Plasma anti-Müllerian Hormone Research Articles

Anti Mullerian Hormone: A Putative Endocrine Marker for Prediction of Superovulation Response in Cattle

Anti-Müllerian Hormone (AMH) is a dimeric glycoprotein molecule with a molecular weight of 140-kDa linked to the Transforming Growth Factor-β (TGF-β) superfamily. Research on the use of AMH in livestock has gained momentum in recent years. In particular, it is now widely used in cattle breeding, where embryo transfer technology is used to obtain more offspring from genetically superior females. One of the most important factors that increase the success of embryo transfer is the response of the selected donor to the superovulation protocol. AMH has been successfully used as a biomarker in predicting superovulation response in cattle and in estimating the numbers of oocytes collected by ovum pick up (OPU). AMH plasma concentrations are positively and highly correlated with antral follicle count (AFC) in cattle and can also be used as a marker of ovarian reserve. In addition, AMH was also positively and highly correlated with the number of corpus luteum (CL) and total embryos after superovulation in several studies. It has been also reported via Genome-Wide Association Studies (GWAS) that plasma AMH level is an inherited trait in cattle and can be improved through genomic selection. In this study, we aimed to evaluate the relationship between plasma AMH levels and superovulation response in cattle by compiling the data obtained from various studies in light of current scientific literature.

Assessment of semen parameters and Anti-müllerian hormone level in patients with chronic kidney disease on hemodialysis: a case control study

Background and Objective: Chronic kidney disease (CKD) is a common health problem which is accompanied by considerable morbidity and mortality. Anti-müllerian hormone (AMH) is one of the hormones that are affected by CKD. The objective was to assess semen parameters and AMH level in CKD patients on hemodialysis. Material and methods: This study was a case control study of 20 patients with stage 4 and 5 CKD on hemodialysis, attending Nephrology Outpatient Clinic and Renal Dialysis Unit of Suez Canal University hospital and 20 healthy fertile controls. Full history, clinical examination, kidney function tests and semen analysis were performed. AMH level in serum and seminal plasma was measured using ELISA. Results: The mean AMH level in serum of case group was 3.35 ± 2.43 ng/mL versus 4.13 ± 3.17 ng/mL in the controls respectively (p > 0.05) while mean AMH level in semen of the case group was 2.13 ± 2.64 ng/mL compared to 9.37 ± 3.79 ng/mL in controls respectively (p < 0.05). There was a positive correlation between AMH in seminal plasma and sperm count, total sperm motility, progressive sperm motility and sperm viability, while the AMH level in serum was insignificantly related. Conclusion: Semen parameters in CKD patients on hemodialysis were significantly poorer than normal controls AMH in seminal plasma was significantly decreased in CKD patients (stage 4–5 on hemodialysis) and significantly correlated with the semen parameters.

Skin Mediates the Aphrodisiac Effect of UVB Light Via a Skin-Brain-Gonad Axis

Ultraviolet (UV) light affects endocrinological and behavioral aspects of human sexuality via an unknown mechanism. Using a unique male-female comparative approach, we discovered that the sexual behavioral and hormonal features enhanced by UVB are mediated by the skin. In mice, UV exposure increases hypothalamus-pituitary-gonadal axis hormone levels, resulting in enhanced ovary size, extended estrus days, and anti-Mullerian hormone (AMH) expression. It likewise enhances sexual responsiveness and attractiveness of females and heterosexual interactions of both males and females. Conditional knockout of p53 specifically in skin keratinocytes abolished UV’s effects. In humans, UV exposure enhanced romantic passion in both genders, increased testosterone levels in men and correlated with heightened plasma AMH levels in women. Our data, revealing that UVB triggers a skin-brain-gonadal axis through skin p53 activation, offers therapeutic opportunities for sex-steroid-related dysfunctions. We speculate that during human furless skin evolution, the skin became the front-line regulator of the response to UVB.

