Man is not a big rat: concerns with traditional human risk assessment of phthalates based on their anti-androgenic effects observed in the rat foetus

Basic and Clinical Andrology

René Habert¹  Gabriel Livera²  Virginie Rouiller-Fabre² 

[1] Stem Cells and Radiation Unit, LDG / SCSR / iRCM / DSV, Centre CEA, BP6, F-92265 Fontenay aux Roses, France

[2] INSERM, Unité 967, F-92265 Fontenay aux Roses, France

关键词: Testosterone;    Testis;    Foetus;    Reproduction;    Development;    Toxicity test;    Risk assessment;    Human health;    Masculinization;    Leydig cells;    Phthalates;    Endocrine disruptors;   

Others  :  1083957
DOI  :  10.1186/2051-4190-24-14

 received in 2014-06-24, accepted in 2014-08-15,  发布年份 2014

【 摘 要 】

Phthalates provide one of the most documented example evidencing how much we must be cautious when using the traditional paradigm based on extrapolation of experimental data from rodent studies for human health risk assessment of endocrine disruptors (EDs). Since foetal testis is known as one of the most sensitive targets of EDs, phthalate risk assessment is routinely based on the capacity of such compounds to decrease testosterone production by the testis or to impair masculinization in the rat during foetal life. In this paper, the well-established inhibiting effects of phthalates of the foetal Leydig cells function in the rat are briefly reviewed. Then, data obtained in humans and other species are carefully analysed. Already in January 2009, using the organotypic culture system named Fetal Testis Assay (FeTA) that we developed, we reported that phthalates might not affect testosterone production in human foetal testes. Several recent experimental studies using xenografts confirm the absence of detectable anti-androgenic effect of phthalates in the human foetal testes. Epidemiological studies led to contradictory results. Altogether, these findings suggest that phthalates effects on foetal Leydig cells are largely species-specific. Consequently, the phthalate threshold doses that disturb foetal steroidogenesis in rat testes and that are presently used to define the acceptable daily intake levels for human health protection must be questioned. This does not mean that phthalates are safe because these compounds have many deleterious effects upon germ cell development that may be common to the different studied species including human. More generally, the identification of common molecular, cellular or/and phenotypic targets in rat and human testes should precede the choice of the toxicological endpoint in rat to accurately assess the safety threshold of any ED in humans.

【 授权许可】

   
2014 Habert et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

【 参考文献 】

• [1]Habert R, Muczynski V, Grisin T, Moison D, Messiaen S, Frydman R, Benachi A, Delbes G, Lambrot R, Lehraiki A, N’Tumba-Byn T, Guerquin MJ, Levacher C, Rouiller-Fabre V, Livera G: Concerns about the widespread use of rodent models for human risk assessments of endocrine disruptors. Reproduction 2014, 147:R119-129.
• [2]Habert R, Picon R: Attempts for identification of a chorionic gonadotrophin-like bioactivity in the rat placenta which stimulates the testosterone secretion of the fetal testis in vitro. Biol Neonate 1990, 58:24-31.
• [3]Habert R, Lejeune H, Saez JM: Origin, differentiation and regulation of fetal and adult Leydig cells. Mol Cell Endocrinol 2001, 179:47-74.
• [4]Leridon H, Slama R: The impact of a decline in fecundity and of pregnancy postponement on final number of children and demand for assisted reproduction technology. Hum Reprod 2008, 23:1312-1319.
• [5]Main KM, Skakkebaek NE, Virtanen HE, Toppari J: Genital anomalies in boys and the environment. Best Pract Res Clin Endocrinol Metab 2010, 24:279-289.
• [6]Sharpe RM, Irvine DS: How strong is the evidence of a link between environmental chemicals and adverse effects on human reproductive health? BMJ 2004, 328:447-451.
• [7]Toppari J, Larsen JC, Christiansen P, Giwercman A, Grandjean P, Guillette LJ Jr, Jegou B, Jensen TK, Jouannet P, Keiding N, Leffers H, McLachlan JA, Meyer O, Muller J, Rajpert-De Meyts E, Scheike T, Sharpe R, Sumpter J, Skakkebaek NE: Male reproductive health and environmental xenoestrogens. Environ Health Perspect 1996, 104(Suppl 4):741-803.
• [8]Le Moal J, Rolland M, Goria S, Wagner V, De Crouy-Chanel P, Rigou A, De Mouzon J, Royere D: Semen quality trends in French regions are consistent with a global change in environmental exposure. Reproduction 2014, 147:567-574.
