Six Decades of Research on Human Fetal Gonadal Steroids

Acknowledgments: All authors wish to dedicate this review to Bernard Jégou who enabled their meeting, and who was a perpetual source of inspiration for their research on human fetal gonads and in particular on their steroidogenic capacities with testes as ovaries. Funding: This work was funded by the European Union’s Horizon 2020 FREIA (grant agreement No. 825100) and the Marie Skłodowska‐Curie PROTECTED projects (grant agreement No. 722634 to P.A.F.). P.A.F., S.C.P., P.L., and L.L. are FREIA project recipients. T.L. was a recipient of funding from the French agency for food and safety (Anses).Peer reviewedPublisher PD

Ibuprofen alters human testicular physiology to produce a state of compensated hypogonadism

correction de l'article correspondant à la notice https://prodinra.inra.fr/record/425677International audienceConcern has been raised over increased male reproductive disorders in the Western world, and the disruption of male endocrinology has been suggested to play a central role. Several studies have shown that mild analgesics exposure during fetal life is associated with antiandrogenic effects and congenital malformations, but the effects on the adult man remain largely unknown. Through a clinical trial with young men exposed to ibuprofen, we show that the analgesic resulted in the clinical condition named "compensated hypogonadism," a condition prevalent among elderly men and associated with reproductive and physical disorders. In the men, luteinizing hormone (LH) and ibuprofen plasma levels were positively correlated, and the testosterone/LH ratio decreased. Using adult testis explants exposed or not exposed to ibuprofen, we demonstrate that the endocrine capabilities from testicular Leydig and Sertoli cells, including testosterone production, were suppressed through transcriptional repression. This effect was also observed in a human steroidogenic cell line. Our data demonstrate that ibuprofen alters the endocrine system via selective transcriptional repression in the human testes, thereby inducing compensated hypogonadism

Parallel assessment of the effects of bisphenol A and several of its analogs on the adult human testis

International audienceStudy question - Are bisphenol A (BPA) and BPA analogs (BPA-A) safe for male human reproductive function? Summary answer - The endocrine function of human testes explants [assessed by measuring testosterone and insulin-like factor 3 (INSL3)] was impacted by exposure of the human adult testis explants to BPA/BPA-A. What is known already - The few epidemiologic studies performed suggest that bisphenols have potential endocrine disruptive properties, but they did not identify clear and direct patterns of endocrine disruption. Study design, size, duration - Adult human testis explants in culture were exposed to BPA and the analogs bisphenol F (BPF), bisphenol S (BPS), bisphenol E (BPE), bisphenol B (BPB) and bisphenol A diglycidyl ether (BADGE) at 10-9-10-5 M for 24 or 48 h. Participants/materials, setting, methods - Human adult testes were obtained from prostate cancer patients who had no hormone therapy, or from multiorgan donors. After ex vivo exposure to the investigated bisphenols, the measured outcomes were related to histopathology (gross morphology and germ cell viability determined by anti-caspase three immunohistochemistry), and the levels of testosterone, INSL3 and inhibin B were measured using immunoassays. The levels of mRNA encoding key enzymes of bisphenol biotransformation were investigated by quantitative PCR: UGT2B15 UDP (glucuronosyltransferase two family, polypeptide B15), GUSB (glucuronidase beta), SULT1A1 and 3 (sulfotransferase family 1 A member 1 and 3) and STS (steroid sulfatase). Main results and the role of chance - A significant dose-dependent inhibition was found between testosterone levels measured in the culture medium and concentrations of BPA (P = 0.00778 at 24 h and P = 0.0291 at 48 h), BPE (P = 0.039) and BPF (P = 0.00663). The observed BPA and BPA-A-induced inhibition of testosterone production varied according to duration of exposure and BPA/BPA-A concentrations. BPA (10-9 M; P < 0.05), BPB (10-9 M; P < 0.05), BPS (10-9 and 10-8 M; P < 0.05) and BADGE (10-5 M; P < 0.05) increased Leydig cell INSL3 production. By contrast, BPE dose dependently inhibited INSL3 (P = 0.0372). Conversely, Sertoli cell function (inhibin B) and germ cell viability were not significantly affected by either bisphenols. Large scale data - N/A. Limitations, reasons for caution - Environmental compounds cannot be deliberately administered to men, justifying the use of an ex vivo approach. A relatively low number of testes samples were available for analysis (n = 3, except for testosterone secretion with n = 5). The active concentrations of BPA and BPA-A used in the study were higher than those found in human biological fluids. Wider implications of the findings - Under our experimental conditions, direct exposure to BPA or BPA-A can result in endocrine disturbance in the adult human testis. Study funding/competing interest(s) - This study was funded by Inserm (Institut National de la Santé et de la Recherche Médicale), EHESP-School of Public Health, University of Rennes1, by grants from the Agence Nationale de la Recherche (ANR; grant#ANR-13-CESA-0012-03 NEWPLAST) and Agence Nationale de Sécurité Sanitaire de l'Alimentation, de l'Environnement et du Travail (ANSES; grant#EST-2010/2/046 (BPATESTIS)). All authors declare they have no current or potential competing financial interests

