Impact of Fetal Exposure to Endocrine Disrupting Chemical Mixtures on FOXA3 Gene and Protein Expression in Adult Rat Testes

Perinatal exposure to endocrine disrupting chemicals (EDCs) has been shown to affect male reproductive functions. However, the effects on male reproduction of exposure to EDC mixtures at doses relevant to humans have not been fully characterized. In previous studies, we found that in utero exposure to mixtures of the plasticizer di(2-ethylhexyl) phthalate (DEHP) and the soy-based phytoestrogen genistein (Gen) induced abnormal testis development in rats. In the present study, we investigated the molecular basis of these effects in adult testes from the offspring of pregnant SD rats gavaged with corn oil or Gen + DEHP mixtures at 0.1 or 10 mg/kg/day. Testicular transcriptomes were determined by microarray and RNA-seq analyses. A protein analysis was performed on paraffin and frozen testis sections, mainly by immunofluorescence. The transcription factor forkhead box protein 3 (FOXA3), a key regulator of Leydig cell function, was identified as the most significantly downregulated gene in testes from rats exposed in utero to Gen + DEHP mixtures. FOXA3 protein levels were decreased in testicular interstitium at a dose previously found to reduce testosterone levels, suggesting a primary effect of fetal exposure to Gen + DEHP on adult Leydig cells, rather than on spermatids and Sertoli cells, also expressing FOXA3. Thus, FOXA3 downregulation in adult testes following fetal exposure to Gen + DEHP may contribute to adverse male reproductive outcomes.

[1]  J. Mendiola,et al.  Temporal trends in sperm count: a systematic review and meta-regression analysis of samples collected globally in the 20th and 21st centuries. , 2022, Human reproduction update.

[2]  M. Skinner,et al.  Environmental induced transgenerational inheritance impacts systems epigenetics in disease etiology , 2022, Scientific Reports.

[3]  G. Adamkiewicz,et al.  Phthalate and novel plasticizer concentrations in food items from U.S. fast food chains: a preliminary analysis , 2021, Journal of Exposure Science & Environmental Epidemiology.

[4]  Huatao Chen,et al.  Circadian regulation of apolipoprotein gene expression affects testosterone production in mouse testis. , 2021, Theriogenology.

[5]  Hui Li,et al.  Environmental exposures, fetal testis development and function: phthalates and beyond. , 2021, Reproduction.

[6]  V. Papadopoulos,et al.  Cholesterol-binding translocator protein TSPO regulates steatosis and bile acid synthesis in nonalcoholic fatty liver disease , 2021, iScience.

[7]  V. Papadopoulos,et al.  Impact of endocrine-disrupting chemicals on steroidogenesis and consequences on testicular function , 2021, Molecular and Cellular Endocrinology.

[8]  W. Dhillo,et al.  Male infertility due to testicular disorders. , 2020, The Journal of clinical endocrinology and metabolism.

[9]  S. Grimm,et al.  TMEFF2: A Transmembrane Proteoglycan with Multifaceted Actions in Cancer and Disease , 2020, Cancers.

[10]  M. Lupien,et al.  Pioneer of prostate cancer: past, present and the future of FOXA1 , 2020, Protein & cell.

[11]  C. Foresta,et al.  Effects of endocrine disruptors on fetal testis development, male puberty, and transition age , 2020, Endocrine.

[12]  Annie Boisvert,et al.  In utero exposure to low doses of genistein and di‐(2‐ethylhexyl) phthalate (DEHP) alters innate immune cells in neonatal and adult rat testes , 2020, Andrology.

[13]  C. Jayasena,et al.  Endocrine‐disrupting chemicals and male reproductive health , 2020, Reproductive medicine and biology.

[14]  S. Lacagnina.  The Developmental Origins of Health and Disease (DOHaD) , 2020, American journal of lifestyle medicine.

[15]  Qiqi Zhu,et al.  Dehydroepiandrosterone and Its CYP7B1 Metabolite 7α-Hydroxydehydroepiandrosterone Regulates 11β-Hydroxysteroid Dehydrogenase 1 Directions in Rat Leydig Cells , 2020, Frontiers in Endocrinology.

