Serum FSH Concentrations Research Articles

Effect of a GnRH agonist on the FSH receptors in prepubertal ewes

The present study was aimed at investigating the effects of the GnRH agonist, alarelin, on the expression of the follicle-stimulating hormone receptor (FSHR) in the pituitary gland. Also to evaluate the presence and immunolocalization of FSHR in the ovary, as well as to confirm the efficacy on the development of the follicles in the ovaries of ewes. Twenty-eight prepubertal ewes (5–6months of age) were randomly assigned to four experimental groups (EG, n=7 per group). The animals in experimental group EG-I, EG-II and EG-III were subcutaneously injected with 200μg, 300μg or 400μg alarelin antigens twice (day 0 and 14), respectively. Animals in the control group (CG) were twice subcutaneously injected with 2.0mL of a solvent (day 0 and 14). Samples of the pituitary gland and ovaries were collected aseptically on day 70 following treatment. Blood samples were taken from the jugular vein on day 0, 7, 14, 21, 28, 35, 50, 60 and 70 after the first alarelin antigen injection and ELISA used to measure the serum concentration of the GnRH antibody and FSH. Fluorescence quantitative RT-PCR was implemented to detect the gene expression of FSHR mRNA in the pituitary. Immunohistochemistry was performed to localize the FSHR in the ovary. GnRH antibody concentrations in the EG-I, EG-II and EG-III treatment groups increased gradually and were higher than that of the CG (P<0.05) or control from day 14 to day 60. Pituitary FSHR mRNA levels were significantly reduced (P<0.05) 1.38, 7.33 and 10.11 times in the EG-I, EG-II and EG-III groups, respectively. Immunohistochemistry identified that the immunostained cells of the FSHR were present in the ewes’ ovaries, which predominantly concentrated in the cytomembrane, cytoplasm and nuclei of the follicle cells. The oocytes had different immunostaining intensities at different developing stages. The gray values of microscopy images in EG-I, EG-II and EG-III groups increased, when compared to the CG. Ovaries in alarelin treated ewes of groups EG-II and EG-III were significantly higher (P<0.05), when compared to ewes in group EG-I and CG. The follicle vertical diameter (FVD), follicle transverse diameter (FTD), follicle-wall thickness (FWT), follicle externatheca thickness (FET) and follicle internatheca thickness (FIT) in the alarelin immunity ewes were greater than those in the CG. The serum FSH concentrations of the treated ewes remained higher than that in the CG (P<0.05). In conclusion, active immunity with alarelin stimulated the production of the GnRH antibody, inhibited the expression of FSHR mRNA in the pituitary gland, improved the distribution of FSHR in the ovary, increased the FSH secretion and thereby promoted the development of the ovaries and follicles in the ewes. This has important potential for developing a novel technique in the superovulation and regulation of reproductive functions in ewes.

Can the fall in serum FSH during coasting in IVF/ICSI predict clinical outcomes?

This retrospective cohort study determined whether the total falls in serum FSH and oestradiol concentrations from start to end of coasting in IVF/intracytoplasmic sperm injection could predict clinical outcomes. Ninety-nine cycles, with gonadotrophin-releasing hormone-agonist down-regulation where coasting with serial serum oestradiol and FSH monitoring was adopted due to risk of severe ovarian hyperstimulation syndrome, were consecutively included. The primary clinical outcome was live-birth rate (LBR); other outcomes measured were number of oocytes retrieved and fertilization, implantation and clinical pregnancy rates. LBR for FSH fall>10IU/l compared with 5–10 and<5IU/l were 45.4% versus 22.0% and 25.0%, respectively. Mean serum FSH fall was similar with and without live birth (8.4±6.2 versus 7.3±5.0IU/l) as were mean oestradiol and FSH concentrations on HCG administration, oestradiol fall, percentage fall in FSH/oestradiol and duration of coasting. None of the variables efficiently predicted live birth on regression analysis. The AUC of FSH fall was 0.53 at 11.0IU/l. Basal FSH, starting and total gonadotrophin dose and duration of coasting were positively correlated with FSH fall. A potentially clinically important association between live birth and FSH fall during coasting was apparent, which requires further evaluation.The purpose of this retrospective cohort study was to determine whether the magnitude of fall in the serum FSH and oestradiol concentrations from start to end of coasting in IVF/intracytoplasmic sperm injection cycles could predict the clinical outcomes. Gonadotrophin-releasing hormone-agonist down-regulated cycles (n=99), where coasting with serial serum oestradiol and FSH monitoring was adopted due to risk of ovarian hyperstimulation, were consecutively included. Live birth was the primary clinical outcome measured; number of oocytes retrieved and fertilization, implantation and clinical pregnancy rates were the other outcomes examined. Live-birth rate tended to be high when FSH fall was >10IU/l, compared with 5–10IU/l and <5IU/l, although not statistically significantly. Mean serum FSH fall were similar in live-birth and no-live-birth cycles (8.4±6.2 versus 7.3±5.0) as were mean oestradiol and FSH concentrations on hCG administration, oestradiol fall, percentage fall in FSH and oestradiol and duration of coasting. None of the variables efficiently predicted live birth. The area under the curve of FSH fall was 0.53. FSH fall of <11.0IU/l was found to be more likely to predict negative outcome (specificity 84.72%) than predicting positive outcome when FSH fall was >11IU/l (sensitivity 34.48%). Women’s basal FSH, starting and total gonadotrophin dose of ovarian stimulation and duration of coasting had direct positive correlation with the magnitude of FSH fall. A potentially clinically important rise in live birth in association with greater FSH fall during coasting was apparent, which requires further evaluation.

