Exposure to Endocrine-Disrupting Chemicals (EDCs) is suspected to be the cause for the increasing number of reproductive disorders in wildlife. It is well established that many chemicals in the environment can interfere with signalling mechanisms/hormonal regulation of fish at concentrations considerably below those causing cellular toxicity. Key targets for EDCs include steroid hormone receptors and steroidogenic enzyme systems. Cytochrome P450arom (CYP19A1 gene), a key enzyme in the hormonal steroidogenic pathway, mediates the conversion of androgens to estrogens. This work aimed to study the effects of EDCs on the expression of CYP19A1 mRNA in the common carp (Cyprinus carpio). For this purpose, we have cloned a full-length cDNA encoding cytochrome P450 aromatase. In particular we have used degenerated primers designed on the conserved regions of CYP19A1 of other teleost species. On the basis of such sequence we have designed PCR primers for the semi-quantitative PCR assays. RT-PCR revealed that the CYP19A1 was highly expressed in the ovary and, at very low levels, in the testis. Effects of a 14 day exposure (OECD guideline 204) to graded concentrations of bisphenol A (BPA) on the expression of mRNA CYP19A1 are discussed
Cloning of mRNA CYP19A1 in the carp (Cyprinus carpio)
MANDICH, ALBERTA;MASSARI, ALESSANDRA;CANDIANI, SIMONA;CEVASCO, ALESSANDRA;BOTTERO, SERGIO;PEDEMONTE, FEDERICO
2006-01-01
Abstract
Exposure to Endocrine-Disrupting Chemicals (EDCs) is suspected to be the cause for the increasing number of reproductive disorders in wildlife. It is well established that many chemicals in the environment can interfere with signalling mechanisms/hormonal regulation of fish at concentrations considerably below those causing cellular toxicity. Key targets for EDCs include steroid hormone receptors and steroidogenic enzyme systems. Cytochrome P450arom (CYP19A1 gene), a key enzyme in the hormonal steroidogenic pathway, mediates the conversion of androgens to estrogens. This work aimed to study the effects of EDCs on the expression of CYP19A1 mRNA in the common carp (Cyprinus carpio). For this purpose, we have cloned a full-length cDNA encoding cytochrome P450 aromatase. In particular we have used degenerated primers designed on the conserved regions of CYP19A1 of other teleost species. On the basis of such sequence we have designed PCR primers for the semi-quantitative PCR assays. RT-PCR revealed that the CYP19A1 was highly expressed in the ovary and, at very low levels, in the testis. Effects of a 14 day exposure (OECD guideline 204) to graded concentrations of bisphenol A (BPA) on the expression of mRNA CYP19A1 are discussed
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