anti-Müllerian Hormone Concentrations Research Articles

Возможности коррекции гормонального статуса, репродуктивной функции и нейровегетативных расстройств у пациенток с преждевременной недостаточностью яичников

Background: nowadays reproductive health condition tends to worsen. One of the problems is a decrease in the ovarian reserve, manifested in the form of premature ovarian insufficiency (POI), which initiates the infertility problem on the one hand, and the issue of premature menopause with all its accompanying socio-psychological aspects on the other Aim: to study the therapeutic possibilities of the bovine epiphyseal cartilage polypeptide in POI and anovulation in female patients aged 30–45 years old. Patients and Methods: the study included 40 female patients aged 30–45 years with a BMI of 18.5–29.9 kg/m2 and POI signs, determined by the following indicators: follicle-stimulating hormone (FSH) concentration ≥12 IU/L; anti-müllerian hormone (AMH) 0.2–1.2 ng/mL; antral follicle count (AFC) ≤5 during ultrasound on the 7th±2 day of the menstrual cycle; decreased serum estradiol concentration and anovulation lasting 1 to 5 years or with failed IVF. The female cohort was randomized into 2 groups of 20 persons: for two menstrual cycles, the main group received a drug for menopausal hormone therapy (MHT) containing 2 mg of estradiol and 10 mg of didrogesterone (per os 1 tablet daily in a cyclic regimen from day 1 to day 28) and the bovine epiphyseal cartilage polypeptide (lyophilizate for the preparation of a solution for intramuscular administration, 10 mg intramuscularly daily for 10 days from the 5th to the 14th day of the cycle). The control group received only MHT in the same regimen for two menstrual cycles. During the study, the changes of endometrial thickness and follicle growth, ovulation markers (corpus luteum), FSH, estradiol and AMH concentrations, the number of female patients with a normalized menstrual cycle and spontaneous conception were evaluated in both groups Results: in the main group there was a statistically significant (p<0.05) decrease in FSH and luteinizing hormone (LH) levels relative to the baseline, an increase in estradiol concentration, an increase in the AFC and endometrial thickness, whereas the similar indicator changes were statistically insignificant in the control group. Besides, complete relief of neurovegetative symptoms was noted in the main group, and restoration of the ovulatory cycle was observed in 6 (30%) cases. There were no adverse events or complications during the drug regimen. Conclusion: the results obtained indicated the efficacy of the bovine epiphyseal cartilage polypeptide in the treatment of female patients with POI and anovulation and allowed to recommend the developed treatment regimen for practical use. KEYWORDS: premature ovarian insufficiency, follicle-stimulating hormone, anti-müllerian hormone, infertility, anovulation, menopausal disorders FOR CITATION: Khardikov A.V., Netyaga O.A. Correction of hormonal status, reproductive function and neurovegetative disorders in female patients with primary ovarian insufficiency. Russian Journal of Woman and Child Health. 2024;7(2):116–122 (in Russ.). DOI: 10.32364/2618- 8430-2024-7-2-5.

Assessment of anti-Müllerian hormone levels as a reproductive indicator in Japanese Black cattle.

The size of the ovarian reserve, an indicator of the number of primordial follicles, varies widely among individuals, and declines with age. However, the association between the ovarian reserve and fertility remains unclear. Therefore, in the present study, we analyzed the relationship between plasma concentrations of anti-Müllerian hormone (AMH), a marker of ovarian reserve, and reproductive outcomes in Japanese Black cattle. AMH level quartiles were positively associated with pregnancy following artificial insemination (AI), and the median number of days to pregnancy in Q4 (13 days, 95% confidence interval [CI] = 7-18 days) was significantly shorter than that in Q1 (21 days, 95% CI = 15-46 days). The odds ratio for the predicted pregnancy rate by logistic regression analysis in Q4 (4.06, 95% CI = 1.54-10.67) was also significantly higher than that in Q1. Plasma AMH concentrations were significantly higher in summer (June-August) than in winter (December-February). Furthermore, a strong correlation (r = 0.829, P < 0.001) was observed between plasma AMH concentrations at 2 and 14 months of age. Calves with plasma AMH concentrations of > 700 pg/ml at 2 months old showed a transient increase and maximum AMH concentration within 5 months of birth. Overall, the results of this study indicate that the plasma AMH concentration serves as a predictive marker for the probability of conception following AI in Japanese Black cattle. The current findings contribute to the reliable assessment of AMH production and the early prediction of reproductive performance in sexually mature heifers.