Antiepileptic drugs are endocrine disruptors for the human fetal testis ex vivo

International audienceValproic acid (VPA) has long been the most widely used antiepileptic drug (AED) for the treatment of epilepsy, bipolar psychiatric disorders, and migraine. However, long-term VPA treatment has several adverse effects on the male reproductive system notably on endocrine functions and/or spermatic parameters. In utero exposure of the fetus to VPA is well known to be associated with a higher risk of several congenital malformations including those of male reproductive organs. Subsequent generations of AEDs, such as carbamazepine (CARB) and lamotrigine (LAM), are considered safer and are currently recommended for women of child-bearing age with epilepsy. Because anomalies of the male genital tract mostly result from endocrine imbalance during fetal life, we hypothesized that AEDs could directly impair testis differentiation. We thus aimed at identifying and characterizing the effects of VPA, CARB, and LAM on the differentiation and function of the different testicular cell types, and at understanding the mechanisms underlying these effects. By using ex vivo culture of first-trimester human fetal testes, we show that VPA induces multiple endocrine disruptive effects, compared with the milder ones caused by CARB and LAM. AED also subtly altered the germ cell lineage in distinct manners. Transcriptomic analysis of VPA-induced alterations highlighted a very broad range of effects on the fetal testis. Overall, our results show that AEDs can behave as endocrine disruptors for the human fetal testis ex vivo. This is consistent with, and likely underlies, the VPA-induced male genital tract masculinization abnormalities observed in patients

Paracetamol, aspirin and indomethacin induce endocrine disturbances in the human fetal testis capable of interfering with testicular descent.

International audienceContext: Masculinization depends on the fetal testis. Exposure of the human fetus during pregnancy to paracetamol or/and to other mild-analgesics is associated with an increased risk of cryptorchidism. Objective:We aimed at determining whether mild analgesics disrupted the morphology and endocrine function of the human testis.Design:We used an in vitro system based on the culture of human fetal testes exposed or not to paracetamol, its metabolite AM404, aspirin, indomethacin, ketoconazole at 10(-4) M to 10(-7) M.Setting:The study was conducted at the University of Rennes I.Patients/Participants:Human fetal testes were from pregnant women following induced abortion, between 7 and 12 weeks of gestation (GW). Main Outcome Measures:Testosterone (radioimmunoassay: RIA), Anti-Müllerian Hormone (AMH; ELISA), insulin-like factor 3 (INSL3; RIA), prostaglandin D2 and E2 (PGD2 and PGE2, respectively; ELISA), were assayed in the media. Testicular cells were counted using histology and image analysis. The possible nuclear receptor-mediated activities of the analgesics were investigated using reporter cell lines expressing estrogen (ERs), androgen (AR) and peroxysome proliferator-activated γ ( PPARγ) receptors.Results:Indomethacin and aspirin stimulated testosterone production, particularly by the younger testes (8-9 GW vs 10-12 GW). Paracetamol, AM404 and ketoconazole decreased INSL3 levels. Aspirin stimulated, whereas ketoconazole inhibited AMH production. PGE2 levels were inhibited by paracetamol, and aspirin in the 7-12 GW testes, and by indomethacin but only in 7-9.86 GW old testes. The inhibitory trends seen for PGD2 were not statistically significant. Conclusions:Analgesics at concentrations relevant to human exposure cause endocrine disturbances in the fetal testis. We suggest that the fetal human testis displays slight critical age windows for sensitivity to direct exposure to aspirin, indomethacin and paracetamol. The analgesic-induced inhibition of INSL3 may be the mechanism by which analgesics increase the risk of cryptorchidism. Greater caution is required concerning consumption of analgesics during pregnancy

An investigation of the endocrine-disruptive effects of bisphenol a in human and rat fetal testes.