[16]  Y. Shima.  Development of fetal and adult Leydig cells , 2019, Reproductive medicine and biology.

[17]  P. Roumaud,et al.  Transcriptomic analysis of overexpressed SOX4 and SOX8 in TM4 Sertoli cells with emphasis on cell-to-cell interactions. , 2019, Biochemical and biophysical research communications.

[18]  Qiqi Zhu,et al.  Endocrine disruptors of inhibiting testicular 3β-hydroxysteroid dehydrogenase. , 2019, Chemico-biological interactions.

[19]  S. Majumdar,et al.  Regulation of Hippo pathway components by FSH in testis. , 2019, Reproductive biology.

[20]  A. Giwercman,et al.  Endocrine disruptors and testicular function. , 2018, Metabolism: clinical and experimental.

[21]  C. Krausz,et al.  Genetics of male infertility , 2018, Nature Reviews Urology.

[22]  H. Tournaye,et al.  The diagnosis of male infertility: an analysis of the evidence to support the development of global WHO guidance—challenges and future research opportunities , 2017, Human reproduction update.

[23]  E. Larriba,et al.  Chronic low-dose exposure to a mixture of environmental endocrine disruptors induces microRNAs/isomiRs deregulation in mouse concomitant with intratesticular estradiol reduction , 2017, Scientific Reports.

[24]  Haojie Huang,et al.  CXCL13 is androgen-responsive and involved in androgen induced prostate cancer cell migration and invasion. , 2017, Oncotarget.

[25]  F. Bergeron,et al.  FOXA3 Is Expressed in Multiple Cell Lineages in the Mouse Testis and Regulates Pdgfra Expression in Leydig Cells , 2017, Endocrinology.

[26]  S. Wołczyński,et al.  Endocrine-disrupting chemicals—Mechanisms of action on male reproductive system , 2017, Toxicology and industrial health.

[27]  P. Jenardhanan,et al.  Effect of environmental contaminants on spermatogenesis. , 2016, Seminars in cell & developmental biology.

[28]  M. Culty,et al.  Regulation of Translocator Protein 18 kDa (TSPO) Expression in Rat and Human Male Germ Cells , 2016, International journal of molecular sciences.

[29]  C. Langoya,et al.  Household air pollution and childhood pneumonia in South Sudan: will clean cooking stoves reduce the incidence and mortality? , 2016 .

[30]  R. Nusse,et al.  Paracrine Wnt/β-catenin signaling mediates proliferation of undifferentiated spermatogonia in the adult mouse testis , 2016, Proceedings of the National Academy of Sciences.

[31]  F. Brimo,et al.  Changes in the expression profiles of claudins during gonocyte differentiation and in seminomas , 2016, Andrology.

[32]  M. Griswold.  Spermatogenesis: The Commitment to Meiosis. , 2016, Physiological reviews.

[33]  A C Gore,et al.  EDC-2: The Endocrine Society's Second Scientific Statement on Endocrine-Disrupting Chemicals. , 2015, Endocrine reviews.

[34]  Steven Jones,et al.  In Utero Exposure to Di-(2-Ethylhexyl) Phthalate Induces Testicular Effects in Neonatal Rats That Are Antagonized by Genistein Cotreatment1 , 2015, Biology of reproduction.

[35]  K. Loveland,et al.  Interactome of the negative regulator of nuclear import BRCA1-binding protein 2 , 2015, Scientific Reports.

[36]  V. Papadopoulos,et al.  Fetal origin of endocrine dysfunction in the adult: The phthalate model , 2013, The Journal of Steroid Biochemistry and Molecular Biology.

[37]  K. Tsutsui,et al.  Localization of gonadotropin-releasing hormone (GnRH), gonadotropin-inhibitory hormone (GnIH), kisspeptin and GnRH receptor and their possible roles in testicular activities from birth to senescence in mice. , 2012, Journal of experimental zoology. Part A, Ecological genetics and physiology.

