Single-cell sequencing reveals suppressive transcriptional programs regulated by MIS/AMH in neonatal ovaries.

Müllerian inhibiting substance (MIS/AMH), produced by granulosa cells of growing follicles, is an important regulator of folliculogenesis and follicle development. Treatment with exogenous MIS in mice suppresses follicle development and prevents ovulation. To investigate the mechanisms by which MIS inhibits follicle development, we performed single-cell RNA sequencing of whole neonatal ovaries treated with MIS at birth and analyzed at postnatal day 6, coinciding with the first wave of follicle growth. We identified distinct transcriptional signatures associated with MIS responses in the ovarian cell types. MIS treatment inhibited proliferation in granulosa, surface epithelial, and stromal cell types of the ovary and elicited a unique signature of quiescence in granulosa cells. In addition to decreasing the number of growing preantral follicles, we found that MIS treatment uncoupled the maturation of germ cells and granulosa cells. In conclusion, MIS suppressed neonatal follicle development by inhibiting proliferation, imposing a quiescent cell state, and preventing granulosa cell differentiation.

Structure of AMH bound to AMHR2 provides insight into a unique signaling pair in the TGF-ß family.

Anti-Müllerian hormone (AMH), or Müllerian-inhibiting substance, is a protein hormone that promotes Müllerian duct regression during male fetal sexual differentiation and regulation of folliculogenesis in women. AMH is a member of the transforming growth factor beta (TGF-ß) family, which has evolved to signal through its own dedicated type II receptor, AMH receptor type II (AMHR2). Structures of other TGF-ß family members have revealed how ligands infer specificity for their cognate receptors; however, it is unknown how AMH binds AMHR2 at the molecular level. Therefore, in this study, we solved the X-ray crystal structure of AMH bound to the extracellular domain of AMHR2 to a resolution of 2.6Å. The structure reveals that while AMH binds AMHR2 in a similar location to Activin and BMP ligand binding to their type II receptors, differences in both AMH and AMHR2 account for a highly specific interaction. Furthermore, using an AMH responsive cell-based luciferase assay, we show that a conformation in finger 1 of AMHR2 and a salt bridge formed by K534 on AMH and D81/E84 of AMHR2 are key to the AMH/AMHR2 interaction. Overall, our study highlights how AMH engages AMHR2 using a modified paradigm of receptor binding facilitated by modifications to the three-finger toxin fold of AMHR2. Furthermore, understanding these elements contributing to the specificity of binding will help in the design of agonists or antagonists or the selection of antibody therapies.

Markers of ovarian reserve in women living with HIV: A systematic review.

BACKGROUND: Anti-Mullerian hormone (AMH) levels indicate ovarian reserve and are predictive of reproductive aging. Studies evaluating AMH levels in women with HIV have produced conflicting results, and reasons for inter-study differences have not been assessed. To understand reproductive aging in HIV, we conducted a systematic review of ovarian reserve among women with HIV. METHODS: We searched Ovid MEDLINE, Ovid EMBASE, and CAB Direct for studies including AMH in reproductive-aged women with HIV. Two reviewers used the Newcastle-Ottawa scale to assess the quality of extracted data. RESULTS: Of the 315 reports screened, ten met the inclusion criteria. Studies were conducted across seven countries and included 3673 women with HIV and 2342 HIV-negative women in the comparison group. Ethnic distribution, combination antiretroviral therapy coverage, and viral load suppression varied considerably across studies. Nine of the ten reviewed studies reported lower unadjusted AMH levels in women with HIV than in those without HIV; however, in studies that adjusted for confounders (n = 4), only two showed an association between HIV and AMH. Low CD4 count and high viral load correlated with low AMH in the two largest studies. Other studies found that opioid use and elevated inflammatory markers were associated with low AMH. Study quality varied considerably, and many were of low quality (n = 6). CONCLUSION: Current evidence is inconclusive about the relationship between HIV and AMH, although studies suggest a trend toward lower AMH among women with HIV. Future studies that adjust for HIV-related factors, inflammatory markers, and substance use are needed in the era of contemporary HIV care to confirm the association between HIV and reduced ovarian reserve and establish its underlying cause.

CAR T Cells Targeting MISIIR for the Treatment of Ovarian Cancer and Other Gynecologic Malignancies.

