title: Toxic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in developing red seabream (Pagrus major) embryo: an association of morphological deformities with AHR1, AHR2 and CYP1A expressions.
abstract: The toxicity of dioxins such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is mainly mediated by the aryl hydrocarbon receptor (AHR), which regulates the multiple target genes including cytochrome P4501A (CYP1A). In general, bony fishes, which possess at least two distinct AHRs are one of the most sensitive vertebrates to TCDD in early life stage. However, the physiological and toxicological roles of piscine multiple AHRs are not fully understood, especially in marine fish. To understand which AHR is responsible for TCDD toxicity in a marine fish species, we characterized the early life stage toxicity related to the expression of AHRs and CYP1A in red seabream (Pagrus major). The embryos at 10h post-fertilization (hpf) were treated with 0-100 microg/L TCDD for 80 min waterborne exposure. TCDD dose-dependently elicited developmental toxicities including mortality, yolk sac edema, retarded body growth, spinal deformity, reduced heart rate, shortened snout, underdeveloped fin, heart, and lower jaw. Intriguingly, hemorrhage and pericardium edema, typical TCDD developmental defects noticed in other fish species, were not found in red seabream until test termination. The EC(egg)50s for yolk sac edema, underdeveloped fin, and spinal deformity were 170, 240, and 340 pg/g, respectively. The LC(egg)50 was 360 pg/g embryo, indicating that this species is one of the most sensitive fishes to TCDD toxicity. The expression levels of rsAHR1, rsAHR2 and CYP1A mRNAs were also determined in different developmental stages. The rsAHR2 mRNA expression dose-dependently increased following TCDD exposure, while rsAHR1 mRNA level was not altered. Level of rsAHR2 mRNA measured by two-step real-time PCR was 30 times higher than rsAHR1 in embryos treated with the highest dose. Temporal patterns of rsAHR2 and CYP1A mRNAs were similar in TCDD-treated embryos, representing a significant positive correlation between rsAHR2 and CYP1A mRNA levels, but not between rsAHR1 and CYP1A. In comparison of temporal trends of TCDD-induced AHRs and CYP1A expression, and developmental toxicities, the highest expression of rsAHR2 and CYP1A mRNA were detected prior to the appearance of maximal incidence of TCDD toxic manifestations. These results suggest that rsAHR2 may be dominantly involved in the transcriptional regulation of CYP1A, and several TCDD defects are dependent on the alteration of rsAHR2 and/or rsAHR2-CYP1A signaling pathway that is controlled through their expression levels.
authors.affiliation: Ehime Prefectural Institute of Public Health and Environmental Science, Matsuyama 790-0003, Japan. yamauchi-masanobu@pref.ehime.jp
authors.forename: Masanobu
authors.initials: M
authors.lastname: Yamauchi
authors.forename: Eun-Young
authors.initials: EY
authors.lastname: Kim
authors.forename: Hisato
authors.initials: H
authors.lastname: Iwata
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Shima
authors.forename: Shinsuke
authors.initials: S
authors.lastname: Tanabe
mesh_terms: D000818:Animals; D019363:Cytochrome P-450 CYP1A1 / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004625:Embryo, Nonmammalian / Q000002:abnormalities / Q000187:drug effects*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D048788:Growth and Development / Q000187:drug effects; D007928:Lethal Dose 50; D000072317:Polychlorinated Dibenzodioxins / Q000032:analysis / Q000633:toxicity*; D018336:Receptors, Aryl Hydrocarbon / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D021541:Sea Bream / Q000002:abnormalities / Q000196:embryology* / Q000502:physiology; D013045:Species Specificity; D016019:Survival Analysis; D013997:Time Factors; D014874:Water Pollutants, Chemical / Q000633:toxicity*; D029961:Zebrafish Proteins / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics; D015053:Zygote / Q000187:drug effects
chemical_list: C474372:AHR2 protein, zebrafish; D017931:DNA Primers; D000072317:Polychlorinated Dibenzodioxins; D018336:Receptors, Aryl Hydrocarbon; D014874:Water Pollutants, Chemical; D029961:Zebrafish Proteins; D019363:Cytochrome P-450 CYP1A1