PSX-1 Effects of maternal stressors on testicular development in utero in beef cattle.

Anti-Müllerian hormone profiling in prepubertal horses and its relationship with gonadal function

A Sertoli Cell-Specific Knockout of Connexin43 Prevents Initiation of Spermatogenesis

Effects of azithromycin exposure during pregnancy at different stages, doses and courses on testicular development in fetal mice

Changes in peripheral anti-Müllerian hormone concentration and their relationship with testicular structure in beef bull calves

Anti-Müllerian hormone (AMH), known for its role during foetal sexual differentiation, is secreted by the Sertoli cells in males and the granulosa cells in females during post-natal life. As serum AMH concentrations correlate with follicle numbers, AMH is utilized as a marker of ovarian reserve in many species. In dogs and cats, AMH is used as a diagnostic tool to determine spay or neuter status. In the available literature, no research regarding serum AMH levels in rabbits has been published yet. The objectives of the present study were to (1) measure serum AMH concentrations in female rabbits and investigate the value of AMH as a diagnostic tool to differentiate between spayed and intact does and (2) relate measured AMH levels to pseudopregnancy and ovarian follicle numbers. For AMH measurement, serum samples were obtained from sexually intact (n=64) and spayed (n=22) female rabbits. Spayed does were of various breeds; intact rabbits were Zika hybrid rabbits. In the intact does, AMH measurement was complemented by determination of progesterone levels, gynaecological examination and histopathological evaluation of the uterus and ovaries, including follicle counts. Serum AMH and progesterone concentrations were measured using a human-based chemiluminescence immunoassay (CLIA) and an enzyme-linked fluorescence assay (ELFA), respectively. Depending on progesterone levels, sexually intact does were classified into follicular (n=52) or luteal phase (n=12). Median serum AMH levels were 1.53 ng/ml (range 0.77-3.36 ng/ml) in intact and 0.06 ng/ml (range ≤0.01-0.23 ng/ml) in spayed does. AMH concentrations between the intact and spayed rabbits differed significantly and did not overlap (p < .001). Receiver operating characteristic (ROC) curve analysis yielded a sensitivity and specificity of 100% for a cut-off level of 0.50 ng/ml. Follicular or luteal phase had no significant influence on measured AMH levels (t=0.061, df=62, p=.951). While the number of secondary follicles correlated significantly with AMH concentrations (rs =0.410, p=.001), the number of primary or antral follicles did not (rs =0.241, p=.055 and rs =0.137, p=.281, respectively). In conclusion, a single determination of serum AMH concentrations was adequate to distinguish spayed from intact female rabbits. Among sexually intact individuals, whether does were in follicular or luteal phase had no significant influence on measured serum AMH concentrations. The relationship between small growing follicles and AMH levels as described in other species could be partially confirmed, as secondary follicles correlated significantly with AMH.

The use of anti-Müllerian hormone (AMH) as a biomarker for assessment of male gonadal activity has become increasingly widespread. Aberrant AMH concentration successfully detects cryptorchids or pathologic testes in postpubertal stallions, but the ability to use AMH to identify stallions with testicular pathologies during their prepubertal life has not been analysed so far. The objectives of this work were to (1) assess AMH, testosterone, LH, and FSH dynamics in male horses with or without testicular pathologies during the first year of life; and (2) find a reliable diagnostic approach that would enable the identification of animals that will develop abnormal testes at an early stage. Warmblood colts (n = 16) born from February to May on the same stud farm were used. Blood samples for hormone determinations were collected from birth onwards every 4 weeks until the age of 1 year. At 2 years, testicular development was assessed, total testicular volume calculated. and a blood sample collected. Concentrations of AMH, testosterone, LH, and FSH were determined in all samples; AMH (AL-115, Ansh Laboratories, Webster, TX, USA) and testosterone (DE1559, Demeditec, Kiel-Wellsee, Germany) concentrations were determined by ELISA, whereas LH and FSH concentrations were determined by radioimmunoassay. Statistical analysis (SPSS Statistics 24; IBM/SPSS, Armonk, NY, USA) was performed by ANOVA using a general linear model for repeated measures. In 2 stallions, unilateral cryptorchism, and in other 4 stallions, subnormal total testicular volume (&lt;mean minus SD) were diagnosed at 2 years. Concentrations of AMH, testosterone, and FSH changed over time (P &lt; 0.001) but were similar (P &gt; 0.05) within the first year of life irrespective of testicular morphology and location. Concentration of LH at birth was lower (P = 0.05) in stallions with abnormal testes (0.3 ± 0.2 ng mL−1) than in controls (0.6 ± 0.2 ng mL−1), but did not differ thereafter. At 2 years of age, AMH concentration was higher (P &lt; 0.01) in stallions with abnormal testes (39.7 ± 12.7 ng mL−1) than in controls (8.0 ± 0.2 ng mL−1), but no group differences with regard to LH, FSH, and testosterone existed. There was a low but significant negative correlation between AMH and FSH (P &lt; 0.001, r = –0.24), as well as between AMH and LH (P &lt; 0.05, r = –0.15). Also, testosterone concentration was positively correlated with FSH (P &lt; 0.05, r = 0.18) and LH (P &lt; 0.05, r = 0.16) concentrations. In conclusion, AMH determination can be reliably used from 2 years onwards to identify stallions with abnormal testicular development, but it is inconclusive before puberty. We concluded that LH secretion in the perinatal period is involved in testicular development and descent in the horse.