The prognosis of patients diagnosed with advanced ovarian or endometrial cancer remains poor, and effective therapeutic strategies are limited. The Müllerian inhibiting substance type 2 receptor (MISIIR) is a transforming growth factor ß (TGF-ß) receptor family member, overexpressed by most ovarian and endometrial cancers while absent in most normal tissues. Restricted tissue expression, coupled with an understanding that MISIIR ligation transmits apoptotic signals to cancer cells, makes MISIIR an attractive target for tumor-directed therapeutics. However, the development of clinical MISIIR-targeted agents has been challenging. Prompted by the responses achieved in patients with blood malignancies using chimeric antigen receptor (CAR) T cell therapy, we hypothesized that MISIIR targeting may be achieved using a CAR T cell approach. Herein, we describe the development and evaluation of a CAR that targets MISIIR. T cells expressing the MISIIR-specific CAR demonstrated antigen-specific reactivity in vitro and eliminated MISIIR-overexpressing tumors in vivo. MISIIR CAR T cells also recognized a panel of human ovarian and endometrial cancer cell lines, and they lysed a battery of patient-derived tumor specimens in vitro, without mediating cytotoxicity of a panel of normal primary human cells. In conclusion, these results indicate that MISIIR targeting for the treatment of ovarian cancer and other gynecologic malignancies is achievable using CAR technology.

Follicular fluid anti-Müllerian hormone (AMH) concentrations and outcomes of in vitro fertilization cycles with fresh embryo transfer among women at a fertility center.

PURPOSE: To enhance the understanding of the clinical significance of anti-Müllerian hormone (AMH) in follicular fluid, we aimed to determine the variability of AMH concentrations in follicular fluid within and across IVF cycles and whether high follicular fluid AMH concentrations are associated with improved clinical IVF outcomes. METHODS: This was a retrospective cohort study of companion follicular fluid and serum samples from 162 women enrolled in the Environment and Reproductive Health (EARTH) Study between 2010 and 2016. AMH concentrations were quantified using a sandwich enzyme-linked immunosorbent assay. Spearman correlation and intra-class correlation (ICC) were calculated to assess variability of follicular fluid AMH, and generalized linear mixed models were used to evaluate the associations of FF AMH with IVF outcomes. RESULTS: The median (interquartile range, IQR) age of the 162 women was 34.0 years (32.0, 37.0). Follicular fluid AMH concentrations were highly correlated between follicles within each IVF cycle (Spearman r = 0.78 to 0.86) and across cycles for each woman (ICC 0.87 (95% CI 0.81 to 0.92)). Compared with women in the highest tertile of FF AMH (mean AMH = 2.3 ng/ml), women in the lowest tertile (mean AMH = 0.2 ng/ml) had lower serum AMH (T1 = 0.1 ng/ml vs. T3 = 0.6 ng/ml, p < 0.0001). In adjusted models, higher tertiles of follicular fluid AMH concentrations were associated with lower mean endometrial thickness and higher probability of clinical pregnancy. CONCLUSIONS: Follicular fluid AMH concentrations show little variability between pre-ovulatory follicles, and higher pre-ovulatory follicular fluid AMH may predict a higher probability of clinical pregnancy.

Mullerian inhibiting substance, sex hormone binding globulin and sex hormone levels in stimulant-naïve, first-diagnosed prepubertal boys with attention-deficit/hyperactivity disorder: comparison with matched healthy controls as well as before and after oros-methylpenidate treatment.

