anti-Müllerian Hormone Production Research Articles

The Level of Serum Anti-Müllerian Hormone Correlates with Vitamin D Status in Men and Women But Not in Boys

Anti-Müllerian hormone (AMH) is a gonad-specific hormone, which is extensively used as a marker of gonadal status. The level of serum AMH has a high variance in similar individuals for reasons that are unknown. The AMH gene promoter contains a vitamin D response element that may cause vitamin D status to influence serum AMH levels. The objective of the study was to determine whether serum levels of AMH are related to 25-hydroxyitamin D [25(OH)D)] status. This was a correlative and intervention study. Three cohorts of participants were analyzed; mature men (n = 113), premenopausal women (n = 33), and 5- to 6-yr-old boys (n = 74). Women were given a daily supplement of ergocalciferol, cholecalciferol, or a placebo for 6 months and provided baseline and posttreatment blood samples. Serum AMH and 25(OH)D were measured and analyzed for covariation. Serum AMH positively correlated with 25(OH)D in men (r = 0.22, P = 0.02) but not boys. Both 25(OH)D and AMH levels exhibited seasonal variation in women, with an 18% decrease in AMH levels in winter compared with summer (P = 0.01). Change in AMH level correlated with the initial AMH level and the magnitude of change in vitamin D levels (r = 0.36, P = 0.004). Cholecalciferol supplementation prevented seasonal AMH change. Vitamin D may be a positive regulator of AMH production in adults, and vitamin D deficiency may confound clinical decisions based on AMH. Vitamin D deficiency should be considered when serum AMH levels are obtained for diagnosis.

Sertoli cell markers in the diagnosis of paediatric male hypogonadism

During childhood, the pituitary-testicular axis is partially dormant: testosterone secretion decreases following a drop in luteinising hormone levels; follicle-stimulating hormone (FSH) levels also go down. Conversely, Sertoli cells are most active, as revealed by the circulating levels of anti-Müllerian hormone (AMH) and inhibin B. Therefore, hypogonadism can best be evidenced, without stimulation tests, if Sertoli cell function is assessed. Serum AMH is high from fetal life until mid-puberty. Testicular AMH production increases in response to FSH and is potently inhibited by androgens. Inhibin B is high in the first years of life, then decreases partially while remaining clearly higher than in females, and increases again at puberty. Serum AMH and inhibin B are undetectable in anorchid patients. In primary or central hypogonadism affecting the whole gonad established in fetal life or childhood, all testicular markers are low. Conversely, when hypogonadism only affects Leydig cells, serum AMH and inhibin B are normal. In males of pubertal age with central hypogonadism, AMH and inhibin B are low. Treatment with FSH provokes an increase in serum levels of both Sertoli cell markers, whereas human chorionic gonadotrophin (hCG) administration increases testosterone levels. In conclusion, measurement of serum AMH and inhibin B is helpful in assessing testicular function, without need for stimulation tests, and orientates the aetiological diagnosis of paediatric male hypogonadism.

Granulosa cells are refractory to FSH action in individuals with a low antral follicle count

The reason ovarian function and fertility are diminished in women with a low antral follicle count (AFC), despite significant numbers of follicles remaining in ovaries, is unknown. The bovine model is unique to address this question because cattle and women with a low AFC exhibit similar phenotypic characteristics including a diminished ovarian reserve, reduced circulating concentrations of anti-Müllerian hormone (AMH) but heightened FSH secretion during reproductive cycles. Because women and cattle with a low AFC respond minimally to gonadotropin stimulation during IVF cycles or superovulation, granulosa cells in individuals with a low AFC are hypothesised to be refractory to FSH. The present study evaluates this hypothesis by testing whether capacity of granulosa cells to respond to FSH differs between cattle with a low and a high AFC. Granulosa cells from cattle with a low (≤15 follicles ≥3 mm in diameter) or a high (≥25 follicles) AFC were cultured with different doses of FSH. Treatments were evaluated by measurement of oestradiol (E), progesterone (P) and AMH in media and abundance of mRNAs for aromatase (CYP19A1), AMH, FSH receptor (FSHR) and oxytocin (OXT). Progesterone and OXT mRNA are well-established markers of granulosa cell luteinisation. Although high doses of FSH induced granulosa cell luteinisation, basal and FSH-induced increases in E and AMH production and expression of mRNAs for CYP19A1, FSHR and AMH in granulosa cells were much lower, while P production and OXT mRNA expression were higher in non-luteinised and luteinised granulosa cells from the low than the high AFC group. Granulosa cells in cattle with a low AFC are refractory to FSH action, which could explain why ovarian function, responsiveness to gonadotropin stimulation and fertility are diminished in individuals with a low versus a high AFC.

