Aryl hydrocarbon receptor ligands increase ABC transporter activity and protein expression in killifish (Fundulus heteroclitus) renal proximal tubules.

Many widespread and persistent organic pollutants, for example, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and some polychlorinated biphenyls, activate the aryl hydrocarbon receptor (AhR) causing it to translocate to the cell nucleus where it transactivates target genes, increasing expression of a number of xenobiotic metabolizing enzymes as well as some transporters. AhR's ability to target transporters within the kidney is essentially unexplored. We show here that exposing isolated killifish (Fundulus heteroclitus) renal proximal tubules to micromolar ß-naphthoflavone (BNF) or nanomolar TCDD roughly doubled the transport activity of Multidrug resistance-associated proteins Mrp2 and Mrp4, P-glycoprotein (P-gp) and Breast cancer resistance protein (Bcrp), all ATP-driven xenobiotic efflux pumps and critical determinants of renal xenobiotic excretion. These effects were abolished by actinomycin D and cycloheximide and by the AhR antagonist, α-naphthoflavone, indicating that increased transport activity was dependent on transcription and translation as well as ligand binding to AhR. Quantitative immunostaining of renal tubules exposed to BNF and TCDD showed increased luminal membrane expression of Mrp2, Mrp4, P-gp and Bcrp. Thus, in these renal tubules, the four ABC transporters are targets of AhR action.

Characterization of efflux transport proteins of the human choroid plexus papilloma cell line HIBCPP, a functional in vitro model of the blood-cerebrospinal fluid barrier.

PURPOSE: To characterize the human choroid plexus (CP) papilloma cell line HIBCPP with respect to ABC export protein expression and function in order to evaluate its use as an in vitro model to study carrier-mediated transport processes at the CP. METHODS: Expression profiles of ABC transporters were studied by quantitative real-time PCR and Western Blot analysis. Functionality of transporters was investigated by means of uptake experiments and permeation studies carried out on permeable filter systems. In addition, immunohistochemistry served to study localization of ABCC1 and ABCC4. RESULTS: Both qPCR and Western Blot revealed that ABC transporters known to be expressed in CP are also expressed in HIBCPP cells. Immunohistochemistry confirmed basolateral expression of ABCC1. Functionality of ABCC1, ABCC4, ABCB1 and ABCG2 could be shown in uptake assays. CONCLUSIONS: Altogether, the HIBCPP cells promise to be a functional and relevant in vitro tool to investigate transport processes at the blood-cerebrospinal fluid barrier.