title: Prepubertal Di-n-Butyl Phthalate Exposure Alters Sertoli and Leydig Cell Function and Lowers Bone Density in Adult Male Mice.
abstract: Phthalate exposure impairs testis development and function; however, whether phthalates affect nonreproductive functions is not well understood. To investigate this, C57BL/6J mice were fed 1-500 mg di-n-butyl phthalate (DBP) in corn oil, or vehicle only, daily from 4 to 14 days, after which tissues were collected (prepubertal study). Another group was fed 1-500 mg/kg·d DBP from 4 to 21 days and then maintained untreated until 8 weeks for determination of adult consequences of prepubertal exposure. Bones were assessed by microcomputed tomography and dual-energy X-ray absorptiometry and T by RIA. DBP exposure decreased prepubertal femur length, marrow volume, and mean moment of inertia. Adult animals exposed prepubertally to low DBP doses had lower bone mineral content and bone mineral density and less lean tissue mass than vehicle-treated animals. Altered dynamics of the emerging Leydig population were found in 14-day-old animals fed 100-500 mg/kg·d DBP. Adult mice had variable testicular T and serum T and LH concentrations after prepubertal exposure and a dose-dependent reduction in cytochrome p450, family 11, subfamily A, polypeptide 1. Insulin-like 3 was detected in Sertoli cells of adult mice administered the highest dose of 500 mg/kg·d DBP prepubertally, a finding supported by the induction of insulin-like 3 expression in TM4 cells exposed to 50 μM, but not 5 μM, DBP. We propose that low-dose DBP exposure is detrimental to bone but that normal bone mineral density/bone mineral content after high-dose DBP exposure reflects changes in testicular somatic cells that confer protection to bones. These findings will fuel concerns that low-dose DBP exposure impacts health beyond the reproductive axis.
authors.affiliation: Priority Research Centres for Reproductive Science and Chemical Biology, School of Environmental and Life Sciences (A.B., C.I.), and School of Mathematical and Physical Sciences (N.C.), Faculty of Science and Information Technology, University of Newcastle, Callaghan, New South Wales 2308, Australia; St Vincent's Institute of Medical Research (R.W.J., I.J.P., N.A.S.) and Department of Medicine at St. Vincent's Hospital (R.W.J., I.J.P., N.A.S.), The University of Melbourne, Fitzroy, Victoria 3065, Australia; Departments of Biochemistry and Molecular Biology (H.G., K.L.L.), Anatomy and Developmental Biology (S.C.M., K.L.L.), and Molecular Translational Sciences (K.L.L.), Monash University, and Hudson Institute of Medical Research (K.L.L., M.P.H.), Clayton, Victoria 3800, Australia; and Faculty of Science, Health, Education, and Engineering (C.I.), School of Health and Sport Sciences, University of the Sunshine Coast, Sippy Downs, Queensland 4556, Australia.
authors.forename: Amanda
authors.initials: A
authors.lastname: Bielanowicz
authors.affiliation: Priority Research Centres for Reproductive Science and Chemical Biology, School of Environmental and Life Sciences (A.B., C.I.), and School of Mathematical and Physical Sciences (N.C.), Faculty of Science and Information Technology, University of Newcastle, Callaghan, New South Wales 2308, Australia; St Vincent's Institute of Medical Research (R.W.J., I.J.P., N.A.S.) and Department of Medicine at St. Vincent's Hospital (R.W.J., I.J.P., N.A.S.), The University of Melbourne, Fitzroy, Victoria 3065, Australia; Departments of Biochemistry and Molecular Biology (H.G., K.L.L.), Anatomy and Developmental Biology (S.C.M., K.L.L.), and Molecular Translational Sciences (K.L.L.), Monash University, and Hudson Institute of Medical Research (K.L.L., M.P.H.), Clayton, Victoria 3800, Australia; and Faculty of Science, Health, Education, and Engineering (C.I.), School of Health and Sport Sciences, University of the Sunshine Coast, Sippy Downs, Queensland 4556, Australia.
authors.forename: Rachelle W
authors.initials: RW
authors.lastname: Johnson
authors.affiliation: Priority Research Centres for Reproductive Science and Chemical Biology, School of Environmental and Life Sciences (A.B., C.I.), and School of Mathematical and Physical Sciences (N.C.), Faculty of Science and Information Technology, University of Newcastle, Callaghan, New South Wales 2308, Australia; St Vincent's Institute of Medical Research (R.W.J., I.J.P., N.A.S.) and Department of Medicine at St. Vincent's Hospital (R.W.J., I.J.P., N.A.S.), The University of Melbourne, Fitzroy, Victoria 3065, Australia; Departments of Biochemistry and Molecular Biology (H.G., K.L.L.), Anatomy and Developmental Biology (S.C.M., K.L.L.), and Molecular Translational Sciences (K.L.L.), Monash University, and Hudson Institute of Medical Research (K.L.L., M.P.H.), Clayton, Victoria 3800, Australia; and Faculty of Science, Health, Education, and Engineering (C.I.), School of Health and Sport Sciences, University of the Sunshine Coast, Sippy Downs, Queensland 4556, Australia.