Successful pregnancy outcome following gamete intra-Fallopian transfer in a patient with Müllerian dysgenesis

A 29-year-old lady with Müllerian dysgenesis was keen to have a baby. Clinically, she was medium built with well-developed secondary female sexual characteristics. There was a short and blind vagina. She had undergone surgery for an imperforated hymen. Her FSH and LH concentrations were normal. Laparoscopy revealed a patent right Fallopian tube, a rudimentary right uterus and extensive pelvic endometriosis. She subsequently underwent gamete intra-Fallopian transfer (GIFT). Oocyte retrieval was carried out laparoscopically and a total of nine oocytes were retrieved. Four of the oocytes were transferred together with motile spermatozoa into the right Fallopian tube and the remaining five oocytes were inseminated with spermatozoa for IVF. Three embryos resulted and were frozen. She subsequently developed moderate ovarian hyperstimulation syndrome. Serum β-human chorionic gonadotrophin concentration 14days after GIFT was 1612IU/l. Her antenatal care was relatively uneventful until 31weeks of gestation when she was diagnosed to have intrauterine growth retardation and oligohydramnios. She then underwent an emergency Caesarean section at 32weeks of pregnancy delivering a normal baby.This case study describes a successful pregnancy outcome following gamete intra-Fallopian transfer (GIFT) in a woman with malformation of the vagina (Müllerian dysgenesis). A 29-year-old lady with Müllerian dysgenesis diagnosed at 16 years of age was keen to become pregnant. Upon examination, a decision was made for a William’s vulvovaginoplasty but as the patient was indecisive the surgery was deferred. Clinically, she is a medium-built lady with well-developed secondary female sexual characteristics. There was a short and blind vagina. Her serum FSH and LH concentrations were normal. Laparoscopy revealed a patent right Fallopian tube, a rudimentary right uterus and extensive pelvic endometriosis. She subsequently underwent GIFT. Nine oocytes were retrieved through laparoscopy. Four of the oocytes were transferred together with motile sperm into the right Fallopian tube and the remaining five oocytes were inseminated with sperm for IVF. Three embryos resulted and were frozen. Serum β human chorionic gonadotrophin concentration measured 14 days after GIFT was 1612IU/l. An abdominal ultrasonography performed at 5 weeks showed one intrauterine gestational sac. Her antenatal care was uneventful until 31 weeks of gestation when she developed a deficiency of amniotic fluid in the amniotic sac. She then underwent an emergency Caesarean section at 32 weeks of pregnancy. She delivered a healthy, normal 1.24kg baby boy. Her post-natal care was uneventful.