• [9]Skakkebaek NE, Rajpert-De Meyts E, Main KM: Testicular dysgenesis syndrome: an increasingly common developmental disorder with environmental aspects. Hum Reprod 2001, 16:972-978.
• [10]Olesen IA, Sonne SB, Hoei-Hansen CE, Rajpert-De Meyts E, Skakkebaek NE: Environment, testicular dysgenesis and carcinoma in situ testis. Best Pract Res Clin Endocrinol Metab 2007, 21:462-478.
• [11]Delbes G, Levacher C, Habert R: Estrogen effects on fetal and neonatal testicular development. Reproduction 2006, 132:527-538.
• [12]Toppari J, Virtanen HE, Main KM, Skakkebaek NE: Cryptorchidism and hypospadias as a sign of testicular dysgenesis syndrome (TDS): environmental connection. Birth Defects Res Part A Clin Mol Teratol 2010, 88:910-919.
• [13]Adamo C, Antignac J, Auger J, Balaguer P, Bourc’his D, Bujan L, Chevrier C, Cotinot C, Cravedi J, Laudet V, Livera G, Slama R: Reproduction et Environnement. Paris: INSERM edn; 2011.
• [14]Vandenberg LN, Colborn T, Hayes TB, Heindel JJ, Jacobs DR Jr, Lee DH, Myers JP, Shioda T, Soto AM, vom Saal FS, Welshons WV, Zoeller RT: Regulatory decisions on endocrine disrupting chemicals should be based on the principles of endocrinology. Reprod Toxicol 2013, 38:1-15.
• [15]Giwercman A, Giwercman YL: Environmental factors and testicular function. Best Pract Res Clin Endocrinol Metab 2011, 25:391-402.
• [16]Habert R, Muczynski V, Lehraiki A, Lambrot R, Lecureuil C, Levacher C, Coffigny H, Pairault C, Moison D, Frydman R, Rouiller-Fabre V: Adverse effects of endocrine disruptors on the foetal testis development: focus on the phthalates. Folia Histochem Cyto 2009, 47:S67-74.
• [17]Sharpe RM, Skakkebaek NE: Are oestrogens involved in falling sperm counts and disorders of the male reproductive tract? Lancet 1993, 341:1392-1395.
• [18]Juul A, Almstrup K, Andersson AM, Jensen TK, Jorgensen N, Main KM, Meyts ER, Toppari J, Skakkebaek NE: Possible fetal determinants of male infertility. Nat Rev Endocrinol 2014.
• [19]Svechnikov K, Stukenborg JB, Savchuck I, Soder O: Similar causes of various reproductive disorders in early life. Asian J Androl 2014, 16:50-59.
• [20]Sharpe RM, Skakkebaek NE: Testicular dysgenesis syndrome: mechanistic insights and potential new downstream effects. Fertil Steril 2008, 89:e33-38.
• [21]Gray LE Jr, Ostby J, Furr J, Price M, Veeramachaneni DN, Parks L: Perinatal exposure to the phthalates DEHP, BBP, and DINP, but not DEP, DMP, or DOTP, alters sexual differentiation of the male rat. Toxicol Sci 2000, 58:350-365.
• [22]Higuchi TT, Palmer JS, Gray LE Jr, Veeramachaneni DN: Effects of dibutyl phthalate in male rabbits following in utero, adolescent, or postpubertal exposure. Toxicol Sci 2003, 72:301-313.
• [23]Mahood IK, Scott HM, Brown R, Hallmark N, Walker M, Sharpe RM: In utero exposure to di(n-butyl) phthalate and testicular dysgenesis: comparison of fetal and adult end points and their dose sensitivity. Environ Health Perspect 2007, 115(Suppl 1):55-61.
• [24]Mylchreest E, Foster PM: DBP exerts its antiandrogenic activity by indirectly interfering with androgen signaling pathways. Toxicol Appl Pharmacol 2000, 168:174-175.
• [25]Parks LG, Ostby JS, Lambright CR, Abbott BD, Klinefelter GR, Barlow NJ, Gray LE Jr: The plasticizer diethylhexyl phthalate induces malformations by decreasing fetal testosterone synthesis during sexual differentiation in the male rat. Toxicol Sci 2000, 58:339-349.
• [26]Wine RN, Li LH, Barnes LH, Gulati DK, Chapin RE: Reproductive toxicity of di-n-butylphthalate in a continuous breeding protocol in Sprague–Dawley rats. Environ Health Perspect 1997, 105:102-107.
• [27]Becker K, Seiwert M, Angerer J, Heger W, Koch HM, Nagorka R, Rosskamp E, Schluter C, Seifert B, Ullrich D: DEHP metabolites in urine of children and DEHP in house dust. Int J Hyg Environ Health 2004, 207:409-417.