authors.forename: Hoey
authors.initials: H
authors.lastname: Goh
authors.affiliation: Priority Research Centres for Reproductive Science and Chemical Biology, School of Environmental and Life Sciences (A.B., C.I.), and School of Mathematical and Physical Sciences (N.C.), Faculty of Science and Information Technology, University of Newcastle, Callaghan, New South Wales 2308, Australia; St Vincent's Institute of Medical Research (R.W.J., I.J.P., N.A.S.) and Department of Medicine at St. Vincent's Hospital (R.W.J., I.J.P., N.A.S.), The University of Melbourne, Fitzroy, Victoria 3065, Australia; Departments of Biochemistry and Molecular Biology (H.G., K.L.L.), Anatomy and Developmental Biology (S.C.M., K.L.L.), and Molecular Translational Sciences (K.L.L.), Monash University, and Hudson Institute of Medical Research (K.L.L., M.P.H.), Clayton, Victoria 3800, Australia; and Faculty of Science, Health, Education, and Engineering (C.I.), School of Health and Sport Sciences, University of the Sunshine Coast, Sippy Downs, Queensland 4556, Australia.
authors.forename: Sarah C
authors.initials: SC
authors.lastname: Moody
authors.affiliation: Priority Research Centres for Reproductive Science and Chemical Biology, School of Environmental and Life Sciences (A.B., C.I.), and School of Mathematical and Physical Sciences (N.C.), Faculty of Science and Information Technology, University of Newcastle, Callaghan, New South Wales 2308, Australia; St Vincent's Institute of Medical Research (R.W.J., I.J.P., N.A.S.) and Department of Medicine at St. Vincent's Hospital (R.W.J., I.J.P., N.A.S.), The University of Melbourne, Fitzroy, Victoria 3065, Australia; Departments of Biochemistry and Molecular Biology (H.G., K.L.L.), Anatomy and Developmental Biology (S.C.M., K.L.L.), and Molecular Translational Sciences (K.L.L.), Monash University, and Hudson Institute of Medical Research (K.L.L., M.P.H.), Clayton, Victoria 3800, Australia; and Faculty of Science, Health, Education, and Engineering (C.I.), School of Health and Sport Sciences, University of the Sunshine Coast, Sippy Downs, Queensland 4556, Australia.
authors.forename: Ingrid J
authors.initials: IJ
authors.lastname: Poulton
authors.affiliation: Priority Research Centres for Reproductive Science and Chemical Biology, School of Environmental and Life Sciences (A.B., C.I.), and School of Mathematical and Physical Sciences (N.C.), Faculty of Science and Information Technology, University of Newcastle, Callaghan, New South Wales 2308, Australia; St Vincent's Institute of Medical Research (R.W.J., I.J.P., N.A.S.) and Department of Medicine at St. Vincent's Hospital (R.W.J., I.J.P., N.A.S.), The University of Melbourne, Fitzroy, Victoria 3065, Australia; Departments of Biochemistry and Molecular Biology (H.G., K.L.L.), Anatomy and Developmental Biology (S.C.M., K.L.L.), and Molecular Translational Sciences (K.L.L.), Monash University, and Hudson Institute of Medical Research (K.L.L., M.P.H.), Clayton, Victoria 3800, Australia; and Faculty of Science, Health, Education, and Engineering (C.I.), School of Health and Sport Sciences, University of the Sunshine Coast, Sippy Downs, Queensland 4556, Australia.
authors.forename: Nic
authors.initials: N
authors.lastname: Croce
authors.affiliation: Priority Research Centres for Reproductive Science and Chemical Biology, School of Environmental and Life Sciences (A.B., C.I.), and School of Mathematical and Physical Sciences (N.C.), Faculty of Science and Information Technology, University of Newcastle, Callaghan, New South Wales 2308, Australia; St Vincent's Institute of Medical Research (R.W.J., I.J.P., N.A.S.) and Department of Medicine at St. Vincent's Hospital (R.W.J., I.J.P., N.A.S.), The University of Melbourne, Fitzroy, Victoria 3065, Australia; Departments of Biochemistry and Molecular Biology (H.G., K.L.L.), Anatomy and Developmental Biology (S.C.M., K.L.L.), and Molecular Translational Sciences (K.L.L.), Monash University, and Hudson Institute of Medical Research (K.L.L., M.P.H.), Clayton, Victoria 3800, Australia; and Faculty of Science, Health, Education, and Engineering (C.I.), School of Health and Sport Sciences, University of the Sunshine Coast, Sippy Downs, Queensland 4556, Australia.