Measuring and exploring variations of serum anti-Müllerian hormone (AMH) in female fishing cat (Prionailurus viverrinus) and leopard cat (Prionailurus bengalensis)

Serum concentration of anti-Müllerian hormone (AMH) is a way to estimate the ovarian reserve in many species including domestic cats. It could therefore become an important indicator to manage breeding of wild cat species in captivity. This study focused on 1) validating the AMH Gen II ELISA test, and 2) exploring the relationship between serum AMH with age and estrus stages in fishing cats (Prionailurus viverrinus) and leopard cats (Prionailurus bengalensis). Blood samples were collected from 11 female fishing cats (5.7 ± 3.2 years old) and 8 female leopard cats (8.1 ± 1.8 years old) subjected to routine annual health check in zoo facilities. The stage of the estrus cycle was estimated from animal history, abdominal sonography and vaginal cytology. Serum AMH was measured by AMH Gen II ELISA. Intra-assay CV and inter-assay CV were 4.11 % and 5.52 %, respectively. Validation of the AMH Gen II ELISA by the linearity of sample dilution in fishing cats and leopard cats was confirmed. In fishing cats, r2 equalled 0.998 and percentage recovery was 83–101 %. In leopard cats, r2 equalled 0.999 and percentage recovery was 96–104 %. Serum AMH concentration ranged from 0.17 to 30.53 ng/mL (averaged 14.88 ± 10.03 ng/mL) in fishing cats and 1.25 to 13.52 ng/mL (averaged 6.25 ± 4.69 ng/mL) in leopard cats. There was no significant relationship between serum AMH concentration and age in both fishing cats (r = − 0.556, P = 0.075) and leopard cats (r = 0.010, P = 0.982). Likewise, there was no difference in AMH concentrations between estrus stage and other stages in both fishing cats (P = 0.069) and leopard cats (P = 0.599). In conclusion, because the limited sample size and the lack of precise information on the estrus phase, the AMH Gen II ELISA shows promising potential for detecting concentrations that may be correlated with age, estrus cycle, and ovarian reserve in future studies.

Serum anti-Müllerian hormone around the time of ovulation simulated by exogenous hormones in clouded leopards (Neofelis nebulosa).

Anti-Müllerian hormone (AMH) is produced by granulosa cells of the antral follicles. It serves as a promising biomarker for ovarian reserve and responsiveness to ovarian stimulation in humans and domestic animals. This study aimed to validate the AMH Gen II enzyme-linked immunosorbent assay (ELISA) and correlate ovarian structures with serum AMH concentrations after stimulation treatment in clouded leopards (Neofelis nebulosa). Serum samples were collected from 12 women (age 6.21 ± 3.56 years), and serum AMH concentrations were analysed using AMH Gen II ELISA. The animals were divided into two groups based on ovarian structures [preovulatory follicles (>2 mm) and/or corpora hemorrhagica] along with the presence of uterine tonicity visualized laparoscopically around the time of ovulation. Animals that exhibited these reproductive features were identified as the responder group (n = 9, aged 7.59 ± 2.96 years), whereas those lacking the corresponding features were assigned to the nonresponder group (n = 3, aged 2.06 ± 0.53 years). The intra-assay coefficient of variation (CV) and interassay CV was 3.56% and 7.75%, respectively. The linearity of AMH dilution was confirmed (r2 = .998), and the percentage of recovery ranged from 93% to 115%. The results demonstrated that overall serum AMH concentrations around the time of ovulation were negatively correlated with age (rs = -.692, p = .013). However, serum AMH concentrations were not correlated with the average number of ovarian structures (rs = -.535, p = .074). Thus, AMH Gen II ELISA was validated in clouded leopards. Around the time of ovulation, serum AMH decreased with advancing age and ovarian responsiveness cannot be evaluated using serum AMH.

Anti-Müllerian hormone: a new approach to fertility assessment in cattle – a review

The anti-Müllerian hormone (AMH) enhances fertility in cattle and hence used as a fertility marker. AMH is produced by granulosa cells of all primordial, primary, secondary follicles, and antral follicles up to 4-5 mm diameter, whereas AMH production decreases once antral stage follicles reach the 4-5 mm stage. There is an individual variation in antral follicle count (AFC) and AMH concentration. One-time determination of AMH concentration will help predict the animals’ reproductive performance at the heifer stage. In turn, it will help to cull those animals with low reproductive potential. There is no significant fluctuation in the AMH concentration during the estrous cycle, and hence it can be determined at any stage of the estrous cycle. This review explores the potential of AMH as a fertility marker and its association with the antral follicle population (AFP) as AMH can be an effective segregative tool to screen low, average or high fertility future performance in cattle.