Objectives: Attention-Deficit/Hyperactivity Disorder (ADHD) is a complex neurodevelopmental disorder with strong male predominance. Since Müllerian Inhibiting Substance (MIS) produces sex-linked bias in animal studies, we aimed to investigate the role of MIS, Sex Hormone Binding Globulin (SHBG) and sex hormone levels in boys with ADHD.Methods: We compared prepubertal, psychostimulant-naïve boys with ADHD with age-matched healthy control boys (HCs). Patients were re-evaluated after 30 days of methylphenidate treatment assessing ADHD severity, and serum MIS, testosterone, estradiol, and albumin concentrations.Results: Compared to 30 HCs, with ADHD (n = 49, age = 6.9 ± 0.2 years) had lower SHBG (p = .014), and higher free testosterone (p = 0.006) and bioavailable testosterone (p = .002) percentages. Methylphenidate improved ADHD measures (all p < .0001) and abnormal baseline hormonal levels, increasing SHBG levels (p = .024), and lowering free (p = .001) and bioavailable testosterone (p = .016) percentages so that only free testosterone percentages remained higher versus HCs post-treatment (p = .02).Conclusions: Compared to age- and sex-matched HCs, prepubertal, stimulant-naïve boys with ADHD had significantly lower SHBG and higher free and bioavailable testosterone percentages, suggesting a possible contribution of sex hormones to ADHD. Osmotic-release oral system methylphenidate treatment for 30 days significantly improved ADHD symptoms and abnormal sex hormone levels, normalizing SHBG and bioavailable testosterone percentages that were similar to HCs while free testosterone remained elevated versus HCs.Key pointsCompare to healthy matched controls prepubertal stimulant-naïve boys with ADHD had significantly lower SHBG and higher free and bioavailable testosterone percentages, suggesting a possible effect on sex hormones to ADHD.After 30-day methylphenidate treatment, ADHD symptoms significantly improved, and SHBG and bioavailable testosterone percentages normalized which were similar to HCs, while free testosterone remained elevated versus HCs.We found a negative relationship between MIS levels and hyperactivity scores in ADHD boys. This finding suggests that MIS may contribute to hyperactivity symptoms, either directly by affecting behavior or indirectly by affecting sex hormone levels.

Quantification of Müllerian Inhibiting Substance/Anti-Müllerian Hormone polypeptide by isotope dilution mass spectrometry.

Measurement of serum concentrations of Müllerian inhibiting substance (MIS), also known as anti-Müllerian Hormone (AMH) by immunoassay is gaining clinical acceptance and widespread use for the diagnosis of ovarian conditions and for prediction of the response to ovarian stimulation protocols as part of assisted reproductive therapies. Provision of an International Standard to harmonize immunoassay methods is required. It is desirable for the content of a future International Standard to be assigned in mass units for consistency with the units reported by current methods. Isotope dilution mass spectrometry (IDMS), a physicochemical method with traceability to the SI (Système International d'Unités) unit of mass, is a candidate approach to provide orthogonal data to support this mass assignment. Here, we report on the development of an IDMS method for quantitation of AMH using three peptides from different regions of the AMH monomer as surrogates for the measurement of AMH. We show the sensitivity and linearity of the standard peptides and demonstrate the reproducibility and consistency of the measurement amongst the three peptides for determining the AMH content in buffered preparations and in trial preparations of recombinant AMH, lyophilised in the presence of an excess of bovine casein.

Towards international standardization of immunoassays for Müllerian inhibiting substance/anti-Müllerian hormone.

RESEARCH QUESTION: Is formulated and lyophilized, recombinant human Müllerian inhibiting substance, also known as anti-Müllerian hormone (AMH), suitable for the preparation of a WHO international standard to calibrate AMH immunoassays? DESIGN: The AMH content of a trial preparation, coded SS-581, was determined by five laboratories using seven immunoassay methods. Participants were requested to report the content of the preparation in terms of their method calibrators through the measurement of a minimum of five concentrations in the linear part of the dose-response curve. Participants were also asked to measure, concomitantly, a panel of six serum samples containing AMH at concentrations of 0.1-13.0 ng/ml. RESULTS: Across all assays, including two automated assays in development, the geometric mean content was 361.76 ng/ampoule with a geometric coefficient of variation (GCV%) of 39.95%. When measured by immunoassays that were commercially available at the time of the study, the mean content was 423.08 ng/ampoule, with a GCV% of 26.67%. The inter-method geometric means of five serum samples with an AMH concentration >0.3 ng/ml and measured concomitantly with dilutions of SS-581 varied with a range of GCV% of 14.90-22.35%, which may reflect the use of serum sample value transfer to calibrate current immunoassays, some of which use non-human AMH calibrators. The AMH in trial preparation SS-581 was shown to be biologically active in the Müllerian duct regression assay. CONCLUSIONS: A reference material prepared using human recombinant AMH is a promising candidate for the preparation of an international standard for AMH for immunoassays calibrated to recombinant human AMH.

