Significance of AHR nuclear translocation sequence in 2,3,7,8-tetrachlorodibenzo-p-dioxin-induced cPLA₂α activation and hydronephrosis

The aryl hydrocarbon receptor (AHR) plays a major role in 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced toxicity phenotypes. TCDD bound to AHR elicits both genomic action in which target genes are transcriptionally upregulated and nongenomic action in which cytosolic phospholipase A₂α (cPLA₂α) is rapidly activated. However, how either of these actions, separately or in combination, induces toxicity phenotypes is largely unknown. In this study, we used AHR^nls/nls mice as a model in which AHR was mutated to lack nuclear translocation sequence (NLS), and AHR^d/− mice as the corresponding control. Using this model, we studied TCDD-induced alterations in cPLA₂α activation and related factors because of the pivotal roles of cPLA₂α both in AHR's nongenomic action and in regulation of causative genes of TCDD-induced hydronephrosis. Dams were orally administered TCDD at a dose of 300 µg/kg body weight on postnatal day 1, and pups subsequently exposed to TCDD via milk were examined for gene expression on PND 7 and for histological changes on PND 14. The activation of the AHR genomic action and hydronephrosis onset were observed in the control group but not in the AHR^nls/nls group. An ex vivo experiment using peritoneal macrophages exposed to 100 nM TCDD resulted in rapid activation of cPLA₂α, an indicator of the nongenomic action, only in the control group but not in the AHR^nls/nls group. These results indicated that an NLS is required for the AHR's genomic and nongenomic actions.