OVARIAN AGING AND REPRODUCTIVE SENESCENCE IN MOUSE MODEL OF MITOCHONDRIAL DYSFUNCTION

To evaluate the mechanisms of female reproductive aging and senescence induced by mitochondrial dysfunction in mice. Prospective cohort design study. 13 female mice with mitochondrial dysfunction (mtDNA-depleter) and 13 wild-type (WT) female mice were studied at three time points in their reproductive life cycle: 4-6, 8-10, and 11-12 months of age. A transgenic mouse model with an inducible gene for mitochondrial dysfunction (mtDNA-depleter) was previously created by our lab. All protocols were approved by our institutional IACUC. At each of the three time points, reproductive markers were assessed, including estrous cyclicity, plasma anti-Mullerian hormone (AMH), ovarian follicles, estrogen receptor staining, and uterine and vaginal atrophy. Breeding experiments were performed to assess fertility. Differences between the mtDNA-depleter and WT groups were evaluated by independent two-sample t-test. Twenty-six female mice were studied from 4 to 12 months of age. The estrous cycles in 4-6-month mtDNA-depleter mice were prolonged, 9.4 days (n = 7), versus 5.3 days in WT (n = 9), p-value of 0.021. By 11-12 months, both groups had prolonged estrous cycles consistent with physiologic aging. AMH levels in 4-6-month mtDNA-depleter mice were decreased, 84 (n = 4), versus 114 ng/mL in WT (n = 5), p-value of 0.013. AMH was reduced in older mice at 8-10 and 11-12 months, but there were no significant differences between groups. Looking at the data collectively for all three time points, there were fewer tertiary (early antral, antral, or pre-ovulatory) follicles in mtDNA-depleter mice than in WT, 3.5 versus 4.9 per cut section, respectively (p-value of 0.039). There were fewer corpora lutea in mtDNA-depleter mice than in WT, 0.90 versus 1.7 per cut section, respectively (p-value of 0.027). Collectively, average uterine weight in mtDNA-depleter mice was lower, 62.2, versus 92.3 mg in WT mice, p-value of 0.0028. Vaginal epithelium in 4-6-month mtDNA-depleter mice was thinner, 98 μm (n = 4), versus 200 μm in WT mice (n = 4), p-value of 0.035. In breeding experiments, mtDNA-depleter mice at age 3 and 5 months did not produce litters. Estrogen receptor levels were reduced in mtDNA-depleter mouse ovary relative to WT, evident from both immunofluorescence and semi-quantitative PCR analysis. Mitochondrial dysfunction in mice is associated with premature reproductive aging and senescence, as evidenced by early onset of prolonged estrous cycles, diminished ovarian reserve, uterine and vaginal atrophy, and sterility. Mitochondrial dysfunction is also associated with premature estrogen receptor down-regulation in the mouse ovary.

Anti-Mullerian hormone and its relationship to ovulation response and fertility in timed AI Bos indicus heifers.

This study evaluated the association between plasma anti-Mullerian hormone (AMH) concentration and fertility in Nelore (Bos indicus) heifers submitted to timed artificial insemination (TAI). At the onset of the synchronization protocol, heifers (n=289) received a subcutaneous P4 ear implant (3mg) and 2mg of oestradiol benzoate. Eight days later, the P4 implant was removed and 0.5mg of oestradiol cypionate, prostaglandin (0.265mg, i.m.) and equine chorionic gonadotropin (300UI, i.m.) was administered, and TAI was performed 48hr after ear implant removal. Ovarian ultrasound evaluations were performed to measure number of ovarian follicles, dominant follicle size and ovulation response. Pregnancy diagnosis was performed by ultrasound 30days after AI. Heifers with greater circulating AMH had more antral follicles, a smaller dominant follicle near timed ovulation and lower ovulation response to the timed AI protocol compared to heifers with lower circulating AMH. Although AMH and pregnancy outcome had a quadratic-shaped pattern, AMH was not significantly associated with fertility. In conclusion, heifers with lower AMH had larger follicles towards the end of the synchronization protocol and greater ovulation responses, whereas greater circulating AMH was unrelated to conception success.

CORRELATION BETWEEN PLASMA LEAD AND SERUM ANTI MULLERIAN HORMONE LEVELS IN WOMEN WITH PREMATURE OVARIAN FAILURE

Background: Lead is a highly toxic metal to humans, with the injurious effects on the hemopoietic, nervous and reproductive system. Lead crosses the placenta during pregnancy and has been associated with intrauterine death, prematurity and low birth weight. Objective: To collate between plasma lead and serum antimullerian hormone levels in women with premature ovarian failure, by estimation of level of Anti Mullerian Hormone (AMH), Follicular Stimulating Hormone (FSH), Estradiol (E2) and plasma Lead level. Subjects and Method: This study was conducted in Al Azhar University Hospital outpatient clinics after approval of the research and ethical committee during the period from December 2015 to December 2018. Study groups were: Group 1 included 50 normal candidate women under 40 years old has regular menstrual cycles (21-35 days), fertile (at least one natural pregnancy carried to term), and serum FSH was less than 10 mlU/ml, Group 2 included 50 patients with premature ovarian failure who aged under 40 years with absence of spontaneous menstrual bleeding for more than 12 months, serum estradiol level was less than 25pg/ml, serum FSH level more than 40 mlU/ml. AMH, plasma lead level in patients with premature ovarian failure diagnosed and correlated with level of serum FSH with the normal control group. Results: There was a statistically significant positive correlation between plasma lead levels with FSH among the study group. On the other hand, there was no statistically significant correlation between plasma lead level and AMH level. Conclusion: The anti mullerian hormone showed a significant difference between the cases of premature ovarian insufficiency and the control group. Moreover, AMH was a good marker for ovarian reserve and ovarian response for gonadotropins simulation. Chronic exposures to heavy metals, especially lead, affected the ovarian function and impairment in folliculogenesis which increased the incidence of Premature Ovarian Failure (POF) especially in rural area due to the low socioeconomic status.