Positive correlations of age and parity with plasma anti-Müllerian hormone concentrations in Japanese Black cows.

Anti-Müllerian hormone (AMH) is secreted from the preantral and small antral follicles. It regulates follicle development and inhibits follicular atresia. This study examined how age, parity, and time after parturition affect plasma AMH concentrations in Japanese Black cows. We measured plasma AMH concentrations in primiparous, secundiparous, and multiparous (third parity or higher) cows at four time points: day 2 (day 0 = parturition), day 8, 2 days before first postpartum ovulation (pre-1stOv), and 12 days after first ovulation (post-1stOV). We observed a positive correlation between plasma AMH concentration and age (in months) and parity on day 2, day 8, and post-1stOV, but not on pre-1stOv. The multiparous cows had higher AMH concentrations than primiparous cows throughout the postpartum period (P < 0.05). Our results indicate that age and parity significantly influence plasma AMH concentrations in Japanese Black cows during the voluntary waiting period.

Potential therapeutic applications of human anti-Müllerian hormone (AMH) analogues in reproductive medicine.

Members of the transforming growth factor-beta (TGF-beta) superfamily are key regulators of various physiological processes. Anti-Müllerian hormone (AMH) which is also commonly known as Müllerian-inhibiting substance (MIS) is a member of the TGF-beta superfamily and an important regulator of reproductive organ differentiation and ovarian follicular development. While AMH has been used for diagnostic purposes as a biomarker for over 15 years, new potential therapeutic applications of recombinant human AMH analogues are now emerging as pharmacologic agents in reproductive medicine. Therapeutic uses of AMH in gonadal tissue may provide a unique opportunity to address a broad range of reproductive themes, like contraception, ovulation induction, onset of menopause, and fertility preservation, as well as specific disease conditions, such as polycystic ovarian syndrome (PCOS) and cancers of the reproductive tract. This review explores the most promising therapeutic applications for a novel class of drugs known as AMH analogues with agonist and antagonist functions.

Sex-dependent expression of anti-Müllerian hormone (amh) and amh receptor 2 during sex organ differentiation and characterization of the Müllerian duct development in Xenopus tropicalis.

Amphibian gonadal differentiation involves the action of sex steroids. Recent research indicates that the anti-Müllerian hormone (AMH) is involved in testicular development in some lower vertebrate species. For amphibians there is a lack of data on ontogenetic expression of the AMH receptor AMHR2/amhr2 and of progesterone receptors (PGRS/pgrs). Here we expand the knowledge on amphibian sex differentiation by characterizing ontogenetic mRNA levels of amh, amhr2, intracellular and membrane pgrs (ipgr and mpgr beta) and cytochrome P450 19a1 (cyp19a1) (ovarian marker) in the urogenital complex of the model species Xenopus (Silurana) tropicalis. Furthermore, we characterized the ontogenetic development of the Müllerian ducts (precursors of the female reproductive tract) histologically. The developmental period investigated spanned from beginning of gonadal differentiation, Nieuwkoop and Faber (NF) stage 51, to 4weeks post-metamorphosis. The Müllerian ducts were first observed at NF 64 in both sexes. Male-enhanced amh mRNA levels from NF 53/54 to 6days post-metamorphosis and female-enhanced cyp19a1 levels from NF 53 to 4weeks post-metamorphosis were noted. The sexually dimorphic mRNA level profile was more distinct for amh than for cyp19a1. The pgrs mRNA levels increased over the studied period and showed no sex differences. At later developmental stages, the amhr2 mRNA level was increased in putative females compared with males. Our findings suggest that AMH has a role in gonadal differentiation in X. tropicalis. We propose relative gonadal amh mRNA level as a testicular marker during early gonadal development in amphibians.

Impact of metformin on anti-Müllerian hormone in women with polycystic ovary syndrome: a secondary analysis of a randomized controlled trial.

