DNA Adducts as Early Bioindicators of Chemical Exposure

Abstract

Sensitive analytical methods indicate the presence of hundreds of chemical contaminants in our environment. However, concentration of these pollutants is usually at the low parts per million or parts per billion level. At such low levels, toxicants induce long-term (chronic) rather than short-term (acute) toxicities. Experiments are designed to evaluate chronic toxicity using early bioindicators. Recently, fish have been used as experimental animals because some species show early (weeks as opposed to years in other research animals), sensitive responses. Thus, medaka (Oryzias latipes), following exposure to diethylnitrosamine (DEN), exhibited liver tumors in several weeks; more interestingly its DNA was modified (to a ‘DNA-adduct’) after only 24-48 hours exposure. Such adducts show promise as early bioindicators because they are formed within hours of exposure. The formation of DNA-adducts were monitored in medaka and rainbow trout (Salmo gairdneri) exposed to DEN by aqueous or intra-peritoneal routes. Using HPLC-Fluorimetry, O ₆-ethylguanine (0 ₆-EtGua) was detected and monitored in acid thermal hydrolysates of DNA isolates. (Detection limit for O ₆-EtGua was as low as 3 ng, 1.7 X 10_-11 moles). Fourier transform cyclotron resonance mass spectrometry with IR laser desorption/ionization was used advantageously to establish the structure of nucleotides, bases and nucleosides directly, without further cumbersome derivatizations. Other aspects of DNA-adducts are discussed.

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