Aryl Hydrocarbon Receptor Activation in Genital Tubercle, Palate, and Other Embryonic Tissues in 2,3,7,8-Tetrachlorodibenzo-p-Dioxin-ResponsivelacZMice

Abstract

The aryl hydrocarbon receptor (AhR) is a ligand-dependent transcription factor that mediates the toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and related halogenated aromatic hydrocarbons. Although the normal function and endogenous ligand for this receptor are not known, it is thought to have a role in growth regulation processes. The AhR has been found in both adult and certain developing tissues, and AhR agonists like the environmental contaminant TCDD cause a number of developmental anomalies. We sought to determine whether the AhR is directly activated to a transcriptionally functional form in tissues known to be adversely affected by AhR agonist exposure. To this end, a transgenic mouse model was developed that could be used to indicate the temporal and spatial context of transcriptionally active AhR following agonist exposurein vivo.A synthetic promoter containing two dioxin-responsive elements (DREs) and a minimal TATA box was strongly induced by TCDD in transfected cells when linked to thelacZor luciferase reporter gene. Transgenic mice harboring thelacZconstruct had TCDD-inducible β-galactosidase activity in tissues following adult andin uteroexposure. EmbryoniclacZexpression was induced in hard and soft palates, genital tubercle, certain facial regions, shoulder, as well as other tissues byin uteroexposure to 30 μg TCDD/kg at Gestational Day 13. The most intense reporter response was observed in the genital tubercle. Histopathology of the palate and tubercle demonstrated the reporter gene activity to be both cell- and region-specific. This is the first publication to correlate reported TCDD-elicited toxicity (e.g., cleft palate in mice) with TCDD-dependent AhR activation. These data indicate the ability of TCDD to initiate a signal transduction process leading to a transcriptionally active AhR in these tissues, thereby identifying potential targets of dioxin-induced toxicity during development. Weak activation of the reporter gene was consistently observed only in the genital tubercle in the absence of exogenous inducer. This indicates minimal or no endogenous AhR activators at the developmental stage examined. This mouse model will prove useful for both the examination of the endogenous role of the AhR in proliferation or differentiation and of the developmental targets of dioxin-like compounds.