• [28]Koch HM, Becker K, Wittassek M, Seiwert M, Angerer J, Kolossa-Gehring M: Di-n-butylphthalate and butylbenzylphthalate - urinary metabolite levels and estimated daily intakes: pilot study for the German Environmental Survey on children. J Expo Sci Environ Epidemiol 2007, 17:378-387.
• [29]Wormuth M, Scheringer M, Vollenweider M, Hungerbuhler K: What are the sources of exposure to eight frequently used phthalic acid esters in Europeans? Risk Anal 2006, 26:803-824.
• [30]Latini G: Monitoring phthalate exposure in humans. Clin Chim Acta 2005, 361:20-29.
• [31]Saravanabhavan G, Murray J: Human biological monitoring of diisononyl phthalate and diisodecyl phthalate: a review. J Environ Public Health 2012, 2012:810501.
• [32]Koch HM, Bolt HM, Preuss R, Angerer J: New metabolites of di(2-ethylhexyl)phthalate (DEHP) in human urine and serum after single oral doses of deuterium-labelled DEHP. Arch Toxicol 2005, 79:367-376.
• [33]Koch HM, Drexler H, Angerer J: An estimation of the daily intake of di(2-ethylhexyl)phthalate (DEHP) and other phthalates in the general population. Int J Hyg Environ Health 2003, 206:77-83.
• [34]Marsee K, Woodruff TJ, Axelrad DA, Calafat AM, Swan SH: Estimated daily phthalate exposures in a population of mothers of male infants exhibiting reduced anogenital distance. Environ Health Perspect 2006, 114:805-809.
• [35]Latini G, De Felice C, Presta G, Del Vecchio A, Paris I, Ruggieri F, Mazzeo P: In utero exposure to di-(2-ethylhexyl)phthalate and duration of human pregnancy. Environ Health Perspect 2003, 111:1783-1785.
• [36]Silva MJ, Barr DB, Reidy JA, Malek NA, Hodge CC, Caudill SP, Brock JW, Needham LL, Calafat AM: Urinary levels of seven phthalate metabolites in the U.S. population from the National Health and Nutrition Examination Survey (NHANES) 1999–2000. Environ Health Perspect 2004, 112:331-338.
• [37]Swan SH, Main KM, Liu F, Stewart SL, Kruse RL, Calafat AM, Mao CS, Redmon JB, Ternand CL, Sullivan S, Teague JL, Study for Future Families Research T: Decrease in anogenital distance among male infants with prenatal phthalate exposure. Environ Health Perspect 2005, 113:1056-1061.
• [38]Main KM, Mortensen GK, Kaleva MM, Boisen KA, Damgaard IN, Chellakooty M, Schmidt IM, Suomi AM, Virtanen HE, Petersen DV, Andersson AM, Toppari J, Skakkebaek NE: Human breast milk contamination with phthalates and alterations of endogenous reproductive hormones in infants three months of age. Environ Health Perspect 2006, 114:270-276.
• [39]Hogberg J, Hanberg A, Berglund M, Skerfving S, Remberger M, Calafat AM, Filipsson AF, Jansson B, Johansson N, Appelgren M, Hakansson H: Phthalate diesters and their metabolites in human breast milk, blood or serum, and urine as biomarkers of exposure in vulnerable populations. Environ Health Perspect 2008, 116:334-339.
• [40]Huang PC, Kuo PL, Chou YY, Lin SJ, Lee CC: Association between prenatal exposure to phthalates and the health of newborns. Environ Int 2009, 35:14-20.
• [41]Suzuki Y, Yoshinaga J, Mizumoto Y, Serizawa S, Shiraishi H: Foetal exposure to phthalate esters and anogenital distance in male newborns. Int J Androl 2012, 35:236-244.
• [42]Bustamante-Montes LP, Hernandez-Valero MA, Flores-Pimentel D, Garcia-Fabila M, Amaya-Chavez A, Barr DB, Borja-Aburto VH: Prenatal exposure to phthalates is associated with decreased anogenital distance and penile size in male newborns. J Dev Orig Health Dis 2013, 4:300-306.
• [43]Scott HM, Mason JI, Sharpe RM: Steroidogenesis in the fetal testis and its susceptibility to disruption by exogenous compounds. Endocr Rev 2009, 30:883-925.
• [44]O’Shaughnessy PJ, Fowler PA: Endocrinology of the mammalian fetal testis. Reproduction 2011, 141:37-46.
• [45]Svechnikov K, Landreh L, Weisser J, Izzo G, Colon E, Svechnikova I, Soder O: Origin, development and regulation of human Leydig cells. Horm Res Paediatr 2010, 73:93-101.