Anti-Müllerian hormone, testosterone, and insulin-like peptide 3 as biomarkers of Sertoli and Leydig cell function during deslorelin-induced testicular downregulation in the dog

Reproduction and culling of productive animals are the main factors that affect the number of cows in a herd. Timely determination of fertility in replacement heifers with subsequent culling of problem heifers will help reduce direct economic losses in breeding dairy cattle. When making a diagnosis, veterinarians should exclude all forms of temporary or permanent infertility, including Congenital anomalies (infantilism, freemartinism, deformities, etc.).In order to establish the hormonal status of heifers-freemartin in the blood serum, the concentration of sex hormones (progesterone, estradiol-17ᵦ, and testosterone) and anti-Müllerian hormone (AMH) was determined.Studies of the level of AMH using an enzyme-linked immunosorbent assay in blood serum have proven the possibility of early assessment of fertility in heifers. This method will allow early culling of Freemartin heifers from a productive herd. The study of AMH levels showed that its amount directly correlates with fertility in cattle, regardless of the period of the estrous cycle. When the concentration of AMH in the blood serum is below 100 pg / ml, heifers are not capable of fruitful insemination, while a high level of anti-Müllerian hormone (more than 380 ng / ml) makes it possible to successfully fertilize animals on the first attempt. All clinically healthy replacements obtained during a single pregnancy were successfully inseminated. At low concentrations of anti-Müllerian hormone, Freemartin heifers were sterile, even after hormonal stimulation (Co-Synx) and sperm doses were administered without clinical manifestation of sexual desire.The introduction of testing for the level of AMH by the ELISA method in the mandatory gynecological clinical examination of cattle in livestock enterprises is expedient and effective.

The wide variation in circulating anti-Müllerian hormone (AMH) concentrations between mares is attributed to differences in antral follicle count (AFC) which may reflect follicular function. There are few data regarding variations in AFC and associated regulatory factors for AMH in the equine follicle during follicular development. To examine molecular and hormonal differences in the equine follicle in relation to variations in AFC and circulating AMH concentrations during follicular development and to identify genes co-expressed with AMH in the equine follicle. Observational study. Plasma AMH concentrations and AFC were determined in 30 cyclic mares. Granulosa cells, theca cells and follicular fluid were recovered from growing (n = 17) or dominant follicles (n = 13). The expression of several genes, known to be involved in folliculogenesis and steroidogenesis, was examined using real-time reverse transcriptase polymerase chain reaction and immunohistochemistry. Intrafollicular oestradiol and AMH concentrations were determined by immunoassay. Within growing follicles, the expression of AMH, AMHR2, ESR2 and INHA in granulosa cells was positively correlated with AFC and plasma AMH concentrations. In addition, the expression of ESR1 and FSHR was positively associated with plasma AMH concentrations. No significant associations were detected in dominant follicles. Furthermore, there was no association between AMH or oestradiol concentrations in follicular fluid and variations in AFC. Finally, the expression of AMH and genes co-expressed with AMH (AMHR2, ESR2 and FSHR) in granulosa cells as well as intrafollicular AMH concentrations decreased during follicular development while intrafollicular oestradiol concentrations increased and were inversely related to intrafollicular AMH concentrations. This study indicates that variations in AFC and circulating AMH concentrations are associated with molecular changes in the growing equine follicle.