Oocyte yields per anti-müllerian hormone (AMH) at different female ages change with advancing female age: a new tool to assess oocyte quality?

Oocyte yields and anti-Müllerian hormone (AMH), reflecting functional ovarian reserve (FOR), decline with advancing female age. Whether they do so in parallel is unknown, and may reflect oocyte quality. Cohort study. We investigated 598 consecutive 1st IVF cycles in 62 oocyte donors and 536 infertility patients by establishing oocyte/AMH ratios, and tracking mean ratios over advancing age. Oocyte donors, after down-regulation with long agonist, were stimulated with 150-300 IU of gonadotropins. Suffering from diminished ovarian reserve, over 90% of infertility patients were stimulated with microdose agonist protocols and 450-600 IU of gonadotropins daily. The rest were down-regulated in a long luteal phase agonist protocol, and stimulated with 225-375 IU of gonadotropins. Oocyte/AMH ratios to a significant degree increase with advancing female age (R2 = 0.022; t = 3.561; P=< 0.0001), reflecting increasing oocyte production per AMH and/or decreasing AMH production per oocyte. Since AMH inhibits follicle recruitment, and attenuates FSH effects on small developing follicles, here reported findings demonstrate in a new way that the complex process of follicle maturation changes with advancing female age. These data are, thus, supportive of a new concept of ovarian aging, which no longer assumes that resting follicles/oocytes, prior to recruitment, age as women age, but, instead, that perpetually young, unrecruited oocytes enter into an aging ovarian environment for follicle maturation, which negatively affects evolving oocyte quality [Reprod Biol Endocrinol 2011;9:23]. The oocyte/AMH ratio, therefore, deserves further investigation as a promising tool in objectively assessing oocyte quality. It may also in future lend itself to assessments of treatment success with pharmaceutical agents meant to improve aging ovarian environments.

A Validated Model of Serum Anti-Müllerian Hormone from Conception to Menopause

BackgroundAnti-Müllerian hormone (AMH) is a product of growing ovarian follicles. The concentration of AMH in blood may also reflect the non-growing follicle (NGF) population, i.e. the ovarian reserve, and be of value in predicting reproductive lifespan. A full description of AMH production up to the menopause has not been previously reported.Methodology/Principal FindingsBy searching the published literature for AMH concentrations in healthy pre-menopausal females, and using our own data (combined ) we have generated and robustly validated the first model of AMH concentration from conception to menopause. This model shows that 34% of the variation in AMH is due to age alone. We have shown that AMH peaks at age 24.5 years, followed by a decline to the menopause. We have also shown that there is a neonatal peak and a potential pre-pubertal peak. Our model allows us to generate normative data at all ages.Conclusions/SignificanceThese data highlight key inflection points in ovarian follicle dynamics. This first validated model of circulating AMH in healthy females describes a transition period in early adulthood, after which AMH reflects the progressive loss of the NGF pool. The existence of a neonatal increase in gonadal activity is confirmed for females. An improved understanding of the relationship between circulating AMH and age will lead to more accurate assessment of ovarian reserve for the individual woman.

