title: Endocrine disruptors differentially target ATP-binding cassette transporters in the blood-testis barrier and affect Leydig cell testosterone secretion in vitro.
abstract: Endocrine-disrupting chemicals (EDCs) are considered to cause testicular toxicity primarily via interference with steroid hormone function. Alternatively, EDCs could possibly exert their effects by interaction with ATP-binding cassette (ABC) transporters that are expressed in the blood-testis barrier. In this study, we investigated the effects of bisphenol A (BPA), tetrabromobisphenol A (TBBPA), bis(2-ethylhexyl) phthalate, mono(2-ethylhexyl) phthalate, perfluorooctanoic acid (PFOA), and perfluorooctanesulfonic acid (PFOS) on breast cancer resistance protein (BCRP), multidrug resistance proteins 1 and 4 (MRP1 and MRP4), and P-glycoprotein (P-gp) using membrane vesicles overexpressing these transporters. BPA solely inhibited BCRP activity, whereas TBBPA, PFOA, and PFOS inhibited all transporters tested. No effect was observed for the phthalates. Using transporter-overexpressing Madin-Darby canine kidney cells, we show that BPA and PFOA, but not TBBPA, are transported by BCRP, whereas none of the compounds were transported by P-gp. To investigate the toxicological implications of these findings, testosterone secretion and expression of steroidogenic genes were determined in murine Leydig (MA-10) cells upon exposure to the selected EDCs. Only BPA and TBBPA concentration dependently increased testosterone secretion by MA-10 cells to 6- and 46-fold of control levels, respectively. Inhibition of the Mrp's by MK-571 completely blocked testosterone secretion elicited by TBBPA, which could not be explained by coinciding changes in expression of steroidogenic genes. Therefore, we hypothesize that transporter-mediated efflux of testosterone precursors out of MA-10 cells is inhibited by TBBPA resulting in higher availability for testosterone production. Our data show the toxicological and clinical relevance of ABC transporters in EDC risk assessment related to testicular toxicity.
authors.affiliation: * Department of Pharmacology and Toxicology, Radboud University Nijmegen Medical Centre, 6500 HB Nijmegen, The Netherlands;
authors.forename: Anita C A
authors.initials: AC
authors.lastname: Dankers
authors.forename: Maarke J E
authors.initials: MJ
authors.lastname: Roelofs
authors.forename: Aldert H
authors.initials: AH
authors.lastname: Piersma
authors.forename: Fred C G J
authors.initials: FC
authors.lastname: Sweep
authors.forename: Frans G M
authors.initials: FG
authors.lastname: Russel
authors.forename: Martin
authors.initials: M
authors.lastname: van den Berg
authors.forename: Majorie B M
authors.initials: MB
authors.lastname: van Duursen
authors.forename: Rosalinde
authors.initials: R
authors.lastname: Masereeuw
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000818:Animals; D001814:Blood-Testis Barrier / Q000187:drug effects*; D002478:Cells, Cultured; D004285:Dogs; D052244:Endocrine Disruptors / Q000633:toxicity*; D057809:HEK293 Cells; D006801:Humans; D007985:Leydig Cells / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D051379:Mice; D013739:Testosterone / Q000378:metabolism*
chemical_list: D018528:ATP-Binding Cassette Transporters; D052244:Endocrine Disruptors; D013739:Testosterone
keywords: ABC transporters; MA-10 mouse Leydig cells; blood-testis barrier; endocrine disrupting chemicals (EDCs); steroidogenesis.