Octylphenol (OP) is the degradative product of alkylphenol ethoxylates that are widely used to produce rubber, pesticides, and paints. It is chemically stable substance and demonstrates estrogenic effects, toxicity and carcinogenic effects in the environment. The toxin accumulates rapidly in the liver where it exerts most of its damage, but the molecular mechanisms behind its toxicity remain unclear. Due to limited information concerning the effect of OP on liver, this study investigates how OP causes hepatotoxicity in liver. Here, suppression subtractive hybridization was used to identify the alterations in gene transcription of the frog (Rana chensinensis) after exposure to OP. After hybridization and cloning, the subtractive cDNA libraries were obtained. At random, 207 positive clones were selected and sequenced from the subtractive libraries, which gave a total of 75 gene fragment sequences. The screening identified numerous genes involved in apoptosis, signal transduction, cytoskeletal remodeling, innate immunity, material and energy metabolism, translation and transcription which were extensively discussed. Two sequenced genes were analyzed further using real time quantitative PCR. The two genes from the library were found to be transcriptionally up-regulated. These results confirmed the successful construction of the subtractive cDNA library that was enriched for the genes that were differentially transcribed in the amphibian liver challenged with OP, and for the first time present the basic data on toxicity effect of OP on liver.
a total of 64 gastropod specimens of 15 species were collected from the coastal waters of Okinawa Prefecture, Japan, and examined for toxicity by means of mouse bioassay. Among the specimens tested, 5 species, Nassarius glans, Nassarius coronatus, Oliva annulata, Oliva concavospira and Zeuxis sp., were toxic. The toxicity scores of N. glans were very high; 39.6-461 MU/g in muscle, and 98.6-189 MU/g in viscera including digestive gland, followed by Zeuxis sp. 12.7 MU/g in whole body , N. coronatus 5.64-11.1 MU/g in whole body , O. annulata 10.8 MU/g in the whole body , O. concavospira 6.65 MU/g in the muscle . Liquid chromatography/mass spectrometry LC-MS revealed that the major toxic component was tetrodotoxin TTX , which accounting for 13-82 of the total toxicity. As for the remaining toxicity in the case of N. glans, 4,9-anhydroTTX, 4-epiTTX and 11-oxoTTX were contributors. Moreover, Niotha albescens showed no toxicity less than 10 MU/g in mouse bioassay, but TTX 5.08 MU/g was detected by LC-MS. Paralytic shellfish poison was not detected in any of the specimens by high-performance liquid chromatography with fluorometric detection HPLC-FLD .
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