No detectable DNA excision repair in UV-exposed hepatocytes from two catfish species.
DNA repair is a critical process in protecting cellular genetic information from mutation. Nucleotide excision repair (NER) is a mechanism by which cells correct DNA damage caused by agents that form bulky covalent adducts and UV photoproducts such as thymine dimers and 6-4 photoproduct. NER, sometimes called dark repair, is generally accepted as being low in fish compared to mammals. This study was designed to quantitate NER in two related catfish species that have known differential sensitivities to liver carcinomas. The original hypothesis was that the more cancer resistant species, channel catfish (Ictalurus punctatus), would have more efficient DNA repair compared to the more sensitive brown bullhead (Ameriurus nebulosus). In order to measure NER, primary cultured hepatocytes of both species were exposed to UV light (10-40 J/m2) and collected at 0, 24, 48 and 72 h after exposure. Total DNA was extracted from the cells and incubated with T4 endonuclease V. Using alkaline gel electrophoresis, endonuclease sensitive sites (ESS) were quantified. Results from the ESS assay indicated there was a UV dose-response increase in thymine dimers from 0 to 40 J/m2. However, no repair (decrease in number of ESS) occurred in either fish species over a 72-h time period. When cells were exposed to photoreactivating fluorescent light, repair was detected. These studies highlight the difficulty of measuring NER in fish and are consistent with the low levels of NER reported by other researchers in fish.
Ecotoxicology Laboratory, Nicholas School of the Environment, Duke University, Durham, NC 27708-0328, USA. kwillet@olemiss.edu
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Willett
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Lienesch
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Di Giulio
D000818:Animals; D018929:Cell Culture Techniques; D004260:DNA Repair; D004255:Deoxyribodipyrimidine Photo-Lyase / Q000494:pharmacology; D043224:Deoxyribonuclease (Pyrimidine Dimer); D004307:Dose-Response Relationship, Radiation; D004706:Endodeoxyribonucleases / Q000494:pharmacology; D022781:Hepatocytes / Q000528:radiation effects*; D007059:Ictaluridae / Q000235:genetics*; D008297:Male; D011740:Pyrimidine Dimers / Q000528:radiation effects; D013997:Time Factors; D014466:Ultraviolet Rays / Q000009:adverse effects*; D014764:Viral Proteins
D011740:Pyrimidine Dimers; D014764:Viral Proteins; D004706:Endodeoxyribonucleases; C470698:endonuclease V, phage T4; D043224:Deoxyribonuclease (Pyrimidine Dimer); D004255:Deoxyribodipyrimidine Photo-Lyase