Chronic hyponatremia exacerbates multiple manifestations of senescence in male rats

The syndrome of inappropriate antidiuretic hormone secretion (SIADH) is frequently responsible for chronic hyponatremia in the elderly due to age-related disruption of the inhibitory component of brain osmoregulatory mechanisms. Recent research has indicated that chronic hyponatremia is associated with gait disturbances, increased falls, and bone fragility in humans, and we have found that chronic hyponatremia causes increased bone resorption and reduced bone mineral density in young rats. In this study, we used a model of SIADH to study multi-organ consequences of chronic hyponatremia in aged rats. Sustained hyponatremia for 18 weeks caused progressive reduction of bone mineral density by DXA and decreased bone ash calcium, phosphate and sodium contents at the tibia and lumbar vertebrae. Administration of 10-fold higher vitamin D during the last 8 weeks of the study compensated for the reduction in bone formation and halted bone loss. Hyponatremic rats developed hypogonadism, as indicated by slightly lower serum testosterone and higher serum FSH and LH concentrations, markedly decreased testicular weight, and abnormal testicular histology. Aged hyponatremic rats also manifested decreased body fat, skeletal muscle sarcopenia by densitometry, and cardiomyopathy manifested as increased heart weight and perivascular and interstitial fibrosis by histology. These findings are consistent with recent results in cultured osteoclastic cells, indicating that low extracellular sodium concentrations increased oxidative stress, thereby potentially exacerbating multiple manifestations of senescence. Future prospective studies in patients with SIADH may indicate whether these multi-organ age-related comorbidities may potentially contribute to the observed increased incidence of fractures and mortality in this population.

Co-administration of Mixed Steroid Hormones can Enhance the Recovery of Spermatogenesis Damaged by Gossypol Acetic Acid in Adult Rats

To determine whether co-administration of mixed steriod hormones can enhance the restoration of spermatogenesis damaged by gossypol acetic acid (GA). Adult male Wistar rats were treated daily for 8 weeks with GA at 50mg/kg plus testosterone undercanoate (100 mg/kg)/desogestrel (0.125 mg/kg)/mini-dose ethinylestradiol (0.025 mg/kg) (TU/DSG/EE), followed by a period of 9 weeks for recovery. Control animals were administered the same dose of GA or TU/DSG/EE, and vehicle, respectively. Testis weight, testicular sperm head count and histological analysis were utilized to assess the spermatogenesis. At the end of the 9-week reovery period, in rats given GA alone, spermatogenesis steadily declined. However, when rats received combined hormone administration during GA treatment, this decline was prevented and an complete recovery of spermagenesis occurred. The haploid spermatids and spermatocytes was not to be protected but to be more aggravatedly damaged. The excellent recovery must have resulted from that the hormone treatment could protect the ability of stem spermatogonia to differentialte and evolve progressively into mature spermatozoa. In addition, the concentrations of serum LH, FSH and intratesticular testosteron (ITT) notably decreased after 2 or 8 weeks of treatment, then returned to control levels at the end of 9-week recovery period. Steriod hormone treatment enhaces the recovery of spermatogenesis through preventing seminiferous epithelim from GA-induced destructive damage in rats. The enhanced recovery was closely associated with the marked suppression in intratesticular testosteron (ITT).

GnRH agonist active immunization influences gonadotropin receptor expression in pituitary gland, uterine development and secretion of peripheral reproduction hormones in female mice

Abstract Objective The present study was conducted to assess the effects of the GnRH agonist (alarelin) on the expression of GnRHR, FSHR and LHR mRNAs in the pituitary gland and uterine development in female mice. Methods Sixty pre-estrus female mice were randomly assigned to 4 groups (n = 15). The mice in experiment group 1 (EG-1), experiment group 2 (EG-2), and experiment group 3 (EG-3) were subcutaneously injected with 0.375 μg/kg, 0.75 μg/kg and 1.5 μg/kg alarelin antigen emulsion, respectively, once a day during 7 consecutive days to enhance immune response. Mice in the control group (CG) were injected with 0.l ml solvent once a day on 7 consecutive days. The samples of blood, pituitary and uterine horns were collected aseptically at day 21 from each mouse. Fluorescence quantitative RT-PCR was implemented to detect gene expression of GnRHR, FSHR and LHR in the pituitary glands. ELISA was used to measure the serum LH and FSH concentrations. The uterine slices were observed under an optical microscope;...