Success of assisted reproductive techniques, as determined by the response to hormonal treatments and embryo quality, can successfully be predicted by the concentration of anti-Müllerian hormone (AMH) in plasma of several species. Being able to predict ovarian follicular reserve of prepubertal female horses (fillies) would help to select fertile broodmares and reduce costs associated with animal upkeep. The objectives of this work were to (1) assess AMH dynamics in female horses during the first year of life and (2) determine whether AMH concentrations detected in plasma of prepubertal fillies are correlated with AMH concentrations and antral follicle count (AFC) after puberty. Warmblood fillies (n = 14) born from February to May of 1 year in the same stud were used. Blood samples for AMH determinations were collected from birth onward every 4 weeks up to the age of 1 year. At 2 years, blood samples were collected and AFC was determined by transrectal ultrasonography. The AMH concentrations were determined by ELISA (AL-115, Ansh Laboratories, Webster, TX, USA). Transrectal ultrasonography was used to determine the AFC, which corresponds to the total number of antral follicles detected with ultrasound. Statistical analysis was done with the SPSS Statistics 24 software (SPSS Inc., Chicago, IL, USA). The AMH was detectable in the plasma of all animals from birth onward. At birth, mean AMH concentration was 4.5 ± 1.2 ng mL−1. The AMH concentration increased and peaked between 24 weeks (8.7 ± 4.4 ng mL−1) and 28 weeks (6.7 ± 2.1 ng mL−1) and subsequently decreased again (52 weeks: 3.9 ± 0.9 ng mL−1). Very high variation among individuals during the first year was lost at 2 years of age. The AMH concentration at 2 years was highly correlated with AMH concentration at birth (r = 0.62, P &lt; 0.05) and with AFC (r = 0.78, P &lt; 0.001). Also, AMH concentration (r = 0.73, P &lt; 0.01) and AFC (r = 0.6, P &lt; 0.05) at 2 years were highly correlated with AMH concentrations at 24 and 28 weeks. Gestational length (337 ± 1 days), parity of the dam (4.6 ± 0.8), and placental weight (6983 ± 352 g) did not influence AMH concentrations at any time. Our results demonstrate that AMH is detectable in blood of female horses from birth onward. Despite its high variability between individuals up to 52 weeks, strong correlations were observed during the first 2 years of life. High correlations to AFC at 2 years suggest that determination of AMH in prepubertal female horses helps to predict the ovarian reserve and fertility in postpubertal life.

BackgroundTesticular tumours are common in dogs and in many cases do not give rise to clinical signs. In other cases, signs of feminization, hyperpigmentation or alopecia may be observed, most commonly associated with Sertoli cell tumours (SCT). Although these signs are often associated with elevated concentrations of oestradiol, analysis of oestradiol may give inconclusive results due to large variations among individuals. Other biomarkers are therefore needed. Anti-Müllerian hormone (AMH) is expressed by the Sertoli cell. In humans, AMH has been shown to be a specific marker of Sertoli cell origin in gonadal tumours. Using immunohistochemistry, AMH has been shown to be a useful marker of immature and neoplastic Sertoli cells in dogs. The aim of the present study was to evaluate the clinical relevance of AMH analysis in peripheral blood in the diagnostic workup of dogs with suspected testicular tumours.ResultsBlood was collected from 20 dogs with a palpable testicular mass and from 27 healthy controls. Serum was analysed for oestradiol-17β using a RIA and for AMH using an ELISA. The Mann–Whitney U test was used to compare hormone concentrations between different groups.All control dogs had AMH concentrations ≤ 10 ng/mL, except one outlier that had a concentration of 43 ng/mL. Six dogs with SCT or mixed tumours containing SCT had AMH concentrations higher than 22 ng/mL, significantly higher than AMH concentrations in control dogs (P = 0.0004). Concentrations between 10 and 22 ng/mL were found in about half of the dogs with non-neoplastic testicular pathologies or with testicular tumours other than SCTs. Age did not significantly affect concentrations of AMH in the control dogs.ConclusionAMH was shown to be a promising biomarker for the diagnosis of Sertoli cell tumours in dogs.

Anti-Mullerian hormone (AMH), also called Mullerian-inhibiting substance or factor, is a dimeric glycoprotein, produced by immature Sertoli cells, and responsible for Mullerian regress ion in male fetuses. To study the ontogeny of AMH production in the male against human recombinant AMH, the AMH concentration in 21 samples of amniotic fluid and 44 samples of fetal serum, initially collected for cytogenetical analysis in fetuses with sonographic abnormalities. No AMH was detectable in amnitotic fluid, whatever the fetal sex. Mean ± SEM AMH concentration was 40.5 ± 3.9 ng/ml in the serum of male fetuses from 19 to 30 weeks, and 28.4 ± 6.1 ng/ml in older ones. The AMH concentration in the serum of an male XX fetus, aged 24 weeks, 48.3 ng/ml, was the only biological indicator of fetal sex. No AMH was detectable in female serum at any time, allowing easy discrimination between male and female samples, even during the perinatal period, when mean scrum AMH concentration is decreased, compared to that of infants aged 2 months to 2 years (43.1 ± 3.7? P>0.05). AMH production in early fetal life was studied by in-situ hybridization, using AMH-specific sense and antisense riboprobes. AMH transcripts were detected in the Sertoli cells of fetuses aged 8 weeks or older, but not in ovarian tissue. Negative results were also found in the sexually undifferentiated gonadal tissue of one 7-week-old fetus, with detectable DMA SRY-specific sequences, confirming that AMH expression in the testis begins only after seminiferous tubule differentiation.