Follicle-stimulating hormone suppressed excessive production of antimullerian hormone caused by abnormally enhanced promoter activity in polycystic ovary syndrome granulosa cells

To explore: 1) the cause of excessive production of antimullerian hormone (AMH); 2) the effect of FSH on overproduction of AMH; and 3) expression of AMH receptor II (AMHRII) in polycystic ovary syndrome (PCOS) granulosa cells. Case-control study. University affiliated hospital. MATERIAL(S): The granulosa cells from 8-10-mm follicles were divided into four groups: unstimulated PCOS granulosa cells (n=12), exogenous FSH (5 ng/mL)-stimulated PCOS granulosa cells (n=12), internal FSH (150-300 IU/d)-stimulated PCOS granulosa cells (n=20), and control granulosa cells from non-PCOS infertile patients with regular menstruation (control; n=29). Exogenous FSH was added into the medium of unstimulated PCOS granulosa cells. The secretory levels and mRNA abundance of AMH, the mRNA and protein levels of AMHRII, and the luciferase activity of AMH. Both the secretiory level and mRNA abundance of AMH were significantly enhanced in PCOS granulosa cells. They could be partially suppressed by FSH. The mRNA and protein levels of AMHRII in PCOS granulosa cells were significantly increased. However, they were not affected by FSH. The luciferase activity of AMH in PCOS granulosa cells was significantly amplified but could be suppressed by FSH. Enhanced promoter activity can cause excessive production of AMH in PCOS granulosa cells. FSH may inhibit the excessive secretion of AMH by suppressing the luciferase activity of AMH promoter, but it has no effect on AMHRII expression.

Decrease of anti-Mullerian hormone in genetic spermatogenic failure

The aim of this study was to compare anti-Mullerian hormone (AMH) plasma levels in patients with azoospermia according to the physiopathology. In a prospective clinical study from April 2008 to March 2009 in University Hospital, we measured AMH levels in 49 consecutive patients with azoospermia. AMH plasma levels were correlated with FSH, inhibin B, bioavailable testosterone plasma levels and testicular volume and compared between nonobstructive azoospermia (NOA) and obstructive azoospermia (OA) and within four physiopathological subgroups of NOA: genetic, cryptorchidism, cytotoxic and unexplained. AMH, FSH, inhibin B, bioavailable testosterone plasma levels and testicular volumes were all related to each other. AMH plasma levels were lower in NOA relatively to OA. Lowest values were observed in cases of genetic NOA and on the other hand, the values observed in case of cytotoxic NOA were as high as the values observed in OA. FSH, inhibin B, bioavailable testosterone and testicular volume were not different between genetic and cytotoxic NOA. These results suggest that the decrease in AMH plasma levels is related to the origin of NOA, with low values in genetic NOA and values similar to OA in cytotoxic NOA. Further studies will be useful to understand the fine regulation of AMH production.

FSH and Its Second Messenger cAMP Stimulate the Transcription of Human Anti-Müllerian Hormone in Cultured Granulosa Cells

Anti-Müllerian hormone (AMH), also called Müllerian-inhibiting substance, a member of the TGF-ß family, is responsible for the regression of Müllerian ducts in the male fetus. In females, AMH is synthesized by granulosa cells of preantral and small antral follicles, and production wanes at later stages of follicle maturation. Using RT-PCR in luteal granulosa cells in primary culture and reporter gene techniques in the KK1 granulosa cell line, we show that FSH and cAMP enhance AMH transcription, and LH has an additive effect. Gonadotropins and cAMP act through protein kinase A and p38 MAPK signaling pathways and involve the GATA binding factor-4 and steroidogenic factor-1 transcription factors, among others. The expression profile of AMH and the dynamics of serum AMH after gonadotropin stimulation have been interpreted as a down-regulating effect of FSH upon AMH production by granulosa cells. The specific effect of gonadotropins upon granulosa cells may be obscured in vivo by the effect of FSH upon follicular maturation and by the presence of other hormones and growth factors, acting individually or in concert.