Follicular and endocrine profiles associated with different GnRH-antagonist regimens: a randomized controlled trial

This trial assessed the impact of early initiation of gonadotrophin-releasing hormone (GnRH) antagonist on follicular and endocrine profiles compared with the fixed GnRH-antagonist protocol. Eighty-five oocyte donors were randomized to GnRH antagonist starting in the mid-luteal phase of the prestimulation cycle (degarelix-ML group), on stimulation day 1 (early follicular phase, degarelix-EF group) or day 6 (fixed protocol) (mid-follicular phase, ganirelix-MF group). Subjects in the degarelix-EF and ganirelix-MF groups received placebo in the prestimulation cycle. At start of stimulation, serum concentrations of FSH (4.6 ± 2.3 versus 6.0 ± 1.8 IU/l), LH (2.7 ± 1.4 versus 4.7 ± 1.9 IU/l) and oestradiol (87 ± 35 versus 129 ± 50 pmol/l) were markedly lower ( P < 0.001) in the degarelix-ML group than in the placebo group. The coefficients of variation of follicle size (36.7 ± 5.5% versus 39.2 ± 9.4%) were not significantly different. No differences in endometrial histology, embryo quality and pregnancy rates in recipient cycles were observed between the regimens. In conclusion, early administration of GnRH antagonist altered the endocrine profile without modifying the follicular synchrony for the majority of subjects. Whether patients with a more heterogeneous follicle size at start of stimulation may benefit from an earlier intervention remains to be proven. The use of gonadotrophin-releasing hormone (GnRH) antagonist protocols in patients undergoing IVF, in which the antagonist is administered from stimulation day 5 or 6 (fixed protocol), has been associated with more heterogeneous follicle growth. Also, the hormone environment at start of stimulation in the GnRH-antagonist cycle may influence clinical response and outcome. The present trial assessed the impact of earlier start of GnRH-antagonist treatment on follicular and hormone profiles compared with the fixed protocol. Eighty-five oocyte donors were randomized to GnRH antagonist starting in the mid-luteal phase of the menstrual cycle prior to stimulation, on stimulation day 1 or on stimulation day 6 (fixed protocol). Subjects starting GnRH antagonist treatment on stimulation days 1 or 6 received placebo in the prestimulation cycle. At start of stimulation, serum concentrations of the hormones FSH, LH and oestradiol were markedly lower in the GnRH-antagonist group than in the placebo group. The follicle size heterogeneity was not different between the two groups. No differences in uterine histology, embryo quality and pregnancy rates in recipient cycles were observed between the regimens. The results indicate that the general IVF population will not benefit from an early start of GnRH-antagonist treatment. Whether patients with a poor response in a previous cycle, possibly due to a more heterogeneous follicle size at the start of stimulation, may benefit from an earlier start remains to be proven.

Reproductive function in men affected by X-linked adrenoleukodystrophy/adrenomyeloneuropathy

X-linked adrenoleukodystrophy (X-ALD) is the most frequent, severely neurodegenerative, clinically heterogeneous peroxisomal disorder, the signs of which are a consequence of myelin, adrenal cortex, and testes impairment. We studied testosterone, LH, and FSH levels in X-ALD/adrenomyeloneuropathy (AMN) patients. We evaluate the ability to procreate of these patients by analysis of pedigree and family screening by detection of very long-chain fatty acid (VLCFA) levels. Seventeen patients with X-ALD/AMN (16 with AMN and one asymptomatic) aged 24-48 (mean±S.D., 34.7±5.9) years, were identified based on the clinical picture, magnetic resonance imaging, and the presence of increased serum VLCFA levels. Nine X-ALD/AMN patients' daughters, mean ages ±S.D.=7.7±3.8 years, were identified as heterozygote by elevated VLCFA levels. Serum VLCFA levels were determined as ester derivatives by a gas chromatography method. Serum testosterone, LH, and FSH levels in X-ALD/AMN patients were detected by IRMAs. Serum testosterone levels were at the lowest levels of normal range but serum LH and FSH concentrations were increased in 57.1 and in 42.9% of X-ALD/AMN patients respectively. Among the 11 investigated of X-ALD/AMN married adult men, nine had produced offspring, a total of 13 children. All patients' daughters showed elevated serum VLCFA at heterozygote levels. In this study, we report that in a group of X-ALD/AMN married adult men, we did not find a significant decrease in fertility compared with the Polish population (18.2 vs 15%).