Conclusions on the effect of metformin on circulating anti-Müllerian hormone (AMH) levels in women with polycystic ovary syndrome (PCOS) are ambiguous. We performed a secondary analysis of a randomized, double-blind, placebo-controlled cross-over trial. Fifty-six women with hyperandrogenemic PCOS were included. Each woman served as her own control receiving a daily dose of either 1700 mg metformin or placebo for 6 months. After a 3-month wash-out period they received the opposite treatment. The decrease in AMH from a median of 49.5 to 46.9 pmol/L after 6 months on metformin was overall not significant (p = 0.81), nor were changes in obese women (from 49.5 to 38.2 pmol/L; p = 0.53). Comparing individual metformin/placebo AMH values, a small absolute decrease of 9.3 pmol/L (p = 0.03) was observed in obese women after 6 months relative to baseline, suggesting a trend towards decreasing values after metformin treatment, mainly in obese women.

AMH concentration is not related to effective time to pregnancy in women who conceive naturally.

This study determined whether anti-Müllerian hormone (AMH) concentration influences the time necessary to conceive a live-born child--effective time to pregnancy (eTTP)--in a population of women who conceived naturally. This is an observational study of 87 women with a planned spontaneous pregnancy resulting in a live birth. eTTP was assessed retrospectively by a questionnaire and AMH was measured in a frozen serum sample from first trimester of pregnancy. eTTP was correlated with age (r=-0.24, P=0.02), but not with AMH (r=-0.10) or body mass index (r=0.05). With logistic regressions, the only variable that affected the probability of pregnancy within 3 or 6 months was age, irrespective of whether an AMH concentration limit of 1.0 ng/ml or 2.0 ng/ml was chosen. In conclusion, this study suggests that there is no relationship between AMH concentration and eTTP and therefore speaks against determining AMH in women who are not infertile for the purpose of predicting their chances of pregnancy. The findings are concordant with previous reports describing AMH as a quantitative but not a qualitative marker of ovarian reserve and therefore does not reflect a woman's ability to become pregnant. Anti-Müllerian hormone (AMH) is secreted by small growing ovarian follicles and reflects a woman's ovarian reserve - the number of primordial follicles at a given time. AMH concentrations has been extensively studied in infertile women but there are only scarce data on AMH in non-infertile women. Our objective was to determine whether AMH concentrations influence the time necessary to conceive a live-born child - also called effective time to pregnancy (eTTP) - in a population of women who conceived naturally. We conducted an observational study between 2007 and 2009 in which we assessed eTTP retrospectively in 87 women who had delivered a live-born child and measured AMH in a frozen blood sample collected during the first trimester of pregnancy. The results of our study show, as expected, a decrease of AMH concentrations as age increases but no relationship between AMH and eTTP. In conclusion, our study results suggest AMH concentrations do not influence the time necessary to conceive a live-born child spontaneously and therefore speak against determining AMH in women who are not infertile for the purpose of predicting their chances of pregnancy. Our findings are concordant with previous reports describing AMH as a quantitative but not a qualitative marker of ovarian function that does therefore not reflect a woman's ability to become pregnant.

Comparing Serum AMH, InhB, Testosterone Levels and Finger Length Ratio (2D/4D) of Male Children with Specific Learning Disorder and Controls.

Introduction: It has been suggested that inhibin B (InhB), Anti-Müllerian hormone (Müllerian-inhibiting substance, AMH) levels, and 2D/4D finger length ratios are related to sex differences in neurodevelopmental disorders. The aim of this study is to investigate the role of InhB, AMH levels, and 2D/4D finger length ratios in male children with specific learning disorder (SLD). Methods: The study included 38 male children diagnosed with SLD and 38 males of similar ages without SLD as the control group. Tests used in the evaluation were the Kiddie Schedule for Affective Disorders and Schizophrenia for School-Age Children-Present and Lifetime version, Specific learning disorder clinical observation battery, Wechsler Intelligence Scale for Children-Revised (WISC-R), and Conners' Parent Rating Scale. Revised: Short Form. Serum AMH, InhB, and Testosterone levels were measured using an enzyme-linked immunosorbent assay. Results: Male children diagnosed with SLD demonstrated significantly higher levels of serum InhB compared to controls (t= 2.59 p=0.009); both groups had similar levels of serum testosterone and AMH. The 2D/4D finger ratios in the SLD group were found to be lower than those in the control group (t= 2.92 p= 0.005). Serum InhB levels were positively correlated with WISC-R verbal scores (p= 0.003). Conclusion: Our findings suggest that serum InhB levels and the 2D/4D ratio, which is an indicator of prenatal testosterone exposure, may play a role in the male predominance of SLD.