• [46]Tapanainen J, Kellokumpu-Lehtinen P, Pelliniemi L, Huhtaniemi I: Age-related changes in endogenous steroids of human fetal testis during early and midpregnancy. J Clin Endocrinol Metab 1981, 52:98-102.
• [47]Lambrot R, Livera G, Coffigny H, Pairault C, Frydman R, Habert R, Rouiller-Fabre V: A new method for toxicity assays on human and mouse fetal testis. Biochimie 2006, 88:1831-1835.
• [48]Habert R, Picon R: Testosterone, dihydrotestosterone and estradiol-17 beta levels in maternal and fetal plasma and in fetal testes in the rat. J Steroid Biochem 1984, 21:193-198.
• [49]Tapanainen J, Kuopio T, Pelliniemi LJ, Huhtaniemi I: Rat testicular endogenous steroids and number of Leydig cells between the fetal period and sexual maturity. Biol Reprod 1984, 31:1027-1035.
• [50]Kerr JB, Knell CM: The fate of fetal Leydig cells during the development of the fetal and postnatal rat testis. Development 1988, 103:535-544.
• [51]Habert R, Brignaschi P: Developmental changes in in vitro testosterone production by dispersed Leydig cells during fetal life in rats. Arch Androl 1991, 27:65-71.
• [52]Huhtaniemi I: Endocrine function and regulation of the fetal and neonatal testis. Int J Dev Biol 1989, 33:117-123.
• [53]Habert R, Veniard B, Brignaschi P, Gangnerau MN, Picon R: Absence of development of late steroidogenic lesions in rat testis during the end of fetal life. Arch Androl 1989, 22:41-48.
• [54]Habert R, Picon R: Control of testicular steroidogenesis in foetal rat: effect of decapitation on testosterone and plasma luteinizing hormone-like activity. Acta Endocrinol (Copenh) 1982, 99:466-473.
• [55]Migrenne S, Pairault C, Racine C, Livera G, Geloso A, Habert R: Luteinizing hormone-dependent activity and luteinizing hormone-independent differentiation of rat fetal Leydig cells. Mol Cell Endocrinol 2001, 172:193-202.
• [56]Habert R: In vivo acute testicular testosterone response to injection of luteinizing hormone in the rat fetus. Acta Endocrinol (Copenh) 1993, 128:268-273.
• [57]Borday C, Merlet J, Racine C, Habert R: Expression and localization of aromatase during fetal mouse testis development. Basic Clin Androl 2013, 23:13.
• [58]Migrenne S, Moreau E, Pakarinen P, Dierich A, Merlet J, Habert R, Racine C: Mouse testis development and function are differently regulated by follicle-stimulating hormone receptors signaling during fetal and prepubertal life. PLoS ONE 2012, 7:e53257.
• [59]Svechnikov K, Izzo G, Landreh L, Weisser J, Soder O: Endocrine disruptors and Leydig cell function. J Biomed Biotechnol 2010, 2010:Article ID684504.
• [60]Martinez-Arguelles DB, Campioli E, Culty M, Zirkin BR, Papadopoulos V: Fetal origin of endocrine dysfunction in the adult: the phthalate model. J Steroid Biochem Mol Biol 2013, 137:5-17.
• [61]Barlow NJ, Foster PM: Pathogenesis of male reproductive tract lesions from gestation through adulthood following in utero exposure to Di(n-butyl) phthalate. Toxicol Pathol 2003, 31:397-410.
• [62]Mahood IK, Hallmark N, McKinnell C, Walker M, Fisher JS, Sharpe RM: Abnormal Leydig Cell aggregation in the fetal testis of rats exposed to di (n-butyl) phthalate and its possible role in testicular dysgenesis. Endocrinology 2005, 146:613-623.
• [63]Hallmark N, Walker M, McKinnell C, Mahood IK, Scott H, Bayne R, Coutts S, Anderson RA, Greig I, Morris K, Sharpe RM: Effects of monobutyl and di(n-butyl) phthalate in vitro on steroidogenesis and Leydig cell aggregation in fetal testis explants from the rat: comparison with effects in vivo in the fetal rat and neonatal marmoset and in vitro in the human. Environ Health Perspect 2007, 115:390-396.
• [64]Fisher JS, Macpherson S, Marchetti N, Sharpe RM: Human 'testicular dysgenesis syndrome’: a possible model using in-utero exposure of the rat to dibutyl phthalate. Hum Reprod 2003, 18:1383-1394.