authors.forename: Kate L
authors.initials: KL
authors.lastname: Loveland
authors.affiliation: Priority Research Centres for Reproductive Science and Chemical Biology, School of Environmental and Life Sciences (A.B., C.I.), and School of Mathematical and Physical Sciences (N.C.), Faculty of Science and Information Technology, University of Newcastle, Callaghan, New South Wales 2308, Australia; St Vincent's Institute of Medical Research (R.W.J., I.J.P., N.A.S.) and Department of Medicine at St. Vincent's Hospital (R.W.J., I.J.P., N.A.S.), The University of Melbourne, Fitzroy, Victoria 3065, Australia; Departments of Biochemistry and Molecular Biology (H.G., K.L.L.), Anatomy and Developmental Biology (S.C.M., K.L.L.), and Molecular Translational Sciences (K.L.L.), Monash University, and Hudson Institute of Medical Research (K.L.L., M.P.H.), Clayton, Victoria 3800, Australia; and Faculty of Science, Health, Education, and Engineering (C.I.), School of Health and Sport Sciences, University of the Sunshine Coast, Sippy Downs, Queensland 4556, Australia.
authors.forename: Mark P
authors.initials: MP
authors.lastname: Hedger
authors.affiliation: Priority Research Centres for Reproductive Science and Chemical Biology, School of Environmental and Life Sciences (A.B., C.I.), and School of Mathematical and Physical Sciences (N.C.), Faculty of Science and Information Technology, University of Newcastle, Callaghan, New South Wales 2308, Australia; St Vincent's Institute of Medical Research (R.W.J., I.J.P., N.A.S.) and Department of Medicine at St. Vincent's Hospital (R.W.J., I.J.P., N.A.S.), The University of Melbourne, Fitzroy, Victoria 3065, Australia; Departments of Biochemistry and Molecular Biology (H.G., K.L.L.), Anatomy and Developmental Biology (S.C.M., K.L.L.), and Molecular Translational Sciences (K.L.L.), Monash University, and Hudson Institute of Medical Research (K.L.L., M.P.H.), Clayton, Victoria 3800, Australia; and Faculty of Science, Health, Education, and Engineering (C.I.), School of Health and Sport Sciences, University of the Sunshine Coast, Sippy Downs, Queensland 4556, Australia.
authors.forename: Natalie A
authors.initials: NA
authors.lastname: Sims
authors.affiliation: Priority Research Centres for Reproductive Science and Chemical Biology, School of Environmental and Life Sciences (A.B., C.I.), and School of Mathematical and Physical Sciences (N.C.), Faculty of Science and Information Technology, University of Newcastle, Callaghan, New South Wales 2308, Australia; St Vincent's Institute of Medical Research (R.W.J., I.J.P., N.A.S.) and Department of Medicine at St. Vincent's Hospital (R.W.J., I.J.P., N.A.S.), The University of Melbourne, Fitzroy, Victoria 3065, Australia; Departments of Biochemistry and Molecular Biology (H.G., K.L.L.), Anatomy and Developmental Biology (S.C.M., K.L.L.), and Molecular Translational Sciences (K.L.L.), Monash University, and Hudson Institute of Medical Research (K.L.L., M.P.H.), Clayton, Victoria 3800, Australia; and Faculty of Science, Health, Education, and Engineering (C.I.), School of Health and Sport Sciences, University of the Sunshine Coast, Sippy Downs, Queensland 4556, Australia.
authors.forename: Catherine
authors.initials: C
authors.lastname: Itman
mesh_terms: D015502:Absorptiometry, Photon; D000818:Animals; D015519:Bone Density / Q000187:drug effects*; D003993:Dibutyl Phthalate / Q000494:pharmacology*; D005269:Femur / Q000000981:diagnostic imaging / Q000187:drug effects*; D007985:Leydig Cells / Q000187:drug effects* / Q000378:metabolism; D007986:Luteinizing Hormone / Q000097:blood; D008297:Male; D051379:Mice; D010968:Plasticizers / Q000494:pharmacology; D012708:Sertoli Cells / Q000187:drug effects* / Q000378:metabolism; D013737:Testis / Q000187:drug effects / Q000378:metabolism; D013739:Testosterone / Q000378:metabolism; D055114:X-Ray Microtomography
chemical_list: D010968:Plasticizers; D003993:Dibutyl Phthalate; D013739:Testosterone; D007986:Luteinizing Hormone