title: Persistent sex-reversal and oviducal agenesis in adult Xenopus (Silurana) tropicalis frogs following larval exposure to the environmental pollutant ethynylestradiol.
abstract: It is known that estrogen-like environmental pollutants can feminise gonadal differentiation in frogs resulting in female-biased sex-ratios at metamorphosis. The long-term effects on reproductive function in frogs following larval exposure to pollutants are less known. Amphibian test systems which allow life-cycle studies are therefore needed. The aim of the present study was to characterise long-term estrogenic effects on the reproductive system of the emerging model species Xenopus (Silurana) tropicalis following larval exposure to ethynylestradiol (EE(2)). EE(2) is a synthetic estrogen that has been detected in sewage effluents and in surface waters. Newly hatched tadpoles (Niewkoop Faber (NF) stage 48) were exposed to the nominal EE(2) concentrations 0 (control), 1, 10, and 100 nM (with analytical chemistry support) until complete metamorphosis (NF stage 66). Effects on the reproductive organs were determined in juveniles (1 month after metamorphosis) and in 9-month-old frogs. Larval exposure to EE(2) caused female-biased phenotypic sex-ratios in both juvenile and adult frogs, which is in agreement with previous work on other frog species. Nearly all (97%) of the 63 EE(2)-exposed 9-month-old frogs had ovaries. Histological evaluation of the gonads of the 9-month-old frogs showed that they were sexually mature. Among the adult frogs with ovaries there was a dose-dependent increase in the frequency of individuals lacking oviducts. Adult frogs exposed to 100 nM EE(2) that had ovaries but no oviducts had lower levels of estrogen receptor alpha (ERalpha) mRNA in the brain than control animals and those exposed to 100 nM EE(2) that had ovaries as well as oviducts. EE(2) exposure did not cause any significant changes in ERalpha mRNA levels in the ovaries of the adult frogs. The reduced level of ERalpha mRNA in the brain of individuals with ovaries lacking oviducts suggests an organizing effect of EE(2) on the central nervous system. The results show that transient early life-stage exposure to an environmental pollutant can induce effects on the reproductive organs and the central nervous system that persist into adulthood. Overall, our data suggest that X. tropicalis, which has a shorter generation time than the well-established model species Xenopus laevis, is a suitable model organism for research on developmental reproductive toxicity in anuran species.
authors.affiliation: Department of Environmental Toxicology, Uppsala University, Centre for Reproductive Biology in Uppsala, CRU, Norbyvägen 18 A, 752 36 Uppsala, Sweden.
authors.forename: Irina
authors.initials: I
authors.lastname: Pettersson
authors.forename: Augustine
authors.initials: A
authors.lastname: Arukwe
authors.forename: Katrin
authors.initials: K
authors.lastname: Lundstedt-Enkel
authors.forename: Anne S
authors.initials: AS
authors.lastname: Mortensen
authors.forename: Cecilia
authors.initials: C
authors.lastname: Berg
mesh_terms: D000818:Animals; D001923:Brain Chemistry / Q000187:drug effects; D004781:Environmental Exposure; D047628:Estrogen Receptor alpha / Q000096:biosynthesis / Q000187:drug effects; D004967:Estrogens / Q000633:toxicity; D004997:Ethinyl Estradiol / Q000633:toxicity*; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D007814:Larva / Q000187:drug effects; D008297:Male; D008675:Metamorphosis, Biological / Q000187:drug effects; D010053:Ovary / Q000737:chemistry; D010057:Oviducts / Q000187:drug effects; D012733:Sex Differentiation / Q000187:drug effects; D013737:Testis / Q000187:drug effects; D014874:Water Pollutants, Chemical / Q000633:toxicity*; D014981:Xenopus / Q000196:embryology / Q000502:physiology*
chemical_list: D047628:Estrogen Receptor alpha; D004967:Estrogens; D014874:Water Pollutants, Chemical; D004997:Ethinyl Estradiol