• [65]Lehmann KP, Phillips S, Sar M, Foster PM, Gaido KW: Dose-dependent alterations in gene expression and testosterone synthesis in the fetal testes of male rats exposed to di (n-butyl) phthalate. Toxicol Sci 2004, 81:60-68.
• [66]Howdeshell KL, Rider CV, Wilson VS, Gray LE Jr: Mechanisms of action of phthalate esters, individually and in combination, to induce abnormal reproductive development in male laboratory rats. Environ Res 2008, 108:168-176.
• [67]Saillenfait AM, Sabate JP, Robert A, Rouiller-Fabre V, Roudot AC, Moison D, Denis F: Dose-dependent alterations in gene expression and testosterone production in fetal rat testis after exposure to di-n-hexyl phthalate. J Appl Toxicol 2013, 33:1027-1035.
• [68]Thompson CJ, Ross SM, Gaido KW: Di(n-butyl) phthalate impairs cholesterol transport and steroidogenesis in the fetal rat testis through a rapid and reversible mechanism. Endocrinology 2004, 145:1227-1237.
• [69]Borch J, Metzdorff SB, Vinggaard AM, Brokken L, Dalgaard M: Mechanisms underlying the anti-androgenic effects of diethylhexyl phthalate in fetal rat testis. Toxicology 2006, 223:144-155.
• [70]Culty M, Thuillier R, Li W, Wang Y, Martinez-Arguelles DB, Benjamin CG, Triantafilou KM, Zirkin BR, Papadopoulos V: In utero exposure to di-(2-ethylhexyl) phthalate exerts both short-term and long-lasting suppressive effects on testosterone production in the rat. Biol Reprod 2008, 78:1018-1028.
• [71]van den Driesche S, Walker M, McKinnell C, Scott HM, Eddie SL, Mitchell RT, Seckl JR, Drake AJ, Smith LB, Anderson RA, Sharpe RM: Proposed role for COUP-TFII in regulating fetal Leydig cell steroidogenesis, perturbation of which leads to masculinization disorders in rodents. PLoS ONE 2012, 7:e37064.
• [72]Anand-Ivell R, Ivell R: Insulin-like factor 3 as a monitor of endocrine disruption. Reproduction 2014, 147:R87-95.
• [73]Wilson VS, Lambright C, Furr J, Ostby J, Wood C, Held G, Gray LE Jr: Phthalate ester-induced gubernacular lesions are associated with reduced insl3 gene expression in the fetal rat testis. Toxicol Lett 2004, 146:207-215.
• [74]McKinnell C, Sharpe RM, Mahood K, Hallmark N, Scott H, Ivell R, Staub C, Jegou B, Haag F, Koch-Nolte F, Hartung S: Expression of insulin-like factor 3 protein in the rat testis during fetal and postnatal development and in relation to cryptorchidism induced by in utero exposure to di (n-Butyl) phthalate. Endocrinology 2005, 146:4536-4544.
• [75]Howdeshell KL, Wilson VS, Furr J, Lambright CR, Rider CV, Blystone CR, Hotchkiss AK, Gray LE Jr: A mixture of five phthalate esters inhibits fetal testicular testosterone production in the Sprague–Dawley rat in a cumulative, dose-additive manner. Toxicol Sci 2008, 105:153-165.
• [76]Jost A, Vigier B, Prepin J, Perchellet JP: Studies on sex differentiation in mammals. Recent Prog Horm Res 1973, 29:1-41.
• [77]Jost A: Hormonal factors in the sex differentiation of the mammalian foetus. Philos T Roy Soc B 1970, 259:119-130.
• [78]Welsh M, Saunders PT, Fisken M, Scott HM, Hutchison GR, Smith LB, Sharpe RM: Identification in rats of a programming window for reproductive tract masculinization, disruption of which leads to hypospadias and cryptorchidism. J Clin Invest 2008, 118:1479-1490.
• [79]Macleod DJ, Sharpe RM, Welsh M, Fisken M, Scott HM, Hutchison GR, Drake AJ, van den Driesche S: Androgen action in the masculinization programming window and development of male reproductive organs. Int J Androl 2010, 33:279-287.
• [80]Gray LE, Ostby J, Furr J, Wolf CJ, Lambright C, Parks L, Veeramachaneni DN, Wilson V, Price M, Hotchkiss A, Orlando E, Guillette L: Effects of environmental antiandrogens on reproductive development in experimental animals. Hum Reprod Update 2001, 7:248-264.
• [81]Foster PM, Mylchreest E, Gaido KW, Sar M: Effects of phthalate esters on the developing reproductive tract of male rats. Hum Reprod Update 2001, 7:231-235.