Triptorelin every other day can prevent premature LH surge: a strategy to reduce the daily gonadotrophin-releasing hormone agonists injections

The aim of the study was to compare the use of 100 mcg triptorelin every other day with the usual daily dose of leuprolide in terms of premature LH surge prevention and in vitro fertilization (IVF) outcomes. It was a prospective randomized study. Between January 2010 and December 2010, we selected 90 women with age between 18 and 40 years-old, that would be submitted to IVF cycle. Women with an elevated serum FSH concentration on cycle day 3 (FSH>10 IU/l) were excluded. A computer-based randomization was used to allocate the patients in two groups. In group 1, the pituitary desensibilization was performed with daily subcutaneous administration of 0,5 mg leuprolide acetate beginning in day 21 of the previous menstrual cycle. This dose was continued until ovarian quiescence was demonstrated by vaginal ultrasound following menstruation, when the dose was changed to 0,25 mg / day. In group 2, they received every other day 0,1 mg triptorelin acetate also beginning in day 21 of the previous menstrual cycle and keeping during all ovarian stimulation. All of them received the same protocol for ovarian stimulation using hMG according to the age and ovary response. The groups were compared according to the age, number of oocytes, number of mature oocytes, number of embryos, clinical pregnancy rate and occurrence of premature LH surge. Rates were expressed as percentage. The other results as means. The comparison of means were performed using t student test. To compare rates, it was used chi-square test. The groups didn't present difference in terms of patients' age, number of oocytes, number of mature oocytes, number of embryos and clinical pregnancy rate. Premature LH surge didn't occur in any groups. The use of triptorelin in every other day prevents premature LH surge with the same outcomes of daily leuprolide use. The advantage is a fewer number of injections the patients have to receive.

Diminished ovarian reserve is not independently associated with clinical reproductive loss in pregnancies achieved by assisted reproductive technologies (ART)

ObjectivePrior studies have suggested that women with diminished ovarian reserve (DOR) are at higher risk for reproductive loss, independent of age. These studies, however, did not distinguish very early losses from losses after clinical detection of an intrauterine pregnancy (IUP). Here we examined the rate of reproductive loss after sonographic identification of an IUP in a cohort of women with DOR compared to women with normal ovarian reserve undergoing ART.DesignRetrospective cohort.Materials and MethodsThe study cohort consisted of all fresh autologous ART cycles that resulted in a first singleton clinical IUP (sonographic identification of a single gestational sac at approximately 6-7 weeks gestational age) from Jan-Dec 2009 at a single fertility practice, verified by chart review. DOR was defined by basal serum FSH ≥14 IU/L. Patients were excluded if no FSH level was measured within one year of the cycle analyzed or if the patient had only one ovary. Patients were grouped by FSH <8, 8-13.9, or ≥14 IU/L.ResultsTabled 1FSH (IU/L)<88-13.9≥14# OF SUBJECTS60522125AGE (YEARS)33.535.035.6CLINICAL PREGNANCY LOSS24.3%28%28.% Open table in a new tab ConclusionAfter adjusting for age, there was no relationship between basal serum FSH concentration and pregnancy loss greater than 6-7 weeks gestational age in women achieving IUP through ART. The higher pregnancy loss rates previously reported in women with DOR may be due to loss prior to 6-7 weeks gestational age. ObjectivePrior studies have suggested that women with diminished ovarian reserve (DOR) are at higher risk for reproductive loss, independent of age. These studies, however, did not distinguish very early losses from losses after clinical detection of an intrauterine pregnancy (IUP). Here we examined the rate of reproductive loss after sonographic identification of an IUP in a cohort of women with DOR compared to women with normal ovarian reserve undergoing ART. Prior studies have suggested that women with diminished ovarian reserve (DOR) are at higher risk for reproductive loss, independent of age. These studies, however, did not distinguish very early losses from losses after clinical detection of an intrauterine pregnancy (IUP). Here we examined the rate of reproductive loss after sonographic identification of an IUP in a cohort of women with DOR compared to women with normal ovarian reserve undergoing ART. DesignRetrospective cohort. Retrospective cohort. Materials and MethodsThe study cohort consisted of all fresh autologous ART cycles that resulted in a first singleton clinical IUP (sonographic identification of a single gestational sac at approximately 6-7 weeks gestational age) from Jan-Dec 2009 at a single fertility practice, verified by chart review. DOR was defined by basal serum FSH ≥14 IU/L. Patients were excluded if no FSH level was measured within one year of the cycle analyzed or if the patient had only one ovary. Patients were grouped by FSH <8, 8-13.9, or ≥14 IU/L. The study cohort consisted of all fresh autologous ART cycles that resulted in a first singleton clinical IUP (sonographic identification of a single gestational sac at approximately 6-7 weeks gestational age) from Jan-Dec 2009 at a single fertility practice, verified by chart review. DOR was defined by basal serum FSH ≥14 IU/L. Patients were excluded if no FSH level was measured within one year of the cycle analyzed or if the patient had only one ovary. Patients were grouped by FSH <8, 8-13.9, or ≥14 IU/L. ResultsTabled 1FSH (IU/L)<88-13.9≥14# OF SUBJECTS60522125AGE (YEARS)33.535.035.6CLINICAL PREGNANCY LOSS24.3%28%28.% Open table in a new tab ConclusionAfter adjusting for age, there was no relationship between basal serum FSH concentration and pregnancy loss greater than 6-7 weeks gestational age in women achieving IUP through ART. The higher pregnancy loss rates previously reported in women with DOR may be due to loss prior to 6-7 weeks gestational age. After adjusting for age, there was no relationship between basal serum FSH concentration and pregnancy loss greater than 6-7 weeks gestational age in women achieving IUP through ART. The higher pregnancy loss rates previously reported in women with DOR may be due to loss prior to 6-7 weeks gestational age.