Comparison of serum anti-Müllerian hormone between unilateral and bilateral ovarian endometriomas during follicular, luteal, and random menstrual phases: a retrospective study.

BACKGROUND: Over the last two decades, serum levels of anti-Müllerian hormone (AMH) have been shown to be reliable markers of ovarian reserve. This study aimed to compare baseline serum AMH levels and well-controlled clinical factors between patients with unilateral and bilateral ovarian endometriomas during the menstrual phase. METHODS: We conducted a retrospective study. We enrolled 136 patients aged 18 to 36 years who were diagnosed with unilateral or bilateral ovarian endometriomas. Serum AMH levels of all patients and their latest two to three menstrual cycles were measured before surgery for ovarian endometriomas. The latest menstrual cycle length ranged from 26 to 30 days. Patients with irregular menstruation, a recent medication history of hormonal drugs other than oral contraceptive pills, a previous history of ovarian surgery, or any medical history influencing ovarian function were excluded. RESULTS: Of the 136 patients, 76 (55.9%) had unilateral ovarian endometriomas and 60 (44.1%) had bilateral ovarian endometriomas. Serum AMH levels were not significantly different between the two groups in the follicular phase, luteal phase, or at any random time point. CONCLUSION: Serum AMH levels were not significantly different between unilateral and bilateral ovarian endometriomas in the follicular and luteal phases, or at any random time during the menstrual cycle when various confounding factors were excluded.

Breast cancer risk coordinators: Artificial intelligence-based density measurement and Mullerian-inhibiting substance.

BACKGROUND: Due to its increasing prevalence, breast cancer has become a serious public health problem. In addition to the models used to identify individuals at risk, the search for fast and accurate tools has continued for years. AIMS: In our study, we aimed to examine the correlation of mammographic density measurement and serum Mullerian-inhibiting substance (MIS) levels with an effective model such as Gail. METHODS: Of the women whose serum MIS levels were measured in the last 1 year, 214 participants who applied for routine breast examination were included in the study. The age range was between 40 and 60. Exclusion criteria were determined as pathological mammographic findings, active breast symptom, and thoracic radiotherapy history. Mammographic density measurement (PD) was performed with the artificial intelligence-based Deep-LIBRA software. The relationship of these two parameters with the lifetime risk of developing breast cancer was examined. RESULTS: The correlation between PD and GRP was remarkable (p < 0.01 cc:0.35). A positive correlation was observed between serum MIS levels and increased breast cancer, but it was not possible to prove this statistically (p = 0.056). It was thought that this situation was caused by perimenopausal patients. Because when the menopause group was excluded, the correlation between MIS levels and GRP decreased (p = 0.12 cc:0.17). CONCLUSIONS: PD measurement can be considered as a promising method for the determination of individuals at risk for breast cancer in a large group of patients, but we think that serum MIS levels are not suitable for risk assessment in perimenopausal patients.

Anti-Müllerian hormone in queens: Serum concentrations and total ovarian follicle population.

The objectives of this study were: a) To report anti-Müllerian hormone (AMH) serum concentrations in neonatal, pre and postpubertal female cats. b) To establish the relationship between serum AMH with either age and estrous cycle c) To correlate the total number of different ovarian follicle types with AMH in adult queens. A single blood sample was collected from 10 neonates (including 5 male), 15 prepubertal and 48 postpubertal female cats to measure AMH. Eight, 10, and 18 of this latter group were in follicular (FP), luteal phase (LP), and anestrus (AN), respectively. The total number of each follicle type was histologically counted using the Gougeon and Chainy (1987) formula in a subgroup of 10 adult queens. Overall AMH mean of these the female cats was 6.31 ± 0.54 ng/mL. The neonatal females had lower AMH serum concentrations than their male littermates (2.56 ± 0.49 vs. >23 ng/mL; P < 0.01). Concentrations were also higher in prepubertal than in neonatal and postpubertal cats (11.79 ± 1.36 vs. 2.56 ± 0.49 vs. 4.87 ± 0.38 ng/mL; P < 0.01). Queens below 12 mo of age had the highest AMH levels (10.41 ± 1.16; P < 0.01). Age was inversely correlated with AMH (r = -0.5; P < 0.01). Animals in FP had lower AMH concentrations than AN females (2.51 ± 0.33 vs. 5.46 ± 0.76 ng/mL; P < 0.05). No difference in the total number of each follicle type were found between either ovary (P > 0.05). A high correlation was only found between small antral follicles and AMH concentrations (r = 0.85; P < 0.01). It was concluded, that AMH can provide an indirect, reliable marker for the assessment of ovarian follicle size and functionality. Age as well as pubertal state should be considered when evaluating AMH concentrations in this species.

Persistent Mullerian duct syndrome with polycystic ovary in a young adult: A rare case report.

Persistent Mullerian Duct Syndrome (PMDS) is a type of pseudohermaphroditism that occurs in males. It is an autosomal recessive type of familial disease that is commonly associated with a history of consanguinity. We have documented this case of a 22-year-old adult male who came with acute right iliac pain; after an ultrasound scan and hormone investigations, he was diagnosed with polycystic ovarian syndrome (PCOS).

Y-specific amh allele, amhy, is the master sex-determining gene in Japanese flounder Paralichthys olivaceus.

Japanese flounder (Paralichthys olivaceus) is an important marine fish species of both fisheries and aquaculture in Northeast Asia. The commercial interest for all-female progenies due to several sex-related traits has prompted basic research on the mechanisms of sex determination in this species. By conducting a linkage analysis of the sex-determining locus, we initially identified 12 microsatellite markers linked to sex in 11 scaffolds, whose localization was restricted to a specific region of linkage group 9. Sequence analysis of this region identified 181 genes based on the UniProt database annotations. Among them, the amh gene was considered a potential candidate for sex determination because this gene is known to have taken over the role of sex determination in many teleosts. An in-depth sequence analysis of both the coding and non-coding regions of amh in XX and XY individuals detected nine SNPs linked with maleness. However, because these substitutions were synonymous, the upstream and downstream regions of amh were also investigated and a male-specific variant with deletions in the promoter region was detected. This truncated Y-specific amh variant was named amhy, and the amh shared by both sexes was named amhx. The association analysis using both females and males of the genotypic sex inferred by the presence/absence of amhy found complete association with phenotypic sex and genotype. Gene expression analysis in larvae derived from a single-pair progeny by quantitative real-time PCR detected amhy transcripts in the larval trunks between 20 and 100 days after hatching only in XY larvae. Localization of amhy by in situ hybridization was detected in presumptive Sertoli cells of XY gonads. Expression of amhx was almost undetectable in both XX and XY genotypes. Loss of Amh function by CRISPR-Cas9 induced male-to-female sex reversal, indicating that this gene was necessary for the masculinization of XY individuals. In conclusion, the complete linkage of amhy with males, its early expression in XY gonads before testicular differentiation, and the induction of sex reversal by loss-of-function mutation support the view that amhy is the sex-determining gene in this species.

Anti-Müllerian Hormone and Polycystic Ovary Syndrome in Women and Its Male Equivalent.

This article reviews the main findings on anti-Müllerian hormone (AMH) and its involvement in the pathogenesis of polycystic ovary syndrome (PCOS) and its male equivalent. In women, AMH is produced by granulosa cells from the mid-fetal life to menopause and is a reliable indirect marker of ovarian reserve. AMH protects follicles from atresia, inhibits their differentiation in the ovary, and stimulates gonadotrophin-releasing hormone neurons pulsatility. AMH overexpression in women with PCOS likely contributes to the increase of the follicle cohort and of androgen levels, leading to follicular arrest and anovulation. In the male, AMH is synthesized at high levels by Sertoli cells from fetal life to puberty when serum AMH falls to levels similar to those observed in women. AMH is involved in the differentiation of the genital tract during fetal life and plays a role in Sertoli and Leydig cells differentiation and function. Serum AMH is used to assess Sertoli cell function in children with disorders of sex development and various conditions affecting the hypothalamic-pituitary-testicular axis. Although the reproductive function of male relative of women with PCOS has been poorly investigated, adolescents have elevated levels of AMH which could play a detrimental role on their fertility.