Novel cDNA sequences of aryl hydrocarbon receptors and gene expression in turtles (Chrysemys picta and Pseudemys scripta) exposed to different environments.

Reproductive changes have been observed in painted turtles from a site with known contamination located on Cape Cod, MA, USA. We hypothesize that these changes are caused by exposure to endocrine-disrupting compounds and that genes involved in reproduction are affected. The aryl hydrocarbon receptor (AHR) is an orphan receptor that is activated by environmental contaminants. AHR mRNA was measured in turtles exposed to soil collected from a contaminated site. Adult turtles were trapped from the study site (Moody Pond, MP) or a reference site and exposed to laboratory environments containing soil from either site. The red-eared slider was used to assess neonatal exposure to soil and water from the sites. The environmental exposures occurred over a 13-month period. Juveniles showed an age-dependent increase in brain AHR1. Juvenile turtles exposed to the MP environment had elevated gonadal AHR1. Adult turtles exposed to the MP environment showed significantly decreased brain AHR2. The painted turtle AHR is the first complete reptile AHR cDNA sequence. Phylogenetic analysis of the painted turtle AHR showed that it clusters with other AHR2s. Partial AHR1 and partial AHR2 cDNA sequences were cloned from the red-eared slider. MEME analysis identified 18 motifs in the turtle AHRs, showing high conservation between motifs that overlapped functional regions in both AHR isoforms.

Cloning of estrogen receptor alpha and aromatase cDNAs and gene expression in turtles (Chrysemys picta and Pseudemys scripta) exposed to different environments.

Reproductive changes have been observed in painted turtles (Chrysemys picta) from a site with known contamination located on Cape Cod, MA, USA. We hypothesize that these changes are caused by exposure to endocrine-disrupting compounds and that genes playing a significant role in reproduction are affected. cDNA sequences were determined for estrogen receptor alpha and aromatase in the painted turtle. These genes were measured in our study animals using quantitative PCR. Adult turtles were trapped from our study site (Moody Pond, MP) or a reference site (Washburn Pond) and exposed to laboratory environments containing soil from either site. The red-eared slider (Pseudemys scripta), a pond turtle closely related to the painted turtle, was used to assess neonatal exposure to soil and water from the sites. Our results show an increase in hepatic estrogen receptor, which suggests exposure to estrogenic contaminants. Female turtles from MP appear to have a long-term effect on hepatic ER. Other findings were apparent age-dependent differences in expression of aromatase and ER in the brains of neonate and year-old juvenile turtles. Phylogenetic analyses of the cDNA sequences further support the hypothesis that turtles are in a sister clade to birds and crocodilians.