CORRELATION BETWEEN PLASMA LEAD AND SERUM ANTI MULLERIAN HORMONE LEVELS IN WOMEN WITH PREMATURE OVARIAN FAILURE

Background: Lead is a highly toxic metal to humans, with the injurious effects on the hemopoietic, nervous and reproductive system. Lead crosses the placenta during pregnancy and has been associated with intrauterine death, prematurity and low birth weight. Objective: To collate between plasma lead and serum antimullerian hormone levels in women with premature ovarian failure, by estimation of level of Anti Mullerian Hormone (AMH), Follicular Stimulating Hormone (FSH), Estradiol (E2) and plasma Lead level. Subjects and Method: This study was conducted in Al Azhar University Hospital outpatient clinics after approval of the research and ethical committee during the period from December 2015 to December 2018. Study groups were: Group 1 included 50 normal candidate women under 40 years old has regular menstrual cycles (21-35 days), fertile (at least one natural pregnancy carried to term), and serum FSH was less than 10 mlU/ml, Group 2 included 50 patients with premature ovarian failure who aged under 40 years with absence of spontaneous menstrual bleeding for more than 12 months, serum estradiol level was less than 25pg/ml, serum FSH level more than 40 mlU/ml. AMH, plasma lead level in patients with premature ovarian failure diagnosed and correlated with level of serum FSH with the normal control group. Results: There was a statistically significant positive correlation between plasma lead levels with FSH among the study group. On the other hand, there was no statistically significant correlation between plasma lead level and AMH level. Conclusion: The anti mullerian hormone showed a significant difference between the cases of premature ovarian insufficiency and the control group. Moreover, AMH was a good marker for ovarian reserve and ovarian response for gonadotropins simulation. Chronic exposures to heavy metals, especially lead, affected the ovarian function and impairment in folliculogenesis which increased the incidence of Premature Ovarian Failure (POF) especially in rural area due to the low socioeconomic status.

Abstract P132: Anti-Müllerian Hormone Levels and Risk of Type 2 Diabetes in Women

Introduction: Given its role in ovarian follicle development, circulating anti-Müllerian hormone (AMH) is correlated with timing of menopause. Accordingly, women with higher AMH levels become menopausal at a higher age. Previous research suggests that a higher age at menopause is associated with a decreased risk of type 2 diabetes (T2D). In contrast, AMH levels are increased in women with polycystic ovary syndrome (PCOS), who have a higher risk of insulin resistance and T2D than women without PCOS. However, it is not clear yet whether AMH actually plays a role in the development of T2D. We aimed to investigate whether plasma AMH levels and age-related AMH trajectories are associated with risk of T2D in women. Hypothesis: Higher age-specific plasma AMH levels are associated with a decreased risk of T2D. Methods: We analyzed longitudinal data from 3104 female participants, aged 20-60 years at recruitment, in the population-based Doetinchem Cohort Study. In total, we analyzed 12460 plasma AMH measurements. We calculated age-specific AMH tertiles, to account for the strong AMH-age correlation. Cox Proportional hazards models adjusted for known risk factors for diabetes were used to assess the relation between age-specific AMH tertiles and T2D. We applied linear mixed models to compare age-related AMH trajectories between T2D cases and non-cases. Results: After a median follow-up of 20 years, 163 incident T2D cases were identified. Higher age-specific AMH levels were associated with a lower risk of T2D (hazard ratio (HR) T2vsT1 = 0.77, 95%CI: 0.53-1.11; HR T3vsT1 =0.62, 95%CI: 0.40-0.94; p for trend = 0.02). These findings were supported by predicted AMH trajectories, which suggested that plasma AMH levels were lower at younger ages and declined at a slower rate in women who were diagnosed with T2D compared to women who were not. However, differences in trajectories between T2D cases and non-cases were not statistically significant. Conclusions: We observed that women with higher age-specific AMH levels were at a lower risk of T2D. In addition, our longitudinal analyses may suggest that AMH is lower in women who develop T2D compared to women who do not. These findings are in line with previous studies that observed that a higher age at menopause was associated with a decreased risk of T2D.

Efficacy of a single blood anti-Müllerian hormone (AMH) concentration measurement for the selection of Japanese Black heifer embryo donors in herd breeding programs.