Exogenous hCG activity, but not endogenous LH activity, is positively associated with live birth rates in anovulatory infertility

Objective. To evaluate, retrospectively, the roles of endogenous and exogenous luteinising hormone (LH) activity on live birth rate in ovulation induction cycles.Methods. Associations between LH activity at baseline, end of stimulation and live birth rate were analysed in relation to patient characteristics, baseline and end of stimulation variables in WHO group II anovulatory women (n = 155) stimulated with recombinant follicle-stimulating hormone (rFSH) or highly purified human menopausal gonadotrophin (HP-hMG). HP-hMG provides FSH and exogenous LH activity mainly in the form of human chorionic gonadotrophin (hCG).Results. Serum LH concentrations at baseline or end of stimulation were not predictive of live birth rate in the rFSH group (n = 79) or HP-hMG group (n = 76). Serum hCG concentration at end of stimulation was a significant positive predictor in HP-hMG-treated women. Other variables were not independently predictive of live birth in either of the groups, except for a negative association between serum FSH concentrations at the start of stimulation and live birth in the rFSH-treated group.Conclusions. Endogenous LH concentrations are not predictive of live birth in anovulatory WHO group II patients undergoing ovulation induction with rFSH or HP-hMG. On the other hand, exogenous hCG activity during HP-hMG stimulation is positively associated with treatment outcome.

The Release of LH in Response to Kiss-10 Is Dependent to the Age and Influenced by Serum Concentrations of Estradiol (E2) in Heifers in Tropical Regions.

The objective of the study was to evaluate the influence of the age of heifers on the release of LH in response to Kiss-10. Heifers (4, 7, and 11 months old) received an i.v. injection of kiss-10 (5 micrograms/kg). Serum concentrations of LH, FSH and estradiol (E2) were quantified by radioimmunoassay and data were subjected to analysis of variance. Serum concentrations of LH and FSH were analyzed in a repeated measurements design in blood samples collected every 15 minutes from 4 hours before to 4 hours after administration of Kiss-10. Serum concentrations of E2 were analyzed in a completely randomized design from a blood sample collected just before treatment with Kiss-10. Serum concentrations of FSH were similar (P > 0.3) in heifers of 4, 7, and 11 months old (21 ± 2.2, 18 ± 2.2, and 15 ± 2.4 ng/ml, respectively) and administration of Kiss-10 increased (P < 0.01) secretion of FSH compared to the concentrations during the pre-treatment period (33 ± 1.9 vs. 18 ± 1.3 ng/ml), and the magnitude of the stimulatory effect of Kiss-10 was similar among age groups. Similarly, serum concentrations of LH did not differ (P > 0.1) in heifers of 4, 7, and 11 months old (1.4 ± 0.26, 2.3 ± 0.41, and 2.3 ± 0.4 ng/ml, respectively). In all three age groups LH increased (P < 0.01) in response to Kiss-10 compared to the pre-treatment period (3.7 ± 0.60 vs. 1.1 ± 0.21 ng/ml). However, the stimulatory effect of Kiss-10 on LH secretion was higher (P < 0.01) in the 11 versus the 4 month old heifers when evaluated as either mean LH (4.8 ± 0.88 vs. 2.6 ± 0.41 ng/ml) or area under the curve of LH (36.5 ± 7 vs. 18.9 ± 3 arbitrary units). The greater LH secretion induced by Kiss-10 detected in 11 month old heifers was associated with higher (P < 0.06) serum concentrations of E2 compared to 4 month old heifers (1.68 ± 0.51 vs. 0.43 ± 0.11 pg/ml). We speculate that increased ovarian activity that occurs as the heifers approach puberty increases sensitivity of the hypothalamic-pituitary axis to Kiss-10. (poster)