Regulation of Anti-Müllerian Hormone Production in the Cow: A Multiscale Study at Endocrine, Ovarian, Follicular, and Granulosa Cell Levels1

Anti-Müllerian hormone (AMH) is an endocrine marker that can help predict superovulatory responses to treatments administered to cows for embryo production. However, the optimal time of the estrous cycle at which a blood test should be performed for a highly reliable prognosis has not yet been established. Moreover, little is known about the regulation of AMH production. To answer these questions, a study was designed to investigate the regulation of AMH production in cows selected for their high or low ovulatory responses to superovulation. At the granulosa cell level, AMH production was inhibited by follicle-stimulating hormone but enhanced by bone morphogenetic proteins. At the follicular level, the expression of AMH within the follicle was dependent on the stage of follicular development. At the ovarian level, the size of the pool of small antral growing follicles determined ovarian AMH production. At the endocrine level, AMH followed a specific dynamic profile during the estrous cycle, which occurred independently of the follicular waves of terminal follicular development. Cows selected for their high or low responses to superovulation did not differ in the regulation of AMH production, but cows with higher responses had higher plasma AMH concentrations throughout the cycle. The optimal period of the estrous cycle at which to measure AMH concentrations with the aim of selecting the best cows for embryo production was found to be at estrus and after Day 12 of the cycle. Based on this multiscale study, we propose a model that integrates the different regulatory levels of AMH production.

Anti-Müllerian hormone in polycystic ovary syndrome and normo-ovulatory women: Correlation with clinical, hormonal and ultrasonographic parameters

Abstract Background Although the ultimate pathogenesis of polycystic ovary syndrome (PCOS) remains obscure, the distinctive feature is failure of follicular maturation resulting in anovulation and accumulation of preantral and small antral follicles which contribute significantly to the production of anti-Mullerian hormone (AMH). Objectives To compare serum AMH levels between PCOS and normo-ovulatory women; and to investigate whether AMH correlates to clinical, hormonal and ultrasonographic parameters in both groups. Design Comparative observational cross-sectional study. Setting Department of Obstetrics and Gynecology, Kasr El-Aini Teaching Hospital, Faculty of Medicine, Cairo University. Subjects Thirty-five women with PCOS according to the Rotterdam consensus; and 35 normo-ovulatory-matched controls with male, tubal or unexplained infertility. Methods Serum levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone, androstenedione, estradiol, fasting insulin and AMH were measured in the early follicular phase (day 3–4) of natural cycle or progestin-induced withdrawal bleeding (in PCOS); together with transvaginal sonography for detection of the number of small follicles ( Main outcome measures Correlation between AMH and clinical, hormonal and ultrasonographic parameters in both groups. Results AMH was significantly higher in the PCOS group. In the whole group of patients and in each group separately, AMH was positively correlated to LH, LH/FSH, number of follicles r =0.557; P =0.001 and r =0.451; P =0.007, respectively). Multiple regression analysis demonstrated that testosterone was the only determinant for AMH level ( r =0.485; P Conclusions Hyperandrogenism is associated with increased AMH secretion in PCOS patients, possibly due to increased number of small antral follicles. Assessment of AMH levels before and after the treatment of hyperandrogenism should be recommended in the plan of management of PCOS.

Anti-Müllerian hormone and polycystic ovary syndrome: a mountain too high?

Anti-Müllerian hormone (AMH) was initially thought to be produced solely by the foetal male during sexual differentiation to promote regression of the Müllerian ducts. Over the last decade, however, a new and interesting role has emerged for AMH in the ovary. In human ovaries, AMH is produced by granulosa cells from 36 weeks of gestation until menopause, with the highest expression being in small antral follicles. AMH production gradually declines as follicles grow; once follicles reach a size at which they are dominant, it has largely disappeared. Its removal from these larger follicles appears to be an important requirement for dominant follicle selection and progression to ovulation as AMH has an inhibitory role in the ovary, reducing both primordial follicle initiation and follicle sensitivity to FSH by inhibition of aromatase. It is for this reason that AMH is a focus of interest in polycystic ovary syndrome (PCOS). Serum levels are doubled, and granulosa cell production is greatly increased. Interestingly, there appear to be two groups of women with PCOS who can be distinguished by their AMH level: one group consists of those who have high levels which do not reduce with treatment and who respond less well to induction of ovulation, and a second group consists of those in whom the level is less elevated and reduces on treatment and who seem to respond rather better. Understanding the reason for the raised AMH in PCOS may give clues as to the mechanism of anovulation. To conclude, AMH appears to have a major inhibitory role during folliculogenesis, which may contribute to anovulation in PCOS.

Anti-Müllerian Hormone and Sertoli Cell Function in Paediatric Male Hypogonadism

In the prepubertal male, Sertoli cells are the most active testicular cell population. Without stimulation tests, prepubertal hypogonadism can only be evidenced if Sertoli cell function is assessed. Anti-müllerian hormone (AMH) is a distinctive marker of the prepubertal Sertoli cell. Serum AMH is high from fetal life until puberty. In postnatal life, AMH testicular production is stimulated by FSH and potently inhibited by androgens. In anorchid patients, AMH is undetectable. In prepubertal males with fetal- or childhood-onset primary or central hypogonadism affecting the whole gonad, serum AMH is low. Conversely, when hypogonadism only affects Leydig cells (i.e., LH/human chorionic gonadotrophin receptor or steroidogenic enzyme defects), serum AMH is normal/high. AMH is also normal/high in patients with androgen insensitivity. In patients of pubertal age with central hypogonadism, AMH is low for Tanner stage – reflecting lack of FSH stimulus, – but high for age – reflecting lack of testosterone inhibitory effect. FSH treatment results in serum AMH rise, whereas human chorionic gonadotrophin treatment increases testosterone levels which inhibit AMH production. In conclusion, AMH determination is helpful in assessing gonadal function, without need for stimulation tests, and orientates the aetiological diagnosis of paediatric male hypogonadism. Furthermore, serum AMH is an excellent marker of FSH and androgen action in the testis.

Short-term anti-müllerian hormone (AMH) response to human menopausal gonadotropins suggests that combined FSH + LH activities may influence AMH production by the granulosa cells (GCS)

OBJECTIVE: AMH, a by-product of the GCs of growing ovarian follicles, is likely to play key inhibitory roles during folliculogenesis. Yet, factors that regulate AMH production remain ill established and their identification could open intersting perspectives for the control of follicle growth. In normo-ovulating women, it is unclear whether gonadotropins influence AMH production. As in these patients, most of AMH-producing follicles are FSH-sensitive but devoid of LH receptors in the GCs, the effect of gonadotropins, if any, could occur directly or via steroids.

Serum anti-müllerian hormone (AMH) level/antral follicle count ratio: an alternative approach to optimize the predictability of AMH as a marker of IVF-ET outcome

OBJECTIVE: Ovarian follicles at similar developmental stages produce remarkably discrepant amounts of AMH and serum AMH levels are a function of not only follicle number but also their individual production. Considering the hypothesis that follicles producing more AMH could display increased reproductive competence, we decided to artificially extrapolate per-follicle AMH production from the ratio between serum AMH levels and small antral follicle counts (AFC) and to evaluate the predictability of this new parameter in patients undergoing IVF-ET.