We evaluated the relationship between plasma anti-Müllerian hormone (AMH) concentrations in Japanese Black (JB) heifers at 7−10 months of age and the number of embryos recovered after superovulation treatment in selected ovum pick-up donors, concomitantly with changes in their AMH concentrations before and after parturition. Plasma AMH concentrations in heifers were positively correlated with the total number of follicles (r = 0.647, P < 0.01) and embryos (r = 0.681, P < 0.01) recovered from the animals postpartum, when selected as donor cows, but did not correlate with the total number of transferable embryos. No difference was observed between the plasma AMH concentration at the heifer period and the postpartum period. Additionally, serum AMH concentrations of heifers weakly correlated with the number of follicles and embryos recovered by virgin flush after superovulation treatment at 13−15 months of age. Therefore, a single blood AMH concentration measurement may accelerate intensive JB cattle breeding.

Neonatal Exposure to Agonists and Antagonists of Sex Steroid Receptors Affects AMH and FSH Plasma Level and Their Receptors Expression in the Adult Pig Ovary.

Simple SummaryThe ovarian development and the establishment of ovarian reserve during fetal and/or neonatal life is critical for future reproductive success. Many environmental chemicals are known to negatively affect development and physiology of human and animal ovaries by interfering with endocrine systems, resulting in aberrant reproductive functions. The present study shows the long-term impact of neonatal exposure to agonists and antagonists of sex steroid receptors on AMH and FSH signalling in the ovary of adult pigs. Our findings suggest alteration in ovarian follicle recruitment from ovarian reserve arising from neonatal disruption of androgen/estrogen signalling induced by environmental endocrine active compounds. Everyday use of many endocrine disruptors is already prohibited after their harmful impacts on normal physiology have become known. Nevertheless, market introduction of new chemicals with potential deleterious influence on reproductive physiology has continued. Our outcomes confirm that a neonatal window plays an essential role in the physiological programming of ovarian function in adult pigs. The influence of environmental chemicals on this critical neonatal window needs to be investigated in order to gain a comprehensive view of deleterious interactions between endocrine disrupting chemicals and ovarian function.In this study piglets were injected with testosterone propionate (TP, an androgen), flutamide (FLU, an antiandrogen), 4-tert-octylphenol (OP, an estrogenic compound), ICI 182,780 (ICI, an antiestrogen) or corn oil (controls) between postnatal days 1 and 10 (N = 5/group). Then plasma anti-Müllerian hormone (AMH) and follicle stimulating hormone (FSH) concentration and the expression of their receptors were examined in the adult pig ovary. TP and FLU decreased plasma AMH and FSH concentration. In preantral follicles, TP resulted in upregulation of AMHR2 and FSHR expression, but decreased AMH protein abundance. FLU upregulated AMHR2 expression, while OP increased FSHR mRNA. In small antral follicles, OP upregulated ACVR1 and BMPR1A expression, while FLU increased BMPR1A mRNA. FLU and ICI resulted in upregulation of AMHR2 expression. TP and FLU upregulated AMH expression, while it was downregulated in response to OP or ICI. Moreover, OP and ICI resulted in downregulation of FSHR expression, while FLU decreased FSHR protein abundance. In conclusion, neonatal exposure to either agonist or antagonist of androgen receptor affected AMH and FSH signalling systems in preantral follicles. In small antral follicles these systems were influenced by compounds with estrogenic, antiestrogenic, and antiandrogenic activity. Consequently, these hormonal agents may cause an accelerated recruitment of primordial follicles and affect the cycling recruitment of small antral follicles in pigs.

AMH: Could It Be Used as A Biomarker for Fertility and Superovulation in Domestic Animals?

Anti-Müllerian hormone (AMH) is a reliable and easily detectable reproductive marker for the fertility competence of many farm animal species. AMH is also a good predictor of superovulation in cattle, sheep, and mares. In this review, we have summarized the recent findings related to AMH and its predictive reliability related to fertility and superovulation in domestic animals, especially in cattle. We focused on: (1) the dynamics of AMH level from infancy to prepubescence as well as during puberty and adulthood; (2) AMH as a predictor of fertility; (3) the association between antral follicle count (AFC) and plasma AMH level; (4) AMH as a predictor of superovulation; and (5) factors affecting AMH levels in domestic animals, especially cattle. Many factors affect the circulatory levels of AMH when considering the plasma, like nutrition, activity of granulosa cells, disease state and endocrine disruptions during fetal life. Briefly, we concluded that AMH concentrations are static within individuals, and collection of a single dose of blood has become more popular in the field of assisted reproductive technologies (ART). It may act as a potential predictor of fertility, superovulation, and ovarian disorders in domestic animals. However, due to the limited research in domestic animals, this potential of AMH remains underutilized.

224 Superstimulation effect on invitro embryo production and relationship with anti-Müllerian hormone in buffaloes (Bubalus bubalis) aspirated 7 or 14 days after follicular ablation

The success of buffalo invitro embryo production (IVEP) depends, in part, on the number and competence of oocytes. Superstimulation with FSH has been linked to oocyte quality and embryo yield, and anti-Müllerian hormone (AMH) has been proposed as an indicator of ovarian reserve. The objective was to evaluate FSH stimulation in one ovum pick-up (OPU) cycle conducted 7 or 14 days after follicular ablation; we additionally analysed residual FSH treatment effect carried over to a subsequent aspiration cycle. Finally, we evaluated the relationship between plasma AMH and FSH response. Nine multiparous, cycling Murrah donors (603±7.0kg, body condition score=3.5) were subjected to complete follicular ablation followed by OPU 1 week later (T0 Control), and plasma AMH was determined at this point. They were then randomly allocated to FSH stimulation (3 days b.i.d. in descending doses, total 300mg of pFSH, Vetpharm, with the last injection 48h before OPU) with aspiration conducted 7 (T7 FSH, n=5) or 14 days (T14 FSH, n=4) post-follicular ablation. The residual effect (T Residual, n=5) was evaluated in one subsequent, nonstimulated OPU cycle conducted 1 week later (9 days after last FSH administration). For plasma AMH, concentration was measured using ELISA kit (cat. no. AL114, AnshLabs; sensitivity 1.0 pgmL−1, interassay variation &amp;lt;5%, additional validation conducted with serial dilutions of buffalo plasma, r=0.98 against standard curve). Continuous data were analysed with ANOVA and Tukey's post hoc comparisons; categorical data were analysed using Chi-squared test. Pearson coefficient was used to evaluate correlation between AMH, follicular reserve, and invitro performance. Superovulatory treatment resulted in a higher number of follicles and good quality oocytes (grades 1 and 2) per donor, irrespective of OPU interval (T7 FSH: 3.0±2.55 and T14 FSH: 1.5±0.5 vs. T0 Control: 0.45±0.38 and T Residual: 0.4±0.4; P&amp;lt;0.05). There were more cleaved embryos (T7 FSH: 26%, T14 FSH; 30% vs. T0 Control: 7% and T Residual: 20%) and blastocysts (T7 FSH: 21%, T14 FSH; 30% vs. T0 Control: 5% and T Residual: 0; P&amp;lt;0.05) from FSH-treated donors. There was a strong correlation (r=0.9439) between follicle count and plasma AMH, and weaker relationships for oocytes recovered (r=0.3975), cleaved embryos (r=0.2317), or blastocysts (r=0.0106). Results indicate that FSH treatment with a 48-h coasting period results in significant improvements in follicular population, oocyte quality, and embryonic development; these effects are observed independently of the interval between initial follicular ablation and OPU. No residual effects of FSH treatment were observed, indicating that the beneficial effects of superstimulation are not carried over to the following OPU cycle. In conclusion, a strong correlation coefficient was established between initial plasma AMH level and follicular response to FSH treatment, indicating that it may be used as a selection criterion for animals likely to exhibit a good response to superstimulatory treatment.

157 Effect of a slow-release gonadotrophin-releasing hormone analogue on ovarian activity and oestrous behaviour in mares

Oestrus behaviour in mares can contribute to problems in their handleability and reduced performance in equestrian sports. Therefore, methods of transient suppression of oestrous cyclicity in mares are of interest. The aim of our study was to determine whether treatment of mares with slow-release implants containing the gonadotrophin-releasing hormone (GnRH) analogue deslorelin downregulates pituitary GnRH receptors and reduces ovarian function and oestrous behaviour. Shetland mares (age=11.0±1.4 years; bodyweight=185.5±7kg) were oestrous synchronised with two injections of the prostaglandin F2α analogue luprostiol (3.725mg) at an interval of 12 days. One day after the second injection (Day 0), mares were randomly assigned to three groups: slow-release implant with 9.4mg of deslorelin (Suprelorin, Virbac; group D1; n=6), implant with 4.7mg of deslorelin (group D2; n=5), and intramuscular injection of 1.25mg of short-acting deslorelin (control, group C; n=5). Collection of blood samples for analysis of progesterone, LH, and anti-Müllerian hormone (AMH) using established and validated enzyme immunoassays (Scarlet et al. 2018 Theriogenology 117, 72-77), testing for oestrus-like behaviour with a Shetland stallion, and ultrasonography of the genital tract were performed at 2-day intervals until Day 10 after treatment and at 5-day intervals from there. On Days 10 and 45 after treatment, LH stimulation tests with the GnRH agonist buserelin (4µg IV) were performed. Data were normally distributed; differences among groups were analysed using analysis of variance and subsequent Tukey test. Values are means±s.e.m. In all mares without a corpus luteum on Day 0 (progesterone &amp;lt;1ngmL−1; one mare in group D1 and two in group C), ovulation was detected within 9 days after deslorelin treatment. These ovulations were classified as deslorelin induced, whereas ovulations after Day 10 were classified as spontaneous ovulations. The mean interval from deslorelin until the first spontaneous ovulation was 62.0±8.6, 44.2±14.1, and 22.2±3.1 days in groups D1, D2, and C (P&amp;lt;0.05), respectively. Subsequent oestrous cycles were regular. Oestrus-like behaviour until day 50 was reduced in groups D1 (2.0±0.9 days) and D2 (2.4±1.3 days) compared with group C (6.4±1.2 days; P&amp;lt;0.05). Concentration of plasma LH and AMH decreased in group D1 (P&amp;lt;0.05) but not in groups D2 and C. The GnRH stimulation test on Day 10 resulted in an increase (P&amp;lt;0.001) in plasma LH concentration in group C but not in groups D1 and D2 (treatment×time P&amp;lt;0.05). On Day 45, LH concentration increased in all mares in response to buserelin (NS among groups). Within 100 days of treatment, LH concentrations but not AMH concentrations in mares of group D1 returned to baseline. In conclusion, deslorelin slow-release implants transiently suppress ovarian function and oestrus behaviour in mares. Spontaneous ovulation is delayed in a dose-dependent manner. A decrease in AMH concentration suggests inhibitory effects of deslorelin on small antral follicles. Long-term effects on follicular dynamics and fertility in larger horses also need to be assessed.

Anti-Müllerian hormone in grazing dairy cows: Identification of factors affecting plasma concentration, relationship with phenotypic fertility, and genome-wide associations

The objectives of this study were to (1) characterize the distribution and variability of plasma anti-Müllerian hormone (AMH) concentration; (2) evaluate factors associated with phenotypic variation in plasma AMH; (3) examine the associations between categories of plasma AMH and reproductive outcomes [pregnancy to first artificial insemination (P/AI), and pregnancy rates within 21, 42, and 84 d after the mating start date (MSD)]; (4) estimate pedigree and genomic heritability for plasma AMH; and (5) identify and validate SNP associated with phenotypic variation in plasma AMH. Plasma AMH concentration (pg/mL) was determined from a blood sample collected (mean ± standard deviation) 10 ± 2 d after first insemination at detected estrus (IDE) in 2,628 first- and second-parity Irish dairy cows. Overall, plasma AMH had a positively skewed distribution with mean (± standard deviation), median, minimum, and maximum concentrations of 326 ± 231, 268, 15, and 2,863 pg/mL, respectively. Plasma AMH was greatest for Jersey, followed by Holstein × Jersey, Holstein × Norwegian Red, and Holstein cows (410, 332, 284, and 257 pg/mL, respectively). Second-parity cows had greater plasma AMH than first-parity cows (333 vs. 301 pg/mL, respectively). Samples collected at 7 and 8 d after first IDE had lesser plasma AMH than those collected on d 9, 10, 11, 12, and 13 after first IDE (291 and 297 vs. 317, 319, 331, 337, and 320 pg/mL). Plasma AMH was not associated with either body condition score at first IDE or the interval from calving to MSD. Cows were categorized into low (≤150 pg/mL; n = 526; lowest 20%), intermediate (>150 to ≤461 pg/mL; n = 1,576; intermediate 60%), and high AMH (>461 pg/mL; n = 526; highest 20%) groups based on plasma AMH, and associations with reproductive outcomes were tested. Cows with high and intermediate plasma AMH had 1.42- and 1.51-times-greater odds of becoming pregnant within 84 d after the MSD than those with low plasma AMH (90.3 and 90.8 vs. 86.8%, respectively); however, P/AI and pregnancy rate within 21 and 42 d after the MSD did not differ among AMH categories. Plasma AMH was moderately heritable (pedigree heritability of 0.40 ± 0.06 and genomic heritability of 0.45 ± 0.05), and 68 SNP across Bos taurus autosomes 7 and 11 were associated with phenotypic variation in plasma AMH. Out of 68 SNP, 42 were located in a single quantitative trait locus on Bos taurus autosome 11 that harbored 6 previously identified candidate genes (NR5A1, HSPA5, CRB2, DENND1A, NDUFA8, and PTGS) linked to fertility-related phenotypes in dairy cows.

