A2 in rat brain microsomes. RT-PCR studies have also shown increase in mRNA expression of CYP3A1 following pretreatment of rats with PCN. The ability of rat brain microsomes to catalyze the demethylation of erythromycin, known to be mediated by CYP3A isoenzymes in liver and significant increase in the activity of erythromycin demethylase (EMD) following pretreatment with dexamethasone or PCN have indicated that CYP3A isoenzymes expressed in brain are functionally active. Kinetic studies revealed that increase in the enzyme activity following pretreatment with PCN resulted in increase in the apparent affinity (Km) and Vmax of the reaction. Similarities in the inhibition of the constitutive and inducible brain and liver EMD activity following in vitro addition of ketoconazole, a inhibitor specific for CYP3A catalysed reactions and anti-CYP3A have further indicated that like in liver, CYP3A isoenzymes catalyse the activity of EMD in rat brain. Data also revealed regional differences in the activity of EMD in the brain. Relatively higher constitutive as well as inducible mRNA expression of CYP3A1 in hypothalamus and hippocampus, the brain regions responsive to steroid hormones have suggested that CYP3A isoenzymes may not only be involved in the process of detoxication mechanism but also in the metabolism of endogenous substrates in brain.
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References
Guengerich FP, Shimada T: Oxidation of toxic and carcinogenic chemicals by human cytochrome P450 enzymes. Chem. Res. Toxicol 4: 391–407, 1991
Schutz EG: Induction of Cytochrome P450s. Current Drug Metabolism 2: 139–147, 2001
de Waziers J, Cugnenc PH, Yang CS, Leroux JP, Beaune PH: Cytochrome P450 isoenzymes, epoxide hydrolase and glutathione transferases in rat and human hepatic and extrahepatic tissues. J. Pharmacol. Exp. Ther. 253: 387–394, 1990
Cholerton S, Daly AK, Idle JR: The role of individual human cytochrome P450 in drug metabolism and clinical response. TIPS 13: 434–439, 1992
Evans WE, Relling MV: Pharmacogenomics: translating functional genomics into rational therapeutics. Science 286: 487–491, 1999
Eyles DW, McGrath JJ, Pond SM: Formation of pyridinium species of haloperidol in human liver and brain. Psychopharmacology (Berl) 125: 214–219, 1996
Guengerich FP, Gillam EMJ, Martin MV, Baba T, Kim BR, Shimada T, Raucy KD, Yun CH: The importance of cytochrome P450 3A enzymes in drug metabolism, in: MR Waterman, Hildebrand M (Eds.), Scherring Foundation Workshop 13, Assessment of the use of Single cytochrome P450 enzymes in Drug Research, Springer-Verlag, Berlin,161–186, 1994
Rendic S, Di Carlo FJ: Human cytochrome P450 enzymes: a status report summarizing their reactions, substrates, inducers, and inhibitors. Drug Metab. Rev 29: 413–580, 1997
Sequeira DJ, Strobel HW: In vitro metabolism of imipramine by brain microsomes: effects of inhibitors and exogenous cytochrome P450 reductase. Brain Res 738: 24–31, 1996
Wrighton SA, Schuetz EG, Thummel KE, Shen DD, Korzekwa KR, Watkins PB: The human CYP3A subfamily: practical considerations. Drug Metab Rev 32: 339–361, 2000
Quattrochi L, Guzelian P: CYP3A regulation: from pharmacology to nuclear receptors. Drug Metabolism and Disposition 29: 615–622, 2001
Tang C, Chiba M, Nishime J, Hochman JH, Chen I, Williams TM, Lin JH: Comparison of imidazole- and 2-methyl imidazole-containing farnesyl-protein transferase inhibitors: interaction with and metabolism by rat hepatic cytochrome P450s. Drug Metab Dispos 28: 680–686, 2000
Burk O, Wojnowski L.: Cytochrome P4503A and their regulation, Naunyn-Schmiedeberg's Arch Pharmacol 369:105–124, 2004
Mcfadyen MCE, Melvin WT, Murray GI: Regional distribution of individual forms of cytochrome P450 mRNA in normal adult human brain. Biochem. Pharmacol 55: 825–830, 1998
Rosenbrock H, Hagemeyer CE, Ditter M, Knoth R, Volk B: Identification, induction and localization of cytochrome P450s of the 3A-subfamily in mouse brain. Neurotox Res 3: 339–349, 2001
Schilter B, Omiecinski CJ: Regional distribution and expression modulation of cytochrome P-450 and epoxide hydrolase mRNAs in the rat brain. Mol Pharmacol 44: 990–6, 1993
Jayyosi Z, Cooper KO, Thomas PE: Brain cytochrome P450 and testosterone metabolism by rat brain subcellular fractions: presence of cytochrome P450 3A immunoreactive protein in rat brain mitochondria. Arch Biochem Biophys 298: 265–70, 1992
Wang H, Kawashima H, Strobel HW: cDNA Cloning of a Novel CYP3A from Rat Brain. Biochem and Biophys Res Comm 221: 157–162, 1996
Hagemeyer CE, Rosenbrock H, Ditter M, Knoth R, Volk B: Influence of exogenous compounds on endogenous metabolic pathways by members of the CYP3A subfamily in mouse brain. Acta Neuropathol 102: 525–526, 2001