[38]  S. Wing,et al.  Expression of the Ubiquitin Proteasome System in Neonatal Rat Gonocytes and Spermatogonia: Role in Gonocyte Differentiation1 , 2012, Biology of reproduction.

[39]  J. Cravedi,et al.  Effect of mono-(2-ethylhexyl) phthalate on human and mouse fetal testis: In vitro and in vivo approaches. , 2012, Toxicology and applied pharmacology.

[40]  W. Foster,et al.  Phytoestrogens in Human Pregnancy , 2012, Obstetrics and gynecology international.

[41]  Kim Boekelheide,et al.  Human Fetal Testis Xenografts Are Resistant to Phthalate-Induced Endocrine Disruption , 2012, Environmental health perspectives.

[42]  R. Sharpe,et al.  Do phthalates affect steroidogenesis by the human fetal testis? Exposure of human fetal testis xenografts to di-n-butyl phthalate. , 2012, The Journal of clinical endocrinology and metabolism.

[43]  B. Benayoun,et al.  Forkhead transcription factors: key players in health and disease. , 2011, Trends in genetics : TIG.

[44]  V. Papadopoulos,et al.  Mitochondrial protein import and the genesis of steroidogenic mitochondria , 2011, Molecular and Cellular Endocrinology.

[45]  Yan Wang,et al.  Changes in MAPK pathway in neonatal and adult testis following fetal estrogen exposure and effects on rat testicular cells , 2009, Microscopy research and technique.

[46]  R. Behringer,et al.  Fetal Leydig Cell Origin and Development , 2009, Sexual Development.

[47]  V. Papadopoulos,et al.  In utero exposure to di-(2-ethylhexyl) phthalate exerts both short-term and long-lasting suppressive effects on testosterone production in the rat. , 2008, Biology of reproduction.

[48]  R. Sharpe,et al.  Testicular dysgenesis syndrome: mechanistic insights and potential new downstream effects. , 2008, Fertility and sterility.

[49]  Phillip P. Le,et al.  Impaired male fertility and atrophy of seminiferous tubules caused by haploinsufficiency for Foxa3. , 2007, Developmental biology.

[50]  Sue Marty,et al.  NTP-CERHR expert panel report on the reproductive and developmental toxicity of genistein. , 2006, Birth defects research. Part B, Developmental and reproductive toxicology.

[51]  Katherine Shea,et al.  NTP-CERHR Expert Panel Update on the Reproductive and Developmental Toxicity of di(2-ethylhexyl) phthalate. , 2006, Reproductive toxicology.

[52]  D. Nutt,et al.  Translocator protein (18kDa): new nomenclature for the peripheral-type benzodiazepine receptor based on its structure and molecular function. , 2006, Trends in pharmacological sciences.

[53]  C. Chambers,et al.  NTP-CERHR expert panel report on the reproductive and developmental toxicity of soy formula. , 2006, Birth defects research. Part B, Developmental and reproductive toxicology.

[54]  Yan Wang,et al.  Prenatal Estrogen Exposure Differentially Affects Estrogen Receptor-Associated Proteins in Rat Testis Gonocytes1 , 2004, Biology of reproduction.

[55]  G. Presta,et al.  In utero exposure to di-(2-ethylhexyl)phthalate and duration of human pregnancy. , 2003, Environmental health perspectives.

[56]  Yan Wang,et al.  Prenatal Exposure to Estrogenic Compounds Alters the Expression Pattern of Platelet-Derived Growth Factor Receptors α and β in Neonatal Rat Testis: Identification of Gonocytes as Targets of Estrogen Exposure1 , 2003, Biology of reproduction.

[57]  J. Mohler,et al.  Identification of differentially expressed genes associated with androgen‐independent growth of prostate cancer , 2002, The Prostate.

[58]  W. Foster,et al.  Detection of phytoestrogens in samples of second trimester human amniotic fluid. , 2002, Toxicology letters.