Variants in the ACVR1 gene are associated with AMH levels in women with polycystic ovary syndrome

Polycystic ovaries display an increased number of pre-antral and antral follicles compared with normal ovaries, suggesting that early and late follicle development are disturbed. The pathophysiology of this process is poorly understood. Since the transforming growth factor beta family members, anti-Müllerian hormone (AMH) and bone morphogenetic proteins (BMPs), inhibit FSH sensitivity, their signalling may contribute to the aberrant follicle development in these women. Here, we investigated the role of ALK2, a type I receptor for AMH/BMP signalling, in PCOS using a genetic approach. Seven single nucleotide polymorphisms in the ACVR1 gene, encoding ALK2, were genotyped in 359 PCOS patients and 30 normo-ovulatory and 3543 population-based control women, and haplotypes were determined. Subsequently, the association of ACVR1 variants with ovarian parameters and hormone levels was investigated. The polymorphisms rs1220134, rs10497189 and rs2033962 and their corresponding haplotypes did not show different frequencies from controls, but were associated with AMH levels in PCOS women (P = 0.001, P = 0.002 and P = 0.007, respectively). Adjustment for follicle number revealed that the association with AMH levels was, in part, independent from follicle number, suggesting that variants in ACVR1 also influence AMH production per follicle. Genetic variation within ACVR1 is associated with AMH levels and follicle number in PCOS women, suggesting that ALK2 signalling contributes to the disturbed folliculogenesis in PCOS patients.

Production of anti-müllerian hormone (AMH) as a function of rhesus monkey follicle fate and growth rate during three-dimensional culture

OBJECTIVE: To evaluate Anti-Müllerian Hormone (AMH) production by preantral nonhuman primate follicle in vitro; assess whether AMH production is follicle diameter-dependent; determine the effect of follicle stimulating hormone (FSH) alone or with sequential addition of luteinizing hormone (LH) on AMH production.

Anti-Mullerian hormone is increased in follicular fluid from unstimulated ovaries in women with polycystic ovary syndrome

Anti-Mullerian hormone (AMH) may have a role in disordered folliculogenesis in polycystic ovary syndrome (PCOS). Though there have been several investigations into circulating AMH levels in patients with PCOS, no previous studies have compared AMH concentrations in the follicular fluid of unstimulated ovaries in women with PCOS with that of normally ovulating women. Follicular fluid was aspirated from 4-8-mm follicles of unstimulated ovaries during routine laparoscopy or laparotomy from women with anovulatory PCOS (n = 11) and those with regular ovulatory cycles (n = 8). Follicular AMH was compared in the two groups. Serum samples were analysed for AMH and endocrine profile. Follicular fluid AMH levels were significantly higher (P < 0.0001) in women with anovulatory PCOS (median: 466.2 ng/ml) compared with normal-ovulatory controls (median: 78.0 ng/ml). Mean follicular fluid AMH levels in PCOS patients were 60 times higher than in the serum. Moreover, there was a significant correlation between the follicular fluid and serum concentrations of AMH in the PCOS group (r = 0.86; P = 0.007) but not in controls. Highly elevated AMH in follicular fluid from PCOS patients in contrast to age-matched normal controls suggests that increased circulating concentrations of AMH are partly due to the increased production of AMH by individual follicles and not simply attributable to the increased number of small antral follicles. This suggests an intrinsic abnormality in the ovarian follicles themselves in PCOS, which could contribute to disordered folliculogenesis.

Effects of an Early Postnatal Treatment of Hypogonadotropic Hypogonadism with a Continuous Subcutaneous Infusion of Recombinant Follicle-Stimulating Hormone and Luteinizing Hormone

