CHARACTERIZATION OF RTG-2 FISH CELL-LINE BY RANDOM AMPLIFIED POLYMORPHIC DNA

The increasing presence of genotoxic chemicals in the aquatic environm ent has led to the development of both in vivo and in vitro assays for target species. The fish population represents an important level of aquatic ecosystems that can be threatened by increased environmental p ollution. The authors have studied the DNA pattern of the RTG-2 fish c ell line, a fibroblast-like cell line, derived from rainbow trout (Onc orhynchus mikyss), to use this cell line as an in vitro system to stud y genotoxicity by means of random amplified polymorphic DNA primers (R APDs). A constant pattern in the DNA band is essential when an organis m or cell line is used to detect DNA alterations produced by genotoxic environmental chemicals. DNA fingerprints with RAPDs were obtained fo r RTG-2 by testing 26 single and 70 pairwise combinations of primers. Different methods of DNA extraction (chelating resin, salting out, and phenolization), the influence of spectrometric measures at 320 nm in the 260/280 quotient to quantify DNA extracts, genomic DNA and primer concentrations, annealing temperatures, and cell line passage were stu died in the cell line characterization. RAPD products were identified by agarose gel electrophoresis. The good results obtained should allow the use of this system as a possible tool for detection of the genoto xicity of aquatic pollutants. (C) 1998 Academic Press.