• [82]Fisher JS: Environmental anti-androgens and male reproductive health: focus on phthalates and testicular dysgenesis syndrome. Reproduction 2004, 127:305-315.
• [83]Foster PM: Disruption of reproductive development in male rat offspring following in utero exposure to phthalate esters. Int J Androl 2006, 29:140-147. discussion 181–145
• [84]Gray LE Jr, Wilson VS, Stoker T, Lambright C, Furr J, Noriega N, Howdeshell K, Ankley GT, Guillette L: Adverse effects of environmental antiandrogens and androgens on reproductive development in mammals. Int J Androl 2006, 29:96-104. discussion 105–108
• [85]Toppari J, Virtanen H, Skakkebaek NE, Main KM: Environmental effects on hormonal regulation of testicular descent. J Steroid Biochem Mol Biol 2006, 102:184-186.
• [86]Ge RS, Chen GR, Tanrikut C, Hardy MP: Phthalate ester toxicity in Leydig cells: developmental timing and dosage considerations. Reprod Toxicol 2007, 23:366-373.
• [87]Hu GX, Lian QQ, Ge RS, Hardy DO, Li XK: Phthalate-induced testicular dysgenesis syndrome: Leydig cell influence. Trends Endocrinol Metab 2009, 20:139-145.
• [88]Struve MF, Gaido KW, Hensley JB, Lehmann KP, Ross SM, Sochaski MA, Willson GA, Dorman DC: Reproductive toxicity and pharmacokinetics of di-n-butyl phthalate (DBP) following dietary exposure of pregnant rats. Birth Defects Res B Dev Reprod Toxicol 2009, 86:345-354.
• [89]Lee KY, Shibutani M, Takagi H, Kato N, Takigami S, Uneyama C, Hirose M: Diverse developmental toxicity of di-n-butyl phthalate in both sexes of rat offspring after maternal exposure during the period from late gestation through lactation. Toxicology 2004, 203:221-238.
• [90]Clewell RA, Thomas A, Willson G, Creasy DM, Andersen ME: A dose response study to assess effects after dietary administration of diisononyl phthalate (DINP) in gestation and lactation on male rat sexual development. Reprod Toxicol 2013, 35:70-80.
• [91]Gray LE Jr, Barlow NJ, Howdeshell KL, Ostby JS, Furr JR, Gray CL: Transgenerational effects of Di (2-ethylhexyl) phthalate in the male CRL:CD(SD) rat: added value of assessing multiple offspring per litter. Toxicol Sci 2009, 110:411-425.
• [92]Christiansen S, Boberg J, Axelstad M, Dalgaard M, Vinggaard AM, Metzdorff SB, Hass U: Low-dose perinatal exposure to di(2-ethylhexyl) phthalate induces anti-androgenic effects in male rats. Reprod Toxicol 2010, 30:313-321.
• [93]Liu K, Lehmann KP, Sar M, Young SS, Gaido KW: Gene expression profiling following in utero exposure to phthalate esters reveals new gene targets in the etiology of testicular dysgenesis. Biol Reprod 2005, 73:180-192.
• [94]Witorsch RJ, Thomas JA: Personal care products and endocrine disruption: a critical review of the literature. Crit Rev Toxicol 2010, 40(Suppl 3):1-30.
• [95]Johnson KJ, Heger NE, Boekelheide K: Of mice and men (and rats): phthalate-induced fetal testis endocrine disruption is species-dependent. Toxicol Sci 2012, 129:235-248.
• [96]Veeramachaneni DN, Klinefelter GR: Phthalate-induced pathology in the foetal testis involves more than decreased testosterone production. Reproduction 2014, 147:435-442.
• [97]Swan SH: Environmental phthalate exposure in relation to reproductive outcomes and other health endpoints in humans. Environ Res 2008, 108:177-184.
• [98]Choi H, Kim J, Im Y, Lee S, Kim Y: The association between some endocrine disruptors and hypospadias in biological samples. J Environ Sci Health A Tox 2012, 47:2173-2179.
• [99]Ormond G, Nieuwenhuijsen MJ, Nelson P, Toledano MB, Iszatt N, Geneletti S, Elliott P: Endocrine disruptors in the workplace, hair spray, folate supplementation, and risk of hypospadias: case–control study. Environ Health Perspect 2009, 117:303-307.
• [100]Teitelbaum SL, Britton JA, Calafat AM, Ye X, Silva MJ, Reidy JA, Galvez MP, Brenner BL, Wolff MS: Temporal variability in urinary concentrations of phthalate metabolites, phytoestrogens and phenols among minority children in the United States. Environ Res 2008, 106:257-269.