The neonatal-midinfancy surge in pulsatile gonadotropin secretion is attributable to an increase in GnRH pulse amplitude and is associated with a rapid expansion of Leydig and Sertoli cell populations with concomitant surges in testosterone, inhibin, and anti-Mullerian hormone production as well as an increase in testicular volume. Boys with congenital hypogonadotropic hypogonadism (HH) do not activate these processes. A potential cause for azoospermia and infertility in adult life is deficient proliferation of immature Sertoli cells before and during puberty due to the absence of FSH. The objective of the study was to investigate whether early postnatal continuous sc infusion of gonadotropins could mimic the physiological growth of testes and to evaluate responses of the Leydig and Sertoli cells to early gonadotropin replacement. Two neonates (P1 with hypotuitarism and P2 with HH) with micropenis and microorchidism were treated for 6 months with high doses of recombinant LH and FSH (a gift of Luveris and Gonal-F from Serono, Lyon, France) delivered sc with an insulin pump. Gonadotropin continuous sc infusion increased mean serum LH and FSH to normal or supranormal levels. Mean testosterone increased from undetectable levels to 7.6 and 5.2 nmol/liter, respectively, in P1 and P2. Inhibin B and anti-Müllerian hormone increased to normal levels. Mean testicular volume increased from 0.45 to 0.57 ml at birth to 2.10 ml at 7 months. Stretched penile length increased from 8 to 30 mm (P1) and 12 to 48 mm (P2). The present regimen induced physiological postnatal testes growth and high-normal activation of Leydig and Sertoli cells.

Effects of hypothyroidism on anti-mullerian hormone expression in the prepubertal rat testis.

Differentiation of adult Leydig cells (ALC) in the prepubertal rat testis is stimulated by thyroid hormone (Thy) and inhibited by the Anti-Mullerian Hormone (AMH) produced by the immature Sertoli cell (SC). As Thy induces SC maturation in the prepubertal rat testis, we hypothesized that Thy stimulation of ALC differentiation is mediated via inhibition of AMH production by the SC with their maturation. If this hypothesis is true, AMH production by the prepubertal Sertoli cells in hypothyroid rats should not decline immediately after birth as in euthyroid rats, but should be maintained throughout the hypothyroid period at a similar or higher level to that of day 1 rats. This concept was tested using control rats of postnatal days (pd) 1, 7 and 14 and hypothyroid (fed 0.1% propyl thiouracil/PTU to lactating mothers) rats of pd7 and pd14. Presence of AMH in SC was examined by immunocytochemistry for AMH. Results demonstrated that testes of pd1 rats had intense AMH positive labeling exclusively in cytoplasm of SC. In testes of pd7 and pd14 control and PTU rats, a positive but weak labeling was also observed in cytoplasm of some SC; Germ cells and testicular interstitial cells were negative for AMH at all tested ages in both experimental groups. These findings suggest that AMH production by the prepubertal SC is independent of Sertoli cell maturation and not regulated by Thy. Therefore, Thy regulation of ALC differentiation in the prepubertal rat testis is unlikely to be mediated via inhibition of AMH produced by the SC with their maturation.

Apoptosis, reactive oxygen species and follicular anti-Müllerian hormone in natural versus stimulated cycles

The aim of this study was to compare granulosa cell apoptosis, reactive oxygen species (ROS) and anti-Müllerian hormone (AMH) production in natural and stimulated cycles, and to determine their effect on IVF outcome. Of 60 women with tubal factor infertility, 30 underwent natural cycle IVF and 30 underwent ovarian stimulation with gonadotrophin-releasing hormone antagonist and gonadotrophin. Apoptosis and ROS production was measured in isolated granulosa cells from each aspirated follicle using flow cytometry. Follicular AMH concentration was measured using enzyme immunoassay. A total of 188 follicles were analysed. A lower incidence of apoptotic granulosa cells and higher mean follicular AMH concentration was observed in the natural cycle than in the ovarian stimulation group (20.2 ± 15.6% versus 35.6 ± 27.7%, P = 0.007; 6.9 ± 7.1 ng/ml versus 2.5 ± 1.7 ng/ml, P < 0.001). Ovarian stimulation increases the incidence of apoptotic granulosa cells, but has no effect on their ROS production. Ovarian stimulation reduces follicular AMH concentrations, presumably due to a negative effect of exogenous FSH on AMH production. This study was unable to explain the low pregnancy rate in natural cycle IVF using the data on apoptosis, ROS and AMH production in granulosa cells.