Association of Anti-Müllerian Hormone concentrations between the pregnancy rates and pregnancy continuity of cows in different age groups

The potential relationships between Anti-Müllerian Hormone (AMH) concentrations and fertility were investigated by examining pregnancy rates and early pregnancy loss in different age groups of cows. Holstein heifers (17.35 ± 1.35 months, n = 20), young cows (3.25 ± 1.02 years, n = 20), and old cows (6.7 ± 0.80 years, n = 20) were synchronised and time fixed inseminated. A single blood sample per animal was taken during oestrus just before artificial insemination (AI), (Day 0) for the AMH analysis. The highest plasma AMH concentrations were determined in the heifer group (149.01 ± 12.62 pg/ml, P &amp;lt; 0.001) in the study. The AMH concentrations and conception rates decreased with age. The Day-0-AMH concentrations were higher in the pregnant animals at Day 20 after AI than in the non-pregnant animals (P &amp;lt; 0.05) in each group. The mean Day-0-AMH concentrations in the pregnant animals at Day 60 after AI was higher (P &amp;lt; 0.001) than the AMH concentrations in those with a detected pregnancy loss between D&lt;sub&gt;20&lt;/sub&gt; and D&lt;sub&gt;60&lt;/sub&gt;, and also in those with a pregnancy loss and not being pregnant by Day 20 considered together. When each group was assessed on its own, based on the pregnancy results of Day 60, the Day-0-AMH concentrations were found to be significantly higher in the pregnant heifers than in the non-pregnant heifers and heifers with a pregnancy loss (P &amp;lt; 0.05). In conclusion, the AMH analysis might provide valuable information on the reproductive efficiency of the animals in a herd.

SAT-194 Correlation of Capillary and Venous Plasma Measurements of Anti-Müllerian Hormone

Anti-Müllerian Hormone (AMH) is a member of the transforming growth factor β superfamily which regulates gonadal development and differentiation. AMH is a biomarker for ovarian age and reserve used to assess fertility and predict ovulation induction outcome. It is also a marker for diagnosis and treatment of polycystic ovarian syndrome (PCOS), menopausal transition, and gonadal function. As diagnostic testing begins to expand into self-collection and point-of-care capabilities, there is a need to minimize sample volume requirements and include additional sample types. The TAP device, manufactured by Seventh Sense Biosystems (Medford, MA), is a small, easy to use, single push device that collects approximately 100 µL whole blood from the upper arm. As the blood travels through the channels of the device it is mixed with lithium heparin anticoagulant. The device collects capillary blood and is FDA approved for use in hemoglobin A1c measurements. In this study, AMH levels in venous and capillary plasma were compared. AMH was measured by an electrochemiluminescent immunoassay requiring only 40 µL plasma. The performance characteristics of the assay including accuracy, precision, linearity, and limits of quantitation were assessed. The stability of samples stored in the TAP device was also evaluated. Twenty-one female volunteers ages 24 to 67 donated paired venous and capillary blood samples. Venous plasma from donors with undetectable AMH levels was used as a matrix for spike and recovery studies to verify acceptable accuracy and precision across the analytical measurement range (AMR) and establish the lower (LLOQ) and upper (ULOQ) limits of quantitation. The ULOQ and LLOQ were 15 and 0.15 ng/mL, respectively. Accuracy was within 20% and precision within 15% across the AMR. Linearity was evaluated by serially diluting samples with high levels of AMH in AMH-negative pooled plasma. The maximum acceptable dilution was 8-fold. Correlation of venous and capillary results was excellent. A linear least square model resulted in a coefficient of determination (R2) of 0.998. Four capillary blood samples were collected from each of three donors for stability analysis. One was processed immediately and used as the baseline. The remaining three samples were stored at 2-8°C in the TAP device for 3, 5 and 7 days before processing. Plasma collected after 5 and 7-day storage in the TAP device was moderately hemolyzed and smaller volumes were retrieved. The hemolysis did not impact AMH measurement. AMH was stable at all time points tested with mean recovery of 104% compared to baseline values. This study shows that very small volumes of venous and capillary plasma can be used to accurately measure AMH levels. The ability to measure AMH in capillary blood via the TAP device allows for quick and reproducible blood collection, potentially outside the healthcare setting, improving access to diagnostic information.

A bovine-specific FSH enzyme immunoassay and its application to study the role of FSH in ovarian follicle development during the postnatal period

The primary aim of this study was to develop a FSH enzyme immunoassay (EIA) for the bovine species. The newly developed EIA was validated for FSH determination in bovine plasma by comparison with an existing bovine FSH radioimmunoassay. The EIA detected bovine FSH with a high sensitivity (0.1 ng/ml). Cross-reactivity of the EIA was 0.01% with bovine LH, 51% with ovine FSH, <0.1% with porcine FSH and <0.01% with equine FSH. Using this EIA on different time series of plasma in cows, we have confirmed the presence of a FSH pre-ovulatory peak at estrus, of periodic FSH fluctuations accompanying the waves of terminal follicular development, and of FSH pulses, mainly asynchronous with LH ones, in the peri-ovulatory phase of the cycle. In a second objective, the EIA was used to assess the role of FSH in regulating the development of ovarian follicles up to the small antral stage in young calves. To answer this question, six calves were submitted to weekly blood sampling during their first 3 months of life, and FSH changes were studied concomitantly to those of anti-Müllerian hormone (AMH), a well-established endocrine marker of the ovarian population of small antral follicles in cows. In the ovaries of 3-month calves, the population of 3 to 5 mm follicles contained the highest intra-follicular AMH amounts, and the number of 3 to 5 mm follicles on ovaries was closely correlated with AMH concentrations in the plasma of calves at this age (rs = 0.94). Before 3 months of age, only two out of six calves showed a clear postnatal FSH peak in plasma, and no correlation was found between plasma FSH and AMH concentrations. These results indicate that female calves undergo different patterns of FSH secretion and that postnatal activation of follicular growth up to the small antral stage appears independent and not directly related to circulating FSH levels.