• [101]Sathyanarayana S, Barrett E, Butts S, Wang C, Swan SH: Phthalate exposure and reproductive hormone concentrations in pregnancy. Reproduction 2014, 147:401-409.
• [102]N’Tumba-Byn T, Moison D, Lacroix M, Lecureuil C, Lesage L, Prud’homme SM, Pozzi-Gaudin S, Frydman R, Benachi A, Livera G, Rouiller-Fabre V, Habert R: Differential effects of bisphenol A and diethylstilbestrol on human, rat and mouse fetal leydig cell function. PLoS ONE 2012, 7:e51579.
• [103]Habert R, Devif I, Gangnerau MN, Lecerf L: Ontogenesis of the in vitro response of rat testis to gonadotropin-releasing hormone. Mol Cell Endocrinol 1991, 82:199-206.
• [104]Lambrot R, Coffigny H, Pairault C, Donnadieu AC, Frydman R, Habert R, Rouiller-Fabre V: Use of organ culture to study the human fetal testis development: effect of retinoic acid. J Clin Endocrinol Metab 2006, 91:2696-2703.
• [105]Lecerf L, Rouiller-Fabre V, Levacher C, Gautier C, Saez JM, Habert R: Stimulatory effect of follicle-stimulating hormone on basal and luteinizing hormone-stimulated testosterone secretions by the fetal rat testis in vitro. Endocrinology 1993, 133:2313-2318.
• [106]Olaso R, Pairault C, Boulogne B, Durand P, Habert R: Transforming growth factor beta1 and beta2 reduce the number of gonocytes by increasing apoptosis. Endocrinology 1998, 139:733-740.
• [107]Livera G, Rouiller-Fabre V, Durand P, Habert R: Multiple effects of retinoids on the development of Sertoli, germ, and Leydig cells of fetal and neonatal rat testis in culture. Biol Reprod 2000, 62:1303-1314.
• [108]Livera G, Delbes G, Pairault C, Rouiller-Fabre V, Habert R: Organotypic culture, a powerful model for studying rat and mouse fetal testis development. Cell Tissue Res 2006, 324:507-521.
• [109]Lambrot R, Muczynski V, Lecureuil C, Angenard G, Coffigny H, Pairault C, Moison D, Frydman R, Habert R, Rouiller-Fabre V: Phthalates impair germ cell development in the human fetal testis in vitro without change in testosterone production. Environ Health Perspect 2009, 117:32-37.
• [110]Muczynski V, Cravedi JP, Lehraiki A, Levacher C, Moison D, Lecureuil C, Messiaen S, Perdu E, Frydman R, Habert R, Rouiller-Fabre V: Effect of mono-(2-ethylhexyl) phthalate on human and mouse fetal testis: In vitro and in vivo approaches. Toxicol Appl Pharmacol 2012, 261:97-104.
• [111]Muczynski V, Lecureuil C, Messiaen S, Guerquin MJ, N’Tumba-Byn T, Moison D, Hodroj W, Benjelloun H, Baijer J, Livera G, Frydman R, Benachi A, Habert R, Rouiller-Fabre V: Cellular and molecular effect of MEHP Involving LXRalpha in human fetal testis and ovary. PLoS ONE 2012, 7:e48266.
• [112]Delbes G, Duquenne C, Szenker J, Taccoen J, Habert R, Levacher C: Developmental changes in testicular sensitivity to estrogens throughout fetal and neonatal life. Toxicol Sci 2007, 99:234-243.
• [113]Mitchell RT, Saunders PT, Childs AJ, Cassidy-Kojima C, Anderson RA, Wallace WH, Kelnar CJ, Sharpe RM: Xenografting of human fetal testis tissue: a new approach to study fetal testis development and germ cell differentiation. Hum Reprod 2010, 25:2405-2414.
• [114]Mitchell RT, Childs AJ, Anderson RA, van den Driesche S, Saunders PT, McKinnell C, Wallace WH, Kelnar CJ, Sharpe RM: Do phthalates affect steroidogenesis by the human fetal testis? Exposure of human fetal testis xenografts to di-n-butyl phthalate. J Clin Endocrinol Metab 2012, 97:E341-348.
• [115]Heger NE, Hall SJ, Sandrof MA, McDonnell EV, Hensley JB, McDowell EN, Martin KA, Gaido KW, Johnson KJ, Boekelheide K: Human fetal testis xenografts are resistant to phthalate-induced endocrine disruption. Environ Health Perspect 2012, 120:1137-1143.
• [116]Spade DJ, Hall SJ, Saffarini CM, Huse SM, McDonnell EV, Boekelheide K: Differential response to abiraterone acetate and di-n-butyl phthalate in an androgen-sensitive human fetal testis xenograft bioassay. Toxicol Sci 2014, 138:148-160.