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Representative immunostaining of CYP11A1 of human fetal testis explants 11–12 GW.

<p>The 3,3-diaminobenzidine tetrahydrochloride staining appears dark brown in all panels, and sections were counterstained with hematoxylin. Testis cords could be easily delineated in all treated explants except in the sections of explants that have been exposed to BPA 10^-5M without gonadotrophin, in which the boundaries of the testicular cords appeared somewhat blunted. Scale bar corresponds to 100 μm.</p

INSL3 production after culture of 8–12 GW human fetal testis explants in absence (Ctrl) or presence of 10^-8M-10^-5M.

<p>A) INSL3 production after culture of human fetal testis explants in the presence of BPA and hCG, hLH or no gonadotrophin (-gonado) supplementing the medium (RIP). INSL3 concentrations after culture of 8–12 GW human fetal testis in the presence of ethanol (Ctrl) or from 10^-8–10^-5 M BPA. Results are expressed as normalized production of INSL3 of treated samples as the percentage of that of the respective untreated first day of culture sample (RIP). n = 3–9 for hLH group, n = 3–11 testes for hCG group and n = 6–9 for the group without gonadotrophin. Values are means +/- SEM. *p<0.05, **p<0.01, ***p<0.001 (two-way ANOVA, followed by a Holm-Sidak test or a Wilcoxon test to compare matched sample). B) INSL3 production represented as normalized production of INSL3 of treated samples as the percentage of that of the respective untreated first day of culture sample and control (RIP, %Ctrl). INSL3 concentrations after culture of 8–12 GW human fetal testis explants in the presence of ethanol (Ctrl) or from 10^-8–10^-5 M BPA. Results are expressed as normalized production of INSL3 of treated samples as the percentage of that of the respective untreated first day of culture sample and control (RIP, %Ctrl). n = 3–9 for hLH group, n = 3–11 for hCG group and n = 6–9 for the group without gonadotrophin. Values are expressed as least squares means +/- SE. *p<0.05, **p<0.01, ***p<0.001 (two-way ANOVA, followed by a Holm-Sidak test or a Wilcoxon test to compare matched sample).</p

Effects of BPA on testosterone production after 72h of culture of 7–12 GW human fetal testis explants.

<p>A) Dose-dependent effects of BPA—diluted in DMSO or ethanol—with hLH on testosterone production by human fetal testis explants (RTP; RTP, %Ctrl): BPA was diluted in DMSO or ethanol and the media collected after 72 hr of culture were assayed. Results are expressed as normalized production of testosterone of treated samples as the percentage of that of the respective untreated first day of culture sample (RTP) (top) and as the percentage of that of the respective untreated first day of culture sample and control (RTP, %Ctrl) (bottom). Values are mean +/- SEM of testosterone from the respective untreated first day of culture basal sample and control. The number of testes (n) is indicated below the graphic for each condition. Dose-responses were analyzed for significance with a Wilcoxon test. The effects of the vehicle were analyzed with two-way ANOVA. *p<0.05. B) Testosterone production after culture of human fetal testis in the presence of BPA and hCG, hLH or no gonadotrophin (-gonado) supplementing the medium (RTP). n = 7–13 testes for the hCG group, n = 3–20 testes for hLH group and n = 9–21 in the absence of gonadotrophin. *p<0.05, ***p<0.001 (two-way ANOVA followed by a Holm-Sidak test or a Wilcoxon test to compare matched samples). C) Testosterone production represented as a fold change from the respective first day of culture sample and control (RTP, %Ctrl). Results are expressed as normalized production of testosterone of treated samples as the percentage of that of the respective untreated first day of culture sample (RTP) and control (RTP, %Ctrl). n = 7–13 testes for the hCG group, n = 3–20 testes for hLH group and n = 9–21 in the absence of gonadotrophin. *p<0.05, ***p<0.001 (two-way ANOVA followed by a Holm-Sidak test or a Wilcoxon test to compare matched samples).</p