Antimüllerian hormone level predicts ovulation in women with polycystic ovary syndrome treated with clomiphene and metformin

ObjectiveTo describe serum anti-Müllerian hormone (AMH) concentrations in a large, well-phenotyped cohort of women with polycystic ovary syndrome (PCOS) and evaluate whether AMH predicts successful ovulation induction (OI) in women treated with clomiphene and metformin. DesignSecondary analysis of randomized controlled trial Subjects333 women with anovulatory infertility attributed to PCOS who participated in thedouble blind randomized trial entitled The Pregnancy in Polycystic Ovary Syndrome I (PPCOSI) study (registration number NCT00068861) who had serum samples from baselinelaboratory testing available for further serum analysis were studied. Main Outcome MeasuresThe association between baseline AMH concentration in each of the three treatment groups and ovulation rates, pregnancy rates and live birth rates were assessed. Results322 individuals had a baseline AMH concentration available, of which the mean AMH was 11.7 ± 8.3 ng/mL with a range from 0.1 to 43.0 ng/mL. With each unit (1 ng/mL) increase in baseline AMH, the odds of ovulation decreased by 10 percent (OR 0.90, 95% CI 0.86-0.93); this effect did not differ by treatment group. Women with a high baseline AMH concentration (>8 ng/mL) were significantly less likely to ovulate compared to those with a normal baseline AMH concentration (<4 ng/mL) (OR 0.23, 95% CI 0.05-0.68). This remained statistically significant when controlling for confounders, including age, body mass index, time in study, and Homeostatic Model Assessment for Insulin Resistance (HOMA-IR) score (p=0.008). Ovulation occurred even at very high AMH concentrations; there was no maximum level noted at which no ovulation events occurred. Baseline AMH concentration was not associated with pregnancy or live birth rates when controlling for confounders (p=0.947 and p=0.848, respectively). ConclusionThese AMH values of well-phenotyped individuals with PCOS adds to the literature and will aid in identifying AMH criteria for the diagnosis of PCOS. In women with infertility and PCOS, a higher AMH concentration was associated with reduced odds of ovulation with OI with clomiphene, clomiphene+metformin and metformin.

Can AMH levels predict the need to step up FSH dose for controlled ovarian stimulation following a long GnRH agonist protocol in PCOS women?

BackgroundTo explore the role of anti-Mullerian hormone (AMH) in predicting the need to step up recombinant FSH (rFSH) dose following long GnRH agonist protocol in IVF/ICSI cycles of polycystic ovarian syndrome (PCOS) women.MethodsThis is a retrospective cohort study of 825 PCOS women undergoing long GnRH agonist protocol enrolled from Jan 2019 to Dec 2021. The daily rFSH dose at which the first response to rFSH were recorded. The dose at which the first response to rFSH was based on folliculometry during follow up in which two or more follicles reached ≥ 11 mm. A receiver operating characteristic (ROC) curve analysis was done to investigate the ability of AMH to predict the need to step up initial rFSH dose.ResultsPCOS women who needed to step up initial rFSH dose had a significantly higher AMH compared with those didn’t step up initial rFSH dose (11.37 ± 3.25ng/ml vs. 8.69 ± 3.16ng/ml, p < 0.001). In multivariate logistic regression analysis, increased AMH level was an independent factor for the need to step up initial rFSH dose in PCOS patients after adjusted for confounding factors. ROC curve analysis showed AMH could predict the need to step up initial rFSH dose (AUC = 0.738, 95%CI: 0.704–0.773), having 75.4% specificity and 63% sensitivity when the threshold AMH concentration was 9.30ng/ml. 58.8% PCOS women with AMH > 9.30 ng/ml required increased rFSH dose compared to 18.8% of women with AMH ≤ 9.30ng/ml (p < 0.001). Although the clinical pregnancy rate and live birth rate were not significantly different, there was a higher incidence of OHSS among women with AMH > 9.30 ng/ml vs. AMH ≤ 9.30ng/ml (20.8% vs. 15.3%, p = 0.043).ConclusionPCOS women with AMH > 9.30 ng/ml were resistant to rFSH stimulation and require increased dose for the cycle recruitment of ovarian follicles.

The concentration of anti-muller hormone in the blood serum of reindeer depends on the level of sex hormones

