Differential gene expression in anthracene-exposed mummichogs (Fundulus heteroclitus).
The polycyclic aromatic hydrocarbon (PAH) anthracene is present in many estuarine systems at concentrations believed to cause sublethal adverse effects, although its exact mode of toxicity remains unclear. Knowledge of the induction or suppression of specific genes as a result of exposure may be useful in explaining these effects. We have generated a fingerprint of anthracene exposure using the mummichog (Fundulus heteroclitus), a non-migratory estuarine fish species. The fish were exposed in 7-day static renewal tests to environmentally relevant concentrations of 0, 27, 50, and 80 microg/l of anthracene. Total RNA was extracted from the livers and differential display reverse transcription polymerase chain reaction (DD RT-PCR) was used to recover 26 differentially expressed cDNA fragments. These cDNAs were isolated, sequenced, and compared to sequences of known genes in order to identify possible physiological consequences of exposure to anthracene. We then constructed macroarrays using these fragments and probed them with RNA from both anthracene-exposed fish and fish from a known PAH-impacted site. Three genes appear to be good indicators of exposure to anthracene in the range of concentrations tested, which included CYP2N2 and two expressed sequence tags (ESTs) termed 15C1 and 18C2. The expression of nine genes was altered in fish collected from a site with multiple PAHs. Band 15C1 and CYP2N2 again showed statistically significant upregulation in the field-caught fish, while a trypsin precursor and fatty acid-binding protein (FABP) all showed similar trends in induction as the laboratory-exposed fish. Further insight into the mechanism of toxicity of contaminants will be gained by the ability to identify and use differentially expressed genes as markers of exposure and effects.
Peterson
Janis S K
JS
Department of Biological Sciences, University of Texas at El Paso, 500 W University Avenue, El Paso, TX 79968, USA.
Bain
Lisa J
LJ
D000704:Analysis of Variance; D000818:Animals; D000873:Anthracenes / Q000032:analysis / Q000633:toxicity*; D001483:Base Sequence; D002352:Carrier Proteins / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D000072467:Cytochrome P450 Family 2; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D020224:Expressed Sequence Tags; D050556:Fatty Acid-Binding Proteins; D023781:Fundulidae / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects*; D008969:Molecular Sequence Data; D012313:RNA / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D018675:Toxicity Tests
D000873:Anthracenes; D002352:Carrier Proteins; D017931:DNA Primers; D018076:DNA, Complementary; D050556:Fatty Acid-Binding Proteins; D012313:RNA; D003577:Cytochrome P-450 Enzyme System; D000072467:Cytochrome P450 Family 2; C404183:cytochrome P-450 CYP2N2 (killifish); C034020:anthracene