title: Mechanisms of toxicity of di(2-ethylhexyl) phthalate on the reproductive health of male zebrafish.
abstract: Phthalates are ubiquitous in the aquatic environment and are known to adversely affect male reproductive health in mammals through interactions with multiple receptor systems. However, little is known about the risks they pose to fish. This project investigated the effects of di(2-ethylhexyl) phthalate (DEHP), the most commonly used phthalate, on the reproductive health of male zebrafish (Danio rerio). Males were treated with 0.5, 50 and 5000 mg DEHP kg(-1) (body weight) for a period of 10 days via intraperitoneal injection. The effects of the exposure were assessed by analysing fertilisation success, testis histology, sperm DNA integrity and transcript profiles of the liver and testis. A significant increase in the hepatosomatic index and levels of hepatic vitellogenin transcript were observed following exposure to 5000 mg DEHP kg(-1). Exposure to 5000 mg DEHP kg(-1) also resulted in a reduction in fertilisation success of oocytes spawned by untreated females. However, survival and development of the resulting embryos were unaffected by all treatments, and no evidence of DEHP-induced sperm DNA damage was observed. Exposure to 50 and 5000 mg DEHP kg(-1) caused alterations in the proportion of germ cells at specific stages of spermatogenesis in the testis, including a reduction in the proportion of spermatozoa and an increase in the proportion of spermatocytes, suggesting that DEHP may inhibit the progression of meiosis. In parallel, exposure to 5000 mg DEHP kg(-1) increased the levels of two peroxisome proliferator-activated receptor (PPAR) responsive genes (acyl-coenzyme A oxidase 1 (acox1) and enoyl-coenzyme A, hydratase/3-hydroxyacyl coenzyme A dehydrogenase (ehhadh). These data demonstrated that exposure to high concentrations of DEHP disrupts spermatogenesis in adult zebrafish with a consequent decrease in their ability to fertilise oocytes spawned by untreated females. Furthermore, our data suggest that the adverse effects caused by exposure to DEHP are likely to occur preferentially via PPAR signalling pathways in the testis and oestrogen signalling pathways in the liver. We found no evidence of adverse effects on zebrafish reproductive health following exposure to the concentrations occurring in most aquatic systems, indicating that DEHP alone may not be a causative agent of the reproductive abnormalities seen in wildlife, at least as a result of short-term exposures.
authors.affiliation: Hatherly Laboratories, School of Biosciences, University of Exeter, Exeter, Devon EX4 4PS, UK.
authors.forename: Tamsyn M
authors.initials: TM
authors.lastname: Uren-Webster
authors.forename: Ceri
authors.initials: C
authors.lastname: Lewis
authors.forename: Amy L
authors.initials: AL
authors.lastname: Filby
authors.forename: Gregory C
authors.initials: GC
authors.lastname: Paull
authors.forename: Eduarda M
authors.initials: EM
authors.lastname: Santos
mesh_terms: D000818:Animals; D004249:DNA Damage; D004051:Diethylhexyl Phthalate / Q000633:toxicity*; D004625:Embryo, Nonmammalian / Q000187:drug effects; D005260:Female; D005306:Fertilization / Q000187:drug effects; D020869:Gene Expression Profiling; D008297:Male; D012098:Reproduction / Q000187:drug effects*; D013091:Spermatogenesis / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects; D014874:Water Pollutants, Chemical / Q000633:toxicity*; D015027:Zebrafish / Q000502:physiology*
chemical_list: D014874:Water Pollutants, Chemical; D004051:Diethylhexyl Phthalate