Hagemeyer CE, Rosenbrock H, Ditter M, Knoth R, Volk B: Predominantly neuronal expression of cytochrome P450 isoforms CYP3A11 and CYP3A13 in mouse brain. Neuroscience 117: 521, 2003
Igarashi K, Kasuyal F, Fukuil M, Usuki E, Castagnoli Jr N: Studies on the metabolism of haloperidol (HP): the role of CYP3A in the production of the neurotoxic pyridinium metabolite HPP+ found in rat brain following i.p administration of HP. Life Sci 57: 2439–2446, 1995
Omiecinski, C. J. Hassett, C. Costa, P: Developmental expression and in situ localization of the phenobarbital-inducible rat hepatic mRNAs for cytochromes CYP2B1, CYP2B2, CYP2C6, and CYP3A1. Mol Pharmacol 38: 462–470, 1990
Kawai M, Bandiera SM, Chang TKH and Bellward GD: Growth hormone regulation and developmental expression of rat hepatic CYP3A18, CYP3A9 and CYP3A2. Biochem. Pharmacol 59: 1277–87, 2000
Glowinski J, Iversen LL: Regional studies of catecholamines in the rat brain. I. The disposition of [3H] norepinephrine, [3H] dopamine and [3H] dopa in various regions of the brain. J Neurochem 13: 655–669, 1966
Parmar D, Dhawan A and Seth PK: Immunochemical evidence for the presence of Phenobarbital (PB) and 3-methylcholanthrene (MC) inducible cytochrome P450 isoenzymes in rat brain. Int. J. Toxicol 17: 619–630, 1998
Chomczynski P, Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159, 1987
Arlotto MP, Sonderfan AJ, Klassen CD, Parkinson A: Studies on the pregnenolone-16-a-caronitrile-inducible form of rat liver microsomal cytochrome P450 and UDP-glucuronosyltransferase. Biochem. Pharmacol 36: 3859–3866, 1987
Soh Y, Rhee HM, Sohn DH, Song BJ: Immunological detection of CYP2E1 in fresh rat lymphocytes and Its pretranslational induction by fasting. Biochem and Biophys Res Comm 227: 541–546, 1996
Hagemeyer CE, Rosenbrock H, Singec I, Knoth R, Volk B: Different testosterone metabolism by immortalized embryonic and postnatal hippocampal neurons from C57BL/6 mice: a crucial role for androstenedione. J Neurosci Res 60: 106–15, 2000
Thuerlm C, Otten U, Knoth R, Meyer RP, Volk B: Possible role of cytochrome P450 in inactivation of testosterone in immortalized hippocampal neurons. Brain Res 762: 47–55, 1997
Rosenbrock H, Hagemeyer CE, Singec I, Knoth R, Volk B: Testosterone metabolism in rat brain is differentially enhanced by phenytoin-inducible cytochrome P450 isoforms. J Neuroendocrinol 11: 597–604, 1999
Pai HV, Upadhya SC, Chinta SJ, Hegde SN, Ravindranath V: Differential metabolism of alprazolam by liver and brain cytochrome (P4503A) to pharmacologically active metabolite. Pharmacogenomics J 2: 243–258, 2002
Souidi M, Gueguen Y, Linard C, Dudoignon N, Grison S, Baudelin C, Marquette C, Gourmelon P, Aigueperse J, Dublineau I: In vivo effects of chronic contamination with depleted uranium on CYP3A and associated nuclear receptors PXR and CAR in the rat. Toxicology. 2005 (Article in press)
Lamba V, Yasuda K, Lamba JK, Assem M, Davila J, Strom S, Schuetz EG: PXR (NR1I2): splice variants in human tissues, including brain, and identification of neurosteroids and nicotine as PXR activators. Toxicol Appl Pharmacol 199: 251–265, 2004
Mei Q, Richards K, Strong-Basalyga K, Fauty SE, Taylor A, Yamazaki M, Prueksaritanont T, Lin JH, Hochman J: Using real-time quantitative TaqMan RT-PCR to evaluate the role of dexamethasone in gene regulation of rat P-glycoproteins mdr1a/1b and cytochrome P450 3A1/2.J Pharm Sci 93: 2488–2496, 2004
Martinez C, Gervasini G, Agundez JA, Carrillo JA, Ramos SI, Garcia-Gamito FJ, Gallardo L, Benitez J: Modulation of midazolam 1-hydroxylation activity in vitro by neurotransmitters and precursors. Eur J Clin Pharmacol 56: 145–151, 2000
LaBella FS, Brandes LJ: Interaction of histamine and other bioamines with cytochrome P450: implications for cell growth modulation and chemopotentiation by drugs. Seminars in Cancer Biology 10: 47–53, 2000
Tamasi V, Fulop AK, Hegyi K, Monostory K, Falus A: Upregulation of CYP2e1 and CYP3a activities in histamine-deficient histidine decarboxylase gene targeted mice. Cell Biol Int 27: 1011–1015, 2003
Robertson GR, Field J, Goodwin B, Bierach S, Tran M, Lehnert A, Liddle C: Transgenic mouse models of human CYP3A4 gene regulation. Mol Pharmacol 64: 42–50, 2003
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Yadav, S., Dhawan, A., Seth, P.K. et al. Cytochrome P4503A: Evidence for mRNA expression and catalytic activity in rat brain. Mol Cell Biochem 287, 91–99 (2006). https://doi.org/10.1007/s11010-005-9080-8
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DOI: https://doi.org/10.1007/s11010-005-9080-8