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We observed a modest but consistent enhancement of the activity of −3068-5′h AMH-luc after 36 h of incubation with FSH (Fig. At 2 h, the enhancement was not uniformly observed, which explains the higher variability and lack of statistically significant difference when compared with the basal condition.

Because SMAT1 cells do not express the FSH receptor (FSH-R), to test FSH action on 5′h AMH-luc activity, we cotransfected an FSH-R expression vector into SMAT1 cells and incubated them with or without 2 U/ml of recombinant human FSH.

R.), Département de Biologie, Ecole Normale Supérieure, 92120 Montrouge, France Unité de Recherches sur l’Endocrinologie du Développement (Intitut National de la Santé et de la Recherche Médicale; C. ANTI-MÜLLERIAN HORMONE (AMH), also called Müllerian-inhibiting substance (MIS), is a member of the TGF-β superfamily of growth and differentiation factors, involved in the hormonal control of sex differentiation in the male fetus (reviewed in Refs. In the testis, AMH is synthesized by Sertoli cells, the somatic component of the seminiferous tubules, from the very onset of fetal testicular differentiation (3).

We conclude that prepubertal testicular AMH production is increased by FSH stimulation through Sertoli cell proliferation and an enhancement of AMH gene transcription.

This is the first report showing the importance of distant sequences in the regulation of AMH expression.

Our results showed that FSH activates AMH transcription via adenylate cyclase, c AMP, and protein kinase A but involving a nonclassical c AMP-response pathway requiring nuclear factor-κB and activating protein 2 binding sites, which lie more than 1.9 kb upstream of the AMH transcription start site.

2), but less is known concerning the regulation of testicular AMH production after birth.

Upon binding to a specific membrane receptor in Müllerian ducts, AMH induces the regression of the anlagen of the uterus and Fallopian tubes in the male fetus (reviewed in Refs. A growing amount of evidence indicates that AMH also acts as a negative modulator of Leydig cell differentiation and testosterone secretion in the male (reviewed in Refs. The ontogeny of AMH expression in the postnatal male is characterized by the existence of high levels until puberty; then, AMH expression is significantly down-regulated (5, 6), which results in a decrease of serum AMH to very low levels (reviewed in Refs. The regulatory mechanisms involved in the activation of AMH expression in the fetal Sertoli cell have been extensively studied (reviewed in Ref. R.), Département de Biologie, Ecole Normale Supérieure, 92120 Montrouge, France Unité de Recherches sur l’Endocrinologie du Développement (Intitut National de la Santé et de la Recherche Médicale; C. in the normal fetus and neonate (6) and in the androgen insensitivity syndrome (6, 11), testicular AMH output can be stimulated by FSH (6). Using clinical and experimental approaches, we have shown that pubertal elevation of the androgen concentration within the testis is responsible for the inhibition of AMH production (reviewed in Ref. When the negative effect of androgens is absent owing either to impaired testosterone synthesis (9, 10) or to the lack of androgen receptor expression in Sertoli cells, . R.), Département de Biologie, Ecole Normale Supérieure, 92120 Montrouge, France Unité de Recherches sur l’Endocrinologie du Développement (Intitut National de la Santé et de la Recherche Médicale; C. In turn, PKA phosphorylates a class of transcription factors binding to c AMP-responsive elements (CREs) in FSH-target genes (reviewed in Ref. More recently, alternative pathways have been described for FSH signaling, involving phosphatidylinositol 3-kinase (PI3-K) and protein kinase B (PKB; Refs. In the prepubertal Sertoli cell, FSH has also been shown to increase DNA and protein synthesis resulting in cell proliferation and protein secretion (reviewed in Ref. The FSH-dependent elevation of serum AMH we have previously reported (6) could be explained by an increased AMH expression in individual Sertoli cells or by an increase in the number of Sertoli cells, or both. The best characterized pathway of FSH action in Sertoli cells is mediated through a Gα protein activates adenylate cyclase resulting in increased intracellular c AMP levels, responsible for the stimulation of protein kinase A (PKA). Furthermore, because no consensus CRE sequences have been found in the 5′-flanking sequences of the AMH gene studied to present, we investigated other potentially involved molecular pathways.

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