Sequential use of letrozole and gonadotrophin in women with poor ovarian reserve: a randomized controlled trial

Sequential use of letrozole and human menopausal gonadotrophin (HMG) was compared with HMG only in poor ovarian responders undergoing IVF. Patients ( n = 53) with less than four oocytes retrieved in previous IVF cycles or less than five antral follicles were randomized to either letrozole for 5 days followed by HMG or HMG alone. The letrozole group had lower dosage of HMG ( P < 0.001), shorter duration of HMG ( P < 0.001) and fewer oocytes ( P = 0.001) when compared with controls. Live-birth rate was comparable with a lower miscarriage rate in the letrozole group ( P = 0.038). Serum FSH concentrations were comparable in both groups except on day 8, while oestradiol concentrations were all lower in the letrozole group from day 4 (all P < 0.001). Follicular fluid concentrations of testosterone, androstenedione, FSH and anti-Müllerian hormone were higher in the letrozole group ( P = 0.009, P = 0.001, P = 0.046 and P = 0.034, respectively). Compared with HMG alone, sequential use of letrozole and HMG in poor responders resulted in significantly lower total dosage and shorter duration of HMG, a comparable live-birth rate, a significantly lower miscarriage rate and a more favourable hormonal environment of follicular fluid. The management of poor ovarian responders or women with poor ovarian reserve in IVF is controversial. The use of letrozole has been studied; however, results are inconsistent. This randomized trial studied the sequential use of letrozole and gonadotrophin compared with gonadotrophin alone in poor responders undergoing IVF. The sequential use of letrozole and gonadotrophin led to a significantly lower dosage and shorter duration of gonadotrophin use, significantly fewer oocytes, comparable live-birth rate, a significantly lower miscarriage rate and a more favourable hormonal environment at a lower cost.

Exercise affects both ovarian follicular dynamics and hormone concentrations in mares

The objectives were to evaluate the effects of exercise on ovarian folliculogenesis and related hormones in mares. Mares (n = 11) were randomly assigned into a control (non-exercised) or treatment (exercised) group. Treatment mares (n = 5) were moderately exercised for 30 min, 6 d/wk. All mares underwent daily transrectal ultrasonographic examinations and ovarian follicles > 6 mm were measured. Blood samples were collected during the first (Cycle 1) and last (Cycle 4) cycle, and serum concentrations of cortisol, LH, and FSH were determined. Mean cortisol concentrations were elevated (P < 0.05) in exercised mares, 6.29 ± 0.22 compared with 5.62 ± 0.16 ng/dL (mean ± SEM), 30 min post exercise. There were no significant differences between groups in mean FSH concentrations; however, exercised mares had lower (17.3 ± 6.4 vs 41.1 ± 5.5 ng/mL; P < 0.05) peak LH concentrations. Furthermore, exercised mares experienced a longer (24.7 ± 0.8 vs 22.2 ± 0.8 d; P < 0.05) mean interovulatory interval for all cycles combined, fewer (P < 0.05) follicles 6 to 20 mm in diameter, and an increased (P < 0.05) number of follicles >20 mm following deviation. The dominant and largest subordinate follicle in exercised mares had a greater (P < 0.05) mean diameter on the day of deviation, suggesting delayed deviation. Exercised mares also tended (P = 0.06) to have an increased number of cycles with at least two dominant follicles compared to control (62 vs 36%, respectively), indicating a decreased ability of the largest follicle to assert dominance. Under the conditions of this study, moderately exercising mares induced higher cortisol concentrations, lowered peak LH concentrations, and altered ovarian follicular dynamics.

GnRH agonist active immunization influences ovarian development and GnRH receptor mRNA expression levels of pituitary in Japanese white rabbits ( Oryctolagus cuniculus)

The present study was conducted to assess the effects of active immunization against the gonadotropin-releasing hormone agonist (GnRHa) on ovