A Comparison of Cytochrome P4501A (CYP1A) mRNA Inducibility in Four Species of Atlantic Coast Anadromous Fishes

Abstract

Quantification of levels of cytochrome P4501A (CYP1A) gene expression in sentinel species of fishes has been proposed as a management tool to evaluate contamination of aquatic systems. Based on preliminary studies, we hypothesized that differences in CYP1A mRNA inducibility among individuals, populations, or species might lead to spurious conclusions when using this approach in environmental monitoring programs. To address this possibility, we quantitated and compared CYP1A mRNA induction levels in four species of common Atlantic Coast estuarine fish: smooth flounder, hogchoker, striped bass, and Atlantic tomcod, which were treated with model chemicals (beta naphthoflavone (β-NF), or benzo[a]pyrene at 10 ppm) known to induce CYP1A mRNA, or were exposed to contaminated environments. Species-specific CYP1A DNA probes were generated from PCR (polymerase chain reaction) amplification of genomic DNA using conserved oligonucleotide primers, and, along with cloned rainbow trout and Atlantic tomcod CYP1A cDNA probes were used to quantify CYP1A mRNA levels in northern blot analyses. Successful PCR amplification of CYP1A hybridizable DNA fragments was observed for all four species. Results from northern blot analyses showed large differences in CYP1A mRNA induction among species; only Atlantic tomcod exhibited significant induction of CYP1A mRNA for both chemically treated (97-fold) and environmentally exposed fish (34-fold). Significant, although lower, levels of induction were observed in β-NF treated (14-fold) smooth flounder, but not in environmentally exposed smooth flounder. Only low levels (not significant) of CYP1A gene induction were detected in hogchokers and striped bass. We conclude that CYP1A mRNA inducibility differed significantly among fish taxa perhaps due to differences in regulation of gene expression, suggesting that careful selection of sentinel species should be exercised prior to the use of CYP1A mRNA induction in environmental monitoring programs. However, the significance of differences in CYP1A mRNA inducibility in relation to higher level biological endpoints has yet to be determined.