title: The interaction of Sertoli and Leydig cells in the testicular toxicity of tri-o-cresyl phosphate.
abstract: Previous studies have shown that after dosing with tri-o-cresyl phosphate (TOCP), the testis contains more active intermediate (saligenin cyclic-o-tolyl phosphate; SCOTP) than do other organs or blood. SCOTP is produced by a cytochrome P450-dependent reaction, and the Sertoli cells, although containing little P450, are the testicular cells that show the first signs of damage after TOCP administration. The present studies evaluated (i) whether testicular Leydig cell production of SCOTP might explain the elevated testicular concentration of SCOTP, (ii) if this production affected testosterone secretion, and (iii) if Sertoli cells cocultured over TOCP-exposed Leydig cells would show effects similar to those found after SCOTP exposure of Sertoli cells in vitro, indicating a cell interaction. Previous data showed that a target enzyme for SCOTP in Sertoli cells, nonspecific esterase (NSE), was inhibited by exposure in vitro to SCOTP, but not to TOCP. In the present experiments, HPLC analysis identified SCOTP in media from Leydig cells cultured with radiolabeled TOCP, demonstrating activation. TOCP addition to Leydig cells decreased testosterone output after stimulation with hCG, an effect that was replicated by subsequent in vivo experiments. Addition of various intermediates in the testosterone biosynthesis pathway indicated that both mitochondrial- and microsomal-based steps in the pathway were affected. Collectively, these data indicate that Leydig cells can activate TOCP. To model whether this activation might affect Sertoli cells in vivo, Sertoli cells were plated in culture-well inserts suspended above (cocultured with) isolated Leydig cells in the presence of TOCP. Sertoli NSE activity was diminished, while remaining unchanged when cultured in the presence of TOCP but without Leydig cells, or over Leydig cells alone. These results show that the Leydig cells in the testis are capable of activating TOCP to SCOTP, and that this can produce effects in Sertoli cells. This in situ activation of TOCP to SCOTP may help explain why the testis contains high concentrations of SCOTP after in vivo dosing with TOCP, and why the testis is a target organ for TOCP toxicity.
authors.affiliation: Developmental and Reproductive Toxicology Group, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709.
authors.forename: R E
authors.initials: RE
authors.lastname: Chapin
authors.forename: J L
authors.initials: JL
authors.lastname: Phelps
authors.forename: S G
authors.initials: SG
authors.lastname: Somkuti
authors.forename: J J
authors.initials: JJ
authors.lastname: Heindel
authors.forename: L T
authors.initials: LT
authors.lastname: Burka
mesh_terms: D000735:Androstenedione / Q000494:pharmacology; D000818:Animals; D043182:Carboxylesterase; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism; D002450:Cell Communication / Q000187:drug effects; D002478:Cells, Cultured; D002784:Cholesterol / Q000494:pharmacology; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D003408:Cresols / Q000633:toxicity*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D066298:In Vitro Techniques; D007985:Leydig Cells / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D009943:Organophosphorus Compounds / Q000097:blood / Q000378:metabolism; D011284:Pregnenolone / Q000494:pharmacology; D011374:Progesterone / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D012708:Sertoli Cells / Q000187:drug effects; D013737:Testis / Q000187:drug effects* / Q000378:metabolism; D013739:Testosterone / Q000378:metabolism; D014317:Tritolyl Phosphates / Q000633:toxicity*
chemical_list: D006063:Chorionic Gonadotropin; D003408:Cresols; D009943:Organophosphorus Compounds; D014317:Tritolyl Phosphates; C000516:2-(2-cresyl)-4H-1-3-2-benzodioxaphosphorin-2-oxide; D013739:Testosterone; D000735:Androstenedione; D011374:Progesterone; D011284:Pregnenolone; D002784:Cholesterol; D002265:Carboxylic Ester Hydrolases; D043182:Carboxylesterase; C025541:tri-o-cresyl phosphate; D004121:Dimethyl Sulfoxide