In utero and lactational exposure of the male rat to 2,3,7,8-tetrachlorodibenzo-p-dioxin impairs prostate development. 1. Effects on gene expression.

摘要

In utero and lactational 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) exposure decreases rat prostate weight without decreasing circulating androgen concentrations. Because one mechanism by which TCDD is thought to cause toxicity is by aryl hydrocarbon receptor (AhR)-mediated alterations in gene transcription, the goals of this study were to determine whether the developing prostate expresses the AhR and its dimerization partner, the AhR nuclear translocator (ARNT); to determine whether in utero and lactational TCDD exposure is capable of directly activating gene transcription in the developing prostate; and to identify prostatic mRNAs that exhibit altered abundance in response to in utero and lactational TCDD exposure. Pregnant Holtzman rats were administered TCDD (1.0 microgram/kg po) or vehicle on Gestation Day (GD) 15, and male offspring were euthanized between Postnatal Days (PNDs) 1 and 63. Using reverse transcriptase-polymerase chain reaction (RT-PCR), mRNAs encoding the AhR and ARNT were detected in both ventral and dorsolateral prostates from control animals throughout postnatal development. ARNT protein was expressed in the majority of stromal nuclei early in development, whereas ARNT expression in the prostate epithelium was initially cytoplasmic but became nuclear as development progressed. GD 15 TCDD exposure increased cytochrome P4501A1 (CYP1A1) mRNA and protein in whole prostates between PNDs 7 and 21. In these TCDD-exposed animals, CYP1A1 protein was localized to the epithelium. In order to define other genes in the developing prostate that might be regulated by TCDD at the level of mRNA, RNA samples from PND 21 whole prostates from control and TCDD-exposed animals were compared using mRNA differential display. Although no growth-regulatory candidates were identified using this screening technique, a ventral prostate-specific, androgen-regulated mRNA (20-kDa protein) was identified that seemed to be downregulated by TCDD exposure. Northern blot analysis confirmed this decrease at PND 21 and further showed that the downregulation was transient. Similar results were obtained for four additional androgen-regulated prostatic mRNAs (prostatic binding protein [PBP], Royal Winnipeg Ballet [RWB], probasin, and dorsal protein-1 [DP-1]), all of which are markers of a differentiated ductal epithelium. In contrast, TCDD exposure of adult male rats (25 micrograms TCDD/kg, 24 h) greatly induced CYP1A1 mRNA without affecting the abundance of prostate-specific, androgen-regulated mRNAs. These results suggest that the transient decreases in androgen-regulated prostatic mRNA abundance observed in response to in utero and lactational TCDD exposure were probably not the result of direct action of the activated AhR on these genes but instead were reflective of a TCDD-induced delay in prostate development.