title: Effects of the polycyclic ketone tonalide (AHTN) on some cell viability parameters and transcription of P450 and immunoregulatory genes in rainbow trout RTG-2 cells.
abstract: To assess the potential effects of environmental pollutants belonging to the musk fragrances group in the physiology of aquatic animal species, in this work we treated rainbow trout RTG-2 cells with the polycyclic ketone tonalide (AHTN) at dilutions ranging from 3.5 to 500 ng/ml. The following parameters were monitored: intracellular ATP concentration (energy production), mitochondrial membrane potential (early apoptosis marker), cell viability (vital staining with DFP), quantitative expression of genes coding for the cytochrome P450 detoxifying enzymes CYP1A1 and CYP3A27, and of genes coding for the immunoregulatory peptides IL-1β, IL-8, TNFα, Cox-2 and TGF-β. Obtained results showed that incubation with tonalide induced in RTG-2 cells no effects on cell viability, a slight increase of mitochondrial membrane potential activity, and a significant increase in intracellular ATP concentration. However, dramatic effects were observed in transcription levels of some tested genes, with upregulation levels of 300 and 600 times measured for TGF-β and TNFα, respectively and of 150 times for the CYP3A27 gene. Our results show for the first time the potent effects exerted by tonalide on immunoregulatory genes of RTG-2 cells and also indicate that the measured sensitivity of RTG-2 towards tonalide was in the same range of that currently available using chemical methods. A possible use of the panel of genes we employed as a tool for the monitoring of musk fragrances in biological samples is discussed.
authors.affiliation: Department for Innovation in Biological, Agro-Food and Forest Systems, Università della Tuscia, Viterbo, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Randelli
authors.forename: Valerio
authors.initials: V
authors.lastname: Rossini
authors.forename: Ilaria
authors.initials: I
authors.lastname: Corsi
authors.forename: Silvano
authors.initials: S
authors.lastname: Focardi
authors.forename: Anna M
authors.initials: AM
authors.lastname: Fausto
authors.forename: Francesco
authors.initials: F
authors.lastname: Buonocore
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Scapigliati
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics; D002232:Carbocyanines / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D051546:Cyclooxygenase 2 / Q000235:genetics; D003603:Cytotoxins / Q000633:toxicity*; D005452:Fluoresceins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D053583:Interleukin-1beta / Q000235:genetics; D016209:Interleukin-8 / Q000235:genetics; D017686:Oncorhynchus mykiss / Q000235:genetics*; D010476:Perfume / Q000633:toxicity*; D012313:RNA / Q000096:biosynthesis; D013764:Tetrahydronaphthalenes / Q000633:toxicity*; D014158:Transcription, Genetic / Q000187:drug effects*; D016212:Transforming Growth Factor beta / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics; D014874:Water Pollutants, Chemical / Q000633:toxicity*
chemical_list: D002232:Carbocyanines; D003603:Cytotoxins; D005452:Fluoresceins; D005456:Fluorescent Dyes; D053583:Interleukin-1beta; D016209:Interleukin-8; D010476:Perfume; D013764:Tetrahydronaphthalenes; D016212:Transforming Growth Factor beta; D014409:Tumor Necrosis Factor-alpha; D014874:Water Pollutants, Chemical; C033116:acetyl methyl tetramethyl tetralin; C007392:3,3'-dihexyl-2,2'-oxacarbocyanine; D012313:RNA; D000255:Adenosine Triphosphate; D001189:Aryl Hydrocarbon Hydroxylases; D051546:Cyclooxygenase 2; C018506:diacetylfluorescein