• [117]Gaido KW, Hensley JB, Liu D, Wallace DG, Borghoff S, Johnson KJ, Hall SJ, Boekelheide K: Fetal mouse phthalate exposure shows that Gonocyte multinucleation is not associated with decreased testicular testosterone. Toxicol Sci 2007, 97:491-503.
• [118]Liu X, He DW, Zhang DY, Lin T, Wei GH: Di(2-ethylhexyl) phthalate (DEHP) increases transforming growth factor-beta1 expression in fetal mouse genital tubercles. J Toxic Environ Health A 2008, 71:1289-1294.
• [119]Do RP, Stahlhut RW, Ponzi D, Vom Saal FS, Taylor JA: Non-monotonic dose effects of in utero exposure to di(2-ethylhexyl) phthalate (DEHP) on testicular and serum testosterone and anogenital distance in male mouse fetuses. Reprod Toxicol 2012, 34:614-621.
• [120]Song XF, Wei GH, Liu X, Zhang DY, Chen X, Deng YJ: Effects of diethylhexyl phthalate (DEHP) on INSL3 mRNA expression by Leydig cells derived from mouse embryos and in newborn mice. J INT MED RES 2008, 36:512-521.
• [121]Lehraiki A, Racine C, Krust A, Habert R, Levacher C: Phthalates impair germ cell number in the mouse fetal testis by an androgen- and estrogen-independent mechanism. Toxicol Sci 2009, 111:372-382.
• [122]McKinnell C, Mitchell RT, Walker M, Morris K, Kelnar CJ, Wallace WH, Sharpe RM: Effect of fetal or neonatal exposure to monobutyl phthalate (MBP) on testicular development and function in the marmoset. Hum Reprod 2009, 24:2244-2254.
• [123]Li LH, Donald JM, Golub MS: Review on testicular development, structure, function, and regulation in common marmoset. Birth Defects Res B Dev Reprod Toxicol 2005, 74:450-469.
• [124]Kleymenova E, Swanson C, Boekelheide K, Gaido KW: Exposure in utero to di(n-butyl) phthalate alters the vimentin cytoskeleton of fetal rat Sertoli cells and disrupts Sertoli cell-gonocyte contact. Biol Reprod 2005, 73:482-490.
• [125]Mitchell RT, Sharpe RM, Anderson RA, McKinnell C, Macpherson S, Smith LB, Wallace WH, Kelnar CJ, van den Driesche S: Diethylstilboestrol exposure does not reduce testosterone production in human fetal testis xenografts. PLoS ONE 2013, 8:e61726.
• [126]Delbes G, Levacher C, Duquenne C, Racine C, Pakarinen P, Habert R: Endogenous estrogens inhibit mouse fetal Leydig cell development via estrogen receptor alpha. Endocrinology 2005, 146:2454-2461.
• [127]Gaskell TL, Robinson LL, Groome NP, Anderson RA, Saunders PT: Differential expression of two estrogen receptor beta isoforms in the human fetal testis during the second trimester of pregnancy. J Clin Endocrinol Metab 2003, 88:424-432.
• [128]Mylchreest E, Sar M, Wallace DG, Foster PM: Fetal testosterone insufficiency and abnormal proliferation of Leydig cells and gonocytes in rats exposed to di(n-butyl) phthalate. Reprod Toxicol 2002, 16:19-28.
• [129]Ferrara D, Hallmark N, Scott H, Brown R, McKinnell C, Mahood IK, Sharpe RM: Acute and long-term effects of in utero exposure of rats to di(n-butyl) phthalate on testicular germ cell development and proliferation. Endocrinology 2006, 147:5352-5362.
• [130]Saffarini CM, Heger NE, Yamasaki H, Liu T, Hall SJ, Boekelheide K: Induction and persistence of abnormal testicular germ cells following gestational exposure to di-(n-butyl) phthalate in p53-null mice. J Androl 2012, 33:505-513.
• [131]Li H, Kim KH: Effects of mono-(2-ethylhexyl) phthalate on fetal and neonatal rat testis organ cultures. Biol Reprod 2003, 69:1964-1972.
• [132]Hutchison GR, Scott HM, Walker M, McKinnell C, Ferrara D, Mahood IK, Sharpe RM: Sertoli cell development and function in an animal model of testicular dysgenesis syndrome. Biol Reprod 2008, 78:352-360.
• [133]Mazaud-Guittot S: Dissecting the phthalate-induced Sertoli cell injury: the fragile balance of proteases and their inhibitors. Biol Reprod 2011, 85:1091-1093.