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ToxSci Advance Access published online on July 27, 2006
Toxicological Sciences, doi:10.1093/toxsci/kfl072
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© The Author 2006. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
Received July 21, 2006
Accepted July 24, 2006

Environmental Toxicology

CYP1C1 mRNA Expression is Inducible by Benzo(a)pyrene in Fundulus heteroclitus Embryos and Adults

Lu Wang 1, Brian E. Scheffler 2, and Kristine L. Willett 1 *

1 Department of Pharmacology and Environmental Toxicology Research Program, School of Pharmacy, University of Mississippi, University, MS 38677
2 USDA-ARS-CGRU MSA Genomics Laboratory, 141 Experiment Station Rd., Stoneville, MS 38776

* To whom correspondence should be addressed.
Kristine L. Willett, E-mail: kwillett{at}olemiss.edu


  Abstract

CYP1C is the newest member of the CYP1 family of P450s, however its physiological significance, its inducers and metabolic functions are unknown. Two full-length alleles of Fundulus heteroclitus CYP1C1 cDNA were cloned. The 529 amino acid protein shared the highest amino acid identity with Stenotomus chrysops CYP1C1 (81%). To investigate whether the carcinogen benzo(a)pyrene (BaP) was a CYP1C1 inducer, we used real time PCR to quantitatively measure tissue and sex specific expression of both CYP1C1 and CYP1A mRNAs in BaP exposed adult fish. CYP1C1 mRNA expression was constitutively higher than CYP1A in brain, spleen, eye and gonad, while CYP1A was higher in gastrointestinal tract (GI), heart, gill and liver. Kidney had equal but high expression of both CYP1s. There were sex differences in constitutive CYP1 expression in the GI, liver, gill, and eye. BaP exposure caused induction of CYP1C1 expression in female and male heart (31 and 17-fold), gill (7 and 4-fold) and liver (6 and 5-fold), respectively. Embryo CYP1 expression was constitutively highest at two weeks post hatch, and whole embryos expressed 3 to 15-fold more CYP1C1 mRNA compared to CYP1A. BaP, 10 µg/L for 10 days, caused induction of both genes at 120 and 240 hpf. Our results suggest that teleost CYP1C, in addition to CYP1A, is inducible by BaP, has a broad tissue distribution, and should be further investigated for its role in carcinogen bioactivation.

Keywords: CYP1C1; Cytochrome P450; Benzo(a)pyrene; Killifish; Embryos; Fundulus.
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