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The aryl hydrocarbon receptor (Ahr), a member of the large basic helix-loop-helix (bHLH) and PAS homology domain superfamily, is a highly conserved transcriptional regulator involved in mammalian development. In the present study, a murine metanephros organ culture system was employed to evaluate the role of the Ahr signaling in nephrogenesis in vitro. Ahr and Wilms' tumor suppressor (wt1) mRNAs were detected by in situ hybridization and RT-PCR during the course of renal development. Treatment with 3 microM BaP, a hydrocarbon ligand of Ahr, inhibited glomerulogenesis and branching morphogenesis of metanephric kidneys. Deficits in the epithelialization of mesenchymal cells were evidenced by inhibition of the formation of podocyte foot processes and glomerular basement membranes. Hydrocarbon treatment markedly induced -KTS wt1 splice variants, although total wt1 mRNA levels remained unchanged. A significant decrease in total WT1 protein was observed by both immunocytochemistry and Western analysis in cultures challenged with BaP compared to controls. Comparison of metanephric cultures from Ahr+/+ and Ahr-/- mice showed that Ahr is involved in kidney development, and required for BaP-induced deficits in nephrogenesis. These results indicate that ligand activation of Ahr signaling disrupts nephrogenesis in vitro, and that this response involves modulation of wt1 alternative splicing and post-transcriptional control.

Ligand-activated Ahr signaling leads to disruption of nephrogenesis and altered Wilms' tumor suppressor mRNA splicing