Reindeer husbandry is the leading branch of agriculture for the regions of the Far North. Reindeer (Rangifer tarandus) remains poorly studied from the point of view of genetics and reproduction, which hinders the intensification of development of the industry. Research is being actively conduct in the field of genetics of the productive qualities of reindeer, but the formation and functioning of its reproductive system has been little study. Hormonal regulation of the reproductive process and the formation of hormonal status at key stages of the reproductive cycle is a significant aspect of animal reproduction. Reindeer are characterized by a pronounced seasonality of reproduction, and the study of hormonal regulation of the reproductive cycle is the main aspect of the further use of assisted reproductive technologies. The work is devoted to the analysis of the level of anti-Mullerian hormone in females during the breeding season. The studies were carried out on reindeer farms of the MAU “Directorate of the Territory of Traditional Natural Resources Management of Soyots”, Buryatia, (female, n=29). The dependence of the concentration of anti-Müllerian hormone (AMH), which is a generally accepted indicator of the female ovarian reserve, on the concentration of sex hormones (progesterone, estradiol, testosterone and luteinizing hormone) in the blood serum of females was study. Hormone levels were determined by enzyme immunoassay. A higher content of sex hormones - testosterone, estradiol, progesterone and luteinizing hormone was revealed in the group of women with an AMH concentration of &gt;1 ng/ml than in the group of women with an AMH concentration of &lt;1 ng/ml. The significance of the differences in progesterone was p&lt;0.05, and in estradiol – p&lt;0.001. A low concentration of AMH (&lt;1 ng/ml), characterizing a reduced ovarian reserve of female reindeer, corresponds to a reduced concentration of sex hormones that support reproductive function, and, conversely, a high concentration of AMH (&gt;1 ng/ml), and, accordingly, a high ovarian reserve the reserve corresponds to an increased concentration of sex hormones - testosterone, estradiol, progesterone and luteinizing hormone.

Sildenafil as a Salvage Treatment for Ovarian Torsion: A Rat Model

OBJECTIVE: In this study, we aimed to investigate the effect of sildenafil, acting as an anti-oxidant on the ischemic rat ovarian torsion model by pointing out ovarian reserve via AMH levels which is the best marker for oocyte pool.STUDY DESIGN: This study is an experimental animal study conducted according to the principles of the Declaration of Helsinki. Twenty-seven rats were assigned into three groups, each consisting of nine individuals. Group 1: Sham operation (laparotomy only) was performed. Group 2: The torsion group had bilateral torsion and detorsion of the ovaries. Group 3: In Sildenafil and torsion group the rats received 0.7 mg/kg sildenafil (Viagra, Pfizer, New York, ABD) intraperitoneally one hour before torsion and detorsion operation. One month after the initial operation, blood was drawn via the intra-cardiac route, and serum antimullerian hormone (AMH) concentrations were studied.RESULTS: The mean weights of the rats were similar 224 g, 229 g, and 222 g for groups 1, 2, and 3 respectively. Only one rat from Group 2(T/D) died during the experiment leaving eight rats for analysis. The mean value for AMH was 24.9227±12.19124 ng/ml and the mean value for weight was 226.22±19.200. The differences in AMH groups were not statistically significant (p=0.873)CONCLUSIONS: As a result, as is shown in our study, the ovarian reserve may not be influenced by ischemia developed by ovarian torsion or any influences could not change AMH levels. Sildenafil which was given to the rats before detorsion to protect against reperfusion ischemia was found to be ineffective on ovary reserve according to AMH assessment.

Anti-Mullerian hormone profile and its association with ovarian parameters in the Chaugarkha goat

Circulating anti-Mullerian hormone (AMH) is a potential predictor of ovarian reserve in the woman and livestock. Only two such studies are reported in the goat. Accordingly, we investigated the effect of age, estrous cycle and early pregnancy on the serum AMH and its correlation with the ovarian parameters in the Chaugarkha goat. In experiment-1, a random blood sample was collected from the goats (n = 62) and sera were grouped into five based on the age distribution. The first two groups (A-B) represented pre-pubertal kids, group C represented pubertal kids while, the last two groups (D and E) indicated adult. In experiment-2, estrus was synchronized in the does (n = 30) and transrectal ultrasonography (USG) was done on alternate day till day 8 post-synchronized estrus to record antral follicle count (AFC), ovulation rate, and ovarian surface area. The does were bred in the second estrous cycle (n = 26) and pregnant ones (n = 22) were identified on day 30 post-breeding. Sera were harvested on day 0, 8, and 16 post-estrus and on gestation day 30. The concentration of AMH was assayed from the sera collected from different experiments. The results indicated that the serum AMH (ng/mL) decreased significantly from 8.37 ± 0.58–4.62 ± 0.54 during the transition from puberty to sexual maturity and remained stable in the adults (group E; 4.91 ± 0.81). Neither the estrous cycle nor early gestation significantly affected (P > 0.05) the serum AMH (ng/mL) with concentration ranging from 4.89 ± 0.50 on d8 of estrous cycle to 5.11 ± 0.56 at early gestation. Further, the serum AMH showed a significant moderate positive correlation with AFC (r = 0.61; P = 0.001), number of ovulations (r = 0.57; P = 0.002), and litter size (r = 0.59; P = 0.004). It is concluded that serum AMH has the potential to predict the AFC in the adult Chaugarkha goat.

Conservative Hypomethylation of Mesenchymal Stem Cells and Their Secretome Restored the Follicular Development in Cisplatin-Induced Premature Ovarian Failure Mice

Premature ovarian failure (POF) is one of the main causes of infertility in women under the age of 40 years. Recently, epigenetic reprogramming, particularly DNA hypomethylation, has emerged as a promising strategy to enhance the therapeutic potential of mesenchymal stem cells (MSCs). Thus, it is crucial to elucidate how far global hypomethylation of MSCs genome can maintain their pluripotency and viability and improve their therapeutic effect in chemotherapy-induced POF mice. Herein, the genomic DNA of bone marrow-derived MSCs (BM-MSCs) was hypomethylated by the DNA methyltransferase inhibitor (5-Aza-dC), and the degree of global hypomethylation was assessed by methylation-sensitive HepII/MspI restriction analysis. Next, mildly hypomethylated cells and their secretome were independently transplanted (or infused) in POF mice, established via cisplatin-mediated gonadotoxicity. We found that conservative global hypomethylation of BM-MSCs genome with low doses of 5-Aza-dC (≤0.5 μM) has maintained cell viability and MSCs-specific clusters of differentiation (CD). Engraftment of mildly hypomethylated cells in POF mice, or infusion of their secretome, improved the concentrations of estradiol (E2), follicle-stimulating hormone (FSH), and anti-Mullerian hormone (AMH). Furthermore, mice restored their normal body weight, ovarian size, and ovarian follicle count. This was associated with improved follicular development, where the populations of healthy primordial, primary, secondary, and tertiary follicles were significantly ameliorated, relative to mice transplanted with normally methylated cells. This observational study suggests that transplantation of mildly hypomethylated BM-MSCs cells and their secretome can restore the structural and functional integrity of the damaged ovaries in POF mice. Also, it presents conservative hypomethylation of BM-MSCs and their secretome as a promising alternative to MSCs transplantation.Graphical Transplantation of partially hypomethylated BM-MSCs improved the follicular count and integrity in the POF mouse model. Gonadotoxic drug (cisplatin) was used to establish the POF mouse model. In parallel, BM-MSCs were isolated, authenticated, and then incubated with the DNMTs inhibitor (5-Aza-dC). Partially hypomethylated cells and their secretome were independently transplanted into the POF mice, and both the follicular count, ovarian histology, and the serum levels of the fertility-related hormones (E2, AMH, and FSH) were assessed 1 week after transplantation or infusion. Hypomethylated BM-MSCs and their secretome increased the follicular count, increased the number of healthy follicles, and restricted apoptosis of the granulose cells. Also, the hormonal profile was improved compared to their corresponding level in mice transplanted with normally methylated cells.

An extreme model of fertility in sheep demonstrates the basis of controversies surrounding antral follicle count and circulating concentrations of anti-Müllerian hormone as predictors of fertility in ruminants

Ovarian ultrasonography and measurement of circulating concentrations of anti-Müllerian hormone (AMH) have been used to estimate follicle number and predict fertility in mammalian females, but no study has evaluated follicle number and circulating concentrations of AMH in ewes known to differ in fertility. We tested the hypothesis that ewes that had failed to lamb (BARREN) in four consecutive annual breeding seasons of 21–35 d have fewer follicles and diminished circulating concentrations of AMH compared to closely matched ewes that consistently produced lambs (FERTILE) under the same breeding protocols. Once identified, BARREN ewes (n = 19) were paired by breed and sire to a FERTILE control ewe (n = 19) and reproductive tracts were recovered at necropsy. Visible antral follicles in both ovaries were counted and a representative cross section of one ovary was embedded for histological evaluation of pre-antral follicle numbers. Paired t-tests indicated that BARREN ewes had fewer antral follicles, fewer primordial follicles, and decreased circulating concentrations of AMH compared to FERTILE ewes (P ≤ 0.01), but there were ewes in each fertility classification that had ovarian phenotypes like the opposite fertility classification. The best technologies we have currently for estimating follicle numbers are ultrasonography and measurement of circulating concentrations of AMH, but no single technique is perfect for predicting fertility. A better understanding of the under-lying biological mechanisms linking AMH, folliculogenesis, and fertility is required to improve the use of measurements of follicle number for predicting fertility in livestock.

Phenotypic and genetic relationships among anogenital distance, anti-Müllerian hormone, and in vitro embryo production in Gyr dairy cattle

Anti-Müllerian hormone (AMH) concentration and number of recovered oocytes (ROOC) are phenotypic parameters associated with in vitro embryo production (IVEP). More recently, anogenital distance (AGD) has been proposed as a proxy for fertility in dairy cattle that is easy to collect at a low cost. The aim of this study was to characterize the AGD and its phenotypic and genetic associations with AMH and IVEP in Bos indicus Gyr dairy cattle. The hypothesis was that the number of ROOC, in vitro-produced embryos, and AMH concentration would increase as the AGD decreases. From July to December 2021, a single morphometrical measurement of AGD was collected in 552 donors from 6 herds in Brazil. A subset of donors had AMH assayed on the same day. Only ovum pick-up events that occurred up to 12 mo preceding and 7 mo succeeding the AGD measurement were used to assess the association between AGD, AMH, and IVEP. Thus, 472 donors (1,551 ovum pick-up events and 140 donors with AMH) were considered in the analysis. A raw average was calculated for each individual donor's ROOC, viable oocytes, total produced embryos, viability rate, and embryo rate (defined as total produced embryos/viable oocytes). Comparisons were conducted within the age categories of 3 to <6 yr or 6 to <10 yr. Phenotypic associations were performed in SAS software (SAS Institute Inc., Cary, NC). Genetic correlations were estimated using the BLUPF90 family of programs. The AGD (128.7 mm ± 14; mean ± standard deviation) had a normal distribution and was highly variable (83 to 172 mm) among the Gyr population. Our experimental hypothesis was partially supported by a phenotypic association of a greater number of total produced embryos (R2 = 0.023) as AGD decreased. Our results failed to support an increase in AMH concentration along with a decrease in AGD. In addition, positive and low genetic correlations were observed between AGD and viable oocytes (r = 0.08), and embryo rate (r = 0.20). A greater number of viable oocytes and embryos were observed in donors in the high compared with intermediate and low ROOC categories within both age categories. The age interval of 3 to <6 yr showed a greater number of recovered and viable oocytes for the high AMH compared with the low category, but no differences were observed among the AGD categories. In summary, for the Gyr breed, AGD was phenotypically inversely associated with a quantity-related parameter, such as the total number of produced embryos. In contrast, AGD showed a low genetic correlation with qualitative-related outcomes such as viable oocytes and embryo rate. Further studies should be performed to validate these retrospective analyses and to better understand the association between AGD and IVEP.

FRI385 Influence Of Maternal Dietary Protein Precursor On Reproductive Endocrinology Of Neonatal Bovine Offspring

Abstract Disclosure: J.F. Thorson: None. L.D. Prezotto: None. Bovine oogenesis and follicular maturation are initiated during in utero development, a process that is stringently orchestrated by the endocrine milieu. Of paramount importance are the three major biologically active estrogens: estron, estradiol, and estriol that are involved in the development and maintenance of germ cells. Previous work using the current model have revealed that maternal urea (protein precursor) supplementation reduces the ovarian follicular reserve without compromising the number of secondary and tertiary follicles. Therefore, we did not anticipate an influence of dietary treatment on biologically active estrogens. However, as the ovarian reserve is positively associated with circulating concentrations of anti-Mullerian hormone (AMH) we hypothesized that offspring from dams supplemented urea would have reduced circulating concentrations of AMH at birth. To characterize the effects of maternal urea supplementation, multiparous cows pregnant with female calves were individually-fed isocaloric dietary treatments consisting of chopped forage top-dressed with a control (0 g urea/animal/d) or urea (80 g urea/animal/d) pelleted supplement. Diets met or exceeded dietary requirements throughout gestation. Blood samples were collected from offspring at birth for the collection of plasma. Plasma was analyzed for concentration of estradiol, estrone, unconjugated estriol, and AMH using commercially available ELISA kits. The effect of maternal dietary treatment on concentration of hormones in offspring were evaluated by ANOVA. As expected, maternal dietary treatment did not influence circulating concentrations of estradiol (P = 0.41), estrone (P = 0.35), or unconjugated estriol (P = 0.31). However, it was not anticipated that maternal dietary treatment would have no effect on circulating concentrations of AMH (P = 0.97) as offspring of urea-supplemented dams had fewer primordial and primary follicles than offspring from control dams. From these results we infer that maternal urea supplementation induces precocious follicular activation and/or increases the number of follicles recruited during maturation through an AMH-independent mechanism. Moreover, these data illustrate that while offspring of dams offered urea throughout pregnancy have a reduction in primordial and primary follicles, the steroidogenic competence of secondary and tertiary follicles is no different than dams offered the control diet. Presentation: Friday, June 16, 2023

Evaluation of serum anti-Müllerian hormone concentrations following treatment with vitamin D in Holstein Friesian heifers.

Anti-Müllerian hormone (AMH) levels in the blood of dairy cattle are associated with the count of antral follicles (AFC), ovarian function. Numerous studies, particularly in human medicine, have shown that vitamin D3 (Vit D3) has a positive effect on AMH levels. To our knowledge, the effects of Vit D3 on serum AMH levels in heifers have not been investigated. The goal of the current study was to evaluate the concentrations of serum AMH in dairy heifers following Vit D3 treatment. The study included 20 healthy non-pregnant Holstein Friesian heifers. These animals came to estrus at least once. All heifers received a single intramuscular dosage of 5 million IU Vit D3 (Cholecalciferol). Blood samples were collected from the coccygeal veins of all animals before and after Vit D3 injection (days 7, 14, and 28) for the measurement of AMH and 25(OH)D concentrations. In our analysis, we observed that after exogenous Vit D3 administration, circular 25(OH)D increased constantly (up to day 28 after Vit D3 injection) in all animals. On day 28, AMH concentrations saw a 10% increase in comparison with those measured the day before the Vit D3 injection. However, a comparison of AMH concentrations measured across days did not reveal statistically significant differences between Day 0, 7, 14, and 28 levels (p = .10). Furthermore, no statistical correlation was observed between the pairs (age-AMH) and (weight-AMH). Considering all times, no correlation was found between 25(OH)D and blood AMH levels. These findings demonstrated that exogenous Vit D3 did not affect serum AMH in Holstein Friesian heifers.

Polymorphisms within the &lt;em&gt;IGF1&lt;/em&gt; and &lt;em&gt;IGF1R&lt;/em&gt; genes associated with superovulation-related traits in Holstein dairy cows managed in a semiarid environment

Embryo transfer (ET) is an artificial reproductive technology used for the genetic improvement of cattle. High variation has been observed in superovulation (SPO) and embryo flush recovery, which appear to be influenced by donor cow genetics. Then, the objective was to identify single nucleotide polymorphisms (SNPs) from the genes IGF1 and IGF1R associated with reproductive traits related to SPO response in dairy cows raised in a semiarid region. Sixty-four Holstein cows were subjected to SPO, artificial insemination, and nonsurgical embryo collection. Individual blood samples were collected and used to genotype 13 SNPs from the genes IGF1 and IGF1R. Additional blood samples were collected to measure anti-Mullerian hormone (AMH) concentrations. Ovarian and embryo traits related to SPO response were evaluated. A mixed-effects model was used to identify associations between SNPs and SPO-related traits. A regression model was implemented to calculate allele substitution effects. From 13 SNPs, the SNP rs109763947 in the gene IGF1 and the SNPs rs110343126 and rs208140993 in the gene IGF1R were predictors for six traits evaluated in superovulated cows. The most favorable genotypes for these SNPs were CC, AA and GG, respectively. A linear trend was detected, suggesting an additive effect of the genes. Moreover, all traits evaluated in the current study improved as the number of favorable SNP genotypes increased, confirming a positive contribution of the SNP genes. In conclusion, three SNPs in the genes IGF1 and IGF1R were marker predictors for reproductive traits related to SPO response and embryo production in Holstein cows managed in a semiarid region.

Gut microbiota-bile acid-vitamin D axis plays an important role in determining oocyte quality and embryonic development.

To reveal whether gut microbiota and their metabolites are correlated with oocyte quality decline caused by circadian rhythm disruption, and to search possible approaches for improving oocyte quality. A mouse model exposed to continuous light was established. The oocyte quality, embryonic development, microbial metabolites and gut microbiota were analyzed. Intragastric administration of microbial metabolites was conducted to confirm the relationship between gut microbiota and oocyte quality and embryonic development. Firstly, we found that oocyte quality and embryonic development decreased in mice exposed to continuous light. Through metabolomics profiling and 16S rDNA-seq, we found that the intestinal absorption capacity of vitamin D was decreased due to significant decrease of bile acids such as lithocholic acid (LCA), which was significantly associated with increased abundance of Turicibacter. Subsequently, the concentrations of anti-Mullerian hormone (AMH) hormone in blood and melatonin in follicular fluid were reduced, which is the main reason for the decline of oocyte quality and early embryonic development, and this was rescued by injection of vitamin D3 (VD3). Secondly, melatonin rescued oocyte quality and embryonic development by increasing the concentration of lithocholic acid and reducing the concentration of oxidative stress metabolites in the intestine. Thirdly, we found six metabolites that could rescue oocyte quality and early embryonic development, among which LCA of 30mg/kg and NorDCA of 15mg/kg had the best rescue effect. These findings confirm the link between ovarian function and gut microbiota regulation by microbial metabolites and have potential value for improving ovary function.

Serum anti-Müllerian hormone concentrations are related to embryo development: lessons from time-lapse imaging.