Age-Specific Peripheral Anti-Mullerian Hormone (AMH) Concentration: A Candidate Endocrine Marker for Fertility Assessment in Cattle

Anti- Mullerian hormone (AMH) emerges as the most reliable endocrine marker in assessing the fertility potential over the ages. The age-specific reference range for peripheral AMH concentrations in cattle is lacking. The present study thus aimed (i) to address the question whether ovarian secretion of AMH would be affected by gonadotrophic status after gonadotrophin releasing hormone (GnRH) challenge in cattle and (ii) to establish age-specific plasma AMH concentrations in Holstein Friesian crossbred cattle (n= 151) using an enzyme immune-assay technique. Data on hormonal concentrations over time in GnRH-treated animals were analyzed by nonparametric one-way repeated measure ANOVA. GnRH challenge could not bring any change (P>0.05) in plasma AMH concentrations, while plasma FSH and LH concentrations changed significantly (P

Effects of genetic background on responses to superovulation in Japanese Black cattle.

We investigated the effects of genetic background on the responses to superovulation in Japanese Black cattle. The genotype frequencies of GRIA1 and FSHR relating to ovulation and follicular development in each of the major bloodlines—Tajiri, Fujiyoshi, and Kedaka—were analyzed. The Tajiri line had the lowest frequency of G allele homozygosity of c.710A>G in GRIA1 among the three bloodlines, and deviation from Hardy–Weinberg equilibrium was detected. Genotype frequencies of c.337C>G, c.871A>G, and c.1973C>G in FSHR were in Hardy–Weinberg equilibrium in all bloodlines. The results of generalized linear mixed-model analyses showed that farm, levels of plasma anti-Müllerian hormone (AMH) concentration, age in months, repeated superovulation, c.337C>G in FSHR, and bloodlines had significant effects on the responses to superovulation. The number of transferable embryos in the group heterozygous for c.337C>G in FSHR was significantly higher than that in the group homozygous for the C allele. The Kedaka line showed a significantly higher number of ova/embryos, fertilized embryos, and transferable embryos than the Tajiri and Fujiyoshi lines. The concentration of circulating AMH is a useful endocrine marker for antral follicle counts. This study revealed the effects of genetic background on the responses to superovulation using levels of plasma AMH concentration as a covariate. The prominent effect of genetic background on superovulation in the Kedaka line requires additional studies to confirm the genomic regions and polymorphisms that are involved in the trait.

Role of Anti-mullerian Hormone (AMH), As A Prediction of Clinical Pregnancy Outcome in Women with Polycystic Ovarian Syndrome with Sub Fertility

Background: Anti-mullerian hormone (AMH) in largely expressed throughout folliculogenisis (by granulosa cells in ovarian follicles) but highest in pre-antral and small antral steges (&lt;4mm) drameter of development.Objective: To analyse the usefulness of plasma anti-mullerian hormone (AMH) measurement as a test for assessing ovarian reserve in Polycystic ovary syndrome with sub fertility,Methodology: Women with PCOS and sub fertility, were included in this study with age group 18-35yrs. This Prospetive cohort study done in 30 cases in ShSMC with purposive randomized sampling, from January 2015 to December 2015. Serum, plasma &amp; for evaluation of PCOS, preferably TV-USG was done.Result: In this study we analyze 30 serum samples from patient aged between 18-35 years and transvaginal ultrasonography was done simultaneously. Mean age group of my patient was 25.2±4.7 years which correlates with AMH level in PCOS which is statistically significant (p&lt;0.001). Mean level of AMH is 5.7±4.9 mg/ml which is also correlate with transvaginal USG scan with the feature of PCOS (p&lt;.0.001). More than 50% woman presents with features of subfertility with PCOS and 46.7% are normal ovaries. Mean menstrual day 7.6 (4-12 days) which have a good linear correlation with irregularities of menstrual cycle and PCOS (p&lt;0.05).Conclusion: To conclude AMH appears to have a major inhibiting role during folliculogenesis, which may contribute to anovulation in PCOS. The reason for the raised AMH in PCOS may give clues as to the mechanism os anovulation.J Shaheed Suhrawardy Med Coll, June 2017, Vol.9(1); 3-5