What is the relationship between the anti-Müllerian hormone (AMH), gonadotropin and androgen concentrations within a single follicle and live birth after ICSI and a transfer of an embryo developed from the matched oocyte? Among the analysed markers on the day of oocyte retrieval, AMH concentration in follicular fluid (FF) is a predictor of live birth after single embryo transfer (SET). High serum concentrations of AMH and low FSH concentrations have been associated with a high chance of pregnancy after ART. Whether there are differences in the hormonal milieu for individual follicles and whether this impacts the laboratory and clinical outcomes for the individual oocyte developing within that follicle are unknown. This prospective cohort study included 322 individual FF samples from 199 infertile women scheduled for ICSI/SET over an 18-month period. Of these women, 76 provided a single FF sample, while 123 women contributed two FF samples taken from two different follicles. The first follicle aspirated in each ovary on the day of oocyte retrieval had the FF aspirated; the individual cumulus-oocyte complex (COC) was tracked, and the associated FF was stored at -80°C. FF AMH, FSH, LH, testosterone (T) and androstenedione (A2) levels were measured by mass spectrometry (androgens) and immunoassays. The laboratory and clinical outcomes for each individual oocyte were related to their unique follicle hormone concentrations. Of the 322 oocytes with paired FF samples, 70 (21.7%) oocytes did not fertilise. From the remaining 252 2PN embryos, 88 (34.9%) were transferred as single embryos on Day 3; of the remaining 164, 78 developed into blastocysts, and 18 single blastocyst transfers were performed. Thus, a total of 106 transferred embryos had matching FF samples. An analysis of these individual FF concentrations revealed that AMH concentrations were higher in follicles in which the oocyte developed into a top quality (TQ) blastocyst (6.33 ± 5.52ng/ml) and whose transfer led to live birth (7.49 ± 5.03ng/ml) than those in which there was a failure of fertilisation (3.34 ± 2.21ng/ml). In contrast, follicular FSH concentrations were the lower for oocytes that resulted in a TQ blastocyst (5.36 ± 2.20 mIU/ml) and live birth (5.60 ± 1.41 mIU/ml) than for oocytes that failed to fertilise (9.06 ± 3.36 mIU/ml). FF AMH was the only studied marker that increased the chance of live birth (odds ratio: 1.93 [95% CI: 1.40-2.67], P < 0.001). The receiver operating characteristic analysis showed that FF AMH levels predicted live birth with a very high sensitivity (91.2%), specificity (91.7%) and an excellent AUC value of 0.954, whereas serum AMH level only had a fair (AUC = 0.711) significance as a predictor for live birth after ICSI/SET. The predictive capabilities of the interfollicular markers were not limited to the TQ embryos or blastocysts; they applied to all SET cycles. Whether an altered intrafollicular hormonal environment reflects the developmental capacity of the oocyte or defines cannot be determined from this cross-sectional analysis. Inclusion of 21 subjects with polycystic ovary syndrome (PCOS) may have biased the findings due to a unique intrafollicular milieu associated with PCOS. Our results suggest that highly competent human oocytes have an FF composition of AMH, FSH, T and A2 that is close to that in a natural cycle. Also, the relationships between intrafollicular AMH, gonadotropin and androgen levels in the same follicle support the hypothesis that FF AMH concentration may reflect granulosa cell proliferation during gonadotropin-stimulated follicle growth. Finally, the serum AMH concentration is markedly lower than the FF AMH concentration, with a moderate correlation between serum and FF AMH, implying ovarian follicle autonomy with regards to its secretory products. The National Science Centre of Poland supported this work (grant number: N N407 217 040). The authors declare that there is no conflict of interest regarding the publication of this article.

Anti-Mullerian hormone (AMH) concentration in follicular fluid and mRNA expression of AMH receptor type II and LH receptor in granulosa cells as predictive markers of good buffalo (Bubalus bubalis) donors

Changes in serum testosterone and anti-Müllerian hormone concentration in bulls undergoing scrotal insulation