Our objective was to study whether serum anti-Müllerian hormone (AMH) concentrations were associated with embryo morphokinetic events. This retrospective cohort study was performed in a private university-affiliated in vitro fertilization centre between March 2019 and December 2020 and included 902 oocytes cultured in a time-lapse imaging incubator, obtained from 114 intracytoplasmic sperm injection cycles performed. The relationship between AMH concentrations and morphokinetic events was investigated by considering the clustering of data (multiple embryos/patient). Evaluated kinetic markers were time to pronuclei appearance (tPNa) and fading (tPNf), time to two (t2), three (t3), four (t4), five (t5), six (t6), seven (t7), and eight cells (t8), (tSB) and time to the start of blastulation (tSB) and to blastulation (tB). Significant inverse relationships were observed between serum AMH concentrations and tPNf, t3, t4, t5, t6, t7, t8, and tB. The AMH was positively correlated with the KIDScore and implantation rate. Increased serum AMH concentrations correlated with faster embryo development. The clinical implications of this effect on embryo development warrant further investigation.

Anti-Mullerian hormone and spontaneous puberty in a diverse US Turner syndrome clinic cohort: A cross-sectional study.

Serum Anti-Mullerian Hormone (AMH) concentrations have been proposed as a marker of spontaneous puberty and future fertility in Turner syndrome (TS). Gonadotropins during minipuberty may also provide a clue to ovarian function but there is insufficient data to inform utility in the routine clinical management of TS. Our objective was to describe the distribution of AMH in a cross-sectional cohort of patients in a TS specialty clinic, and correlate with spontaneous puberty and karyotype, as well as gonadotropins during the minipuberty of infancy in a smaller subset of patients aged 2-9 months. Retrospective chart review of patients seen in the TS clinic at Children's National Hospital from 1/1/2019 to 8/24/2022, to assess AMH and correlate with karyotype and spontaneous puberty. Among 114 patients (median age 9.6 year, 0.08-22 year), AMH values were detectable in only (40/104) 38%, and higher mean AMH (2.7 ± 0.95 ng/mL) was seen in mosaic 45,X/46,XX karyotype compared to 45,X (0.03 ± 0.14 ng/mL) (p < .001), and structurally abnormal-X karyotype (0.11 ± 0.5) (p = .0003). Mean AMH was higher (1.4 ± 1.6 ng/mL) among those with spontaneous menarche compared with spontaneous thelarche but no menarche. AMH was detectable in 2/10 during minipuberty in those with the lowest luteinizing hormone (LH). Our institutional data reflects a diverse cohort of patients seen in a TS specialty clinic in the United States, showing correlation of AMH with karyotype and spontaneous menarche, as well as description of gonadotropins during minipuberty highlighting their clinical relevance. Studies in larger, prospective longitudinal cohorts will help determine their predictive value and role in the care of TS.

Can evaluation of seminal plasma and serum anti-Müllerian hormone be useful in the diagnosis of male infertility? (Literature review)

Background. Anti-Müllerian hormone (AMH) is secreted in men by Sertoli cells and plays an important role in the development of the male reproductive organs. High expression of AMH during embryonic development contributes to the regression of the Müllerian (paramesonephric) ducts; in the absence of AMH, the Müllerian ducts develop into the female internal genital organs. In adulthood, the clinical value of measuring AMH in serum and seminal plasma remains controversial. It is assumed that AMH, as a marker of the functional state of Sertoli cells, can be useful in the diagnosis of male infertility.Aim. To determine the clinical value of measuring serum and seminal plasma AMH levels in the diagnosis of male infertility by examining the current evidence base in the world literature.Materials and methods. A search was made for publications devoted to the study of the role of AMH in the diagnosis of male infertility in the international database PubMed, in the scientific electronic library eLIBRARY.RU and in the Google Scholar search engine. There were no restrictions regarding the status and period of publication.Results and conclusion. The results of the analysis indicate the absence of a significant relationship between the parameters of the ejaculate and the serum concentration of AMH, which does not allow using this indicator to assess male fertility. Serum AMH is of no value in the differential diagnosis of obstructive and non-obstructive azoospermia, the prediction of assisted reproductive technologies outcomes, and the prediction of testicular biopsy outcomes in the general non-obstructive azoospermia patient population. It is likely that serum AMH may be useful in predicting testicular biopsy results in patients with Klinefelter’s syndrome and idiopathic non-obstructive azoospermia.The clinical application of measuring the level of AMH in seminal plasma is limited by the high variability of this indicator, probably due to the dependence on the volume of the ejaculate and exposure to semen proteolytic enzymes. This indicator may be useful for the differential diagnosis of obstructive and non-obstructive azoospermia. In obstructive azoospermia, AMH is not detected in seminal plasma, but undetectable AMH concentrations can also be observed in patients with non-obstructive azoospermia, which reduces the differential value of this indicator. A promising direction is the use of AMH concentration in seminal plasma to predict the recovery of sperm motility after cryopreservation.