The orphan nuclear receptor small heterodimer partner mediates male infertility induced by diethylstilbestrol in mice
David H. Volle, Mélanie Decourteix, Erwan Garo, Judy McNeilly, Patrick Fenichel, Johan Auwerx, Alan S. McNeilly, Kristina Schoonjans, Mohamed Benahmed
David H. Volle, Mélanie Decourteix, Erwan Garo, Judy McNeilly, Patrick Fenichel, Johan Auwerx, Alan S. McNeilly, Kristina Schoonjans, Mohamed Benahmed
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Research Article Endocrinology

The orphan nuclear receptor small heterodimer partner mediates male infertility induced by diethylstilbestrol in mice

Abstract

Studies in rodents have shown that male sexual function can be disrupted by fetal or neonatal administration of compounds that alter endocrine homeostasis, such as the synthetic nonsteroidal estrogen diethylstilbestrol (DES). Although the molecular basis for this effect remains unknown, estrogen receptors likely play a critical role in mediating DES-induced infertility. Recently, we showed that the orphan nuclear receptor small heterodimer partner (Nr0b2), which is both a target gene and a transcriptional repressor of estrogen receptors, controls testicular function by regulating germ cell entry into meiosis and testosterone synthesis. We therefore hypothesized that some of the harmful effects of DES on testes could be mediated through Nr0b2. Here, we present data demonstrating that Nr0b2 deficiency protected mice against the negative effects of DES on testis development and function. During postnatal development, Nr0b2-null mice were resistant to DES-mediated inhibition of germ cell differentiation, which may be the result of interference by Nr0b2 with retinoid signals that control meiosis. Adult Nr0b2-null male mice were also protected against the effects of DES; however, we suggest that this phenomenon was due to the removal of the repressive effects of Nr0b2 on steroidogenesis. Together, these data demonstrate that Nr0b2 plays a critical role in the pathophysiological changes induced by DES in the mouse testis.

Authors

David H. Volle, Mélanie Decourteix, Erwan Garo, Judy McNeilly, Patrick Fenichel, Johan Auwerx, Alan S. McNeilly, Kristina Schoonjans, Mohamed Benahmed

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Figure 8

Nr0b2 controls G9a expression.

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Nr0b2 controls G9a expression. (A) G9a immunoblot from whole testes ...
(A) G9a immunoblot from whole testes of P10 Nr0b2+/+ and Nr0b2L–/L– untreated mice (n = 5–7 per group). Lanes were run on the same gel but were noncontiguous (white line). (B) Immunoblots of H3K9me1 or H3K9me2 performed on P10 Nr0b2+/+ and Nr0b2L–/L– mice (n = 6–7 per group). (C) mRNA levels of G9a target genes Defb42, Chst11, 110, 290, Mei1, and Akr1c13 in whole testes of P10 Nr0b2+/+ and Nr0b2L–/L– mice exposed to 0 or 0.75 μg DES (n = 10–15 per group). (A–C) Levels were normalized to actin, and normalized values of Nr0b2+/+ were set at 100%. (D) mRNA expression of Nr0b2 and G9a in F9 cells transfected with 0–800 ng pCMV-Nr0b2 (n = 3). (E) Protein accumulation of G9a in F9 cells transfected with pCMV-Nr0b2 in the absence or presence of RA (n = 3). (F and G) ChIP of crosslinked DNA from F9 wild-type or Rarg–/– cells using an anti-Rarγ antibody (F) or from F9 wild-type or Rarg–/– cells transfected with pCMV or pCMV-Nr0b2 using an anti-Flag antibody (G). Inset: A DNA sequence of ±100 bp covering either the RARE (i) or a sequence 3.0 kb upstream of the RARE (ii) was amplified. Results are expressed as fold enrichment over wild-type vehicle-treated cells (F) or over pCMV cells (G) and represent amplification variability (n = 4). *P < 0.05 versus vehicle; #P < 0.05 versus next-smallest transfected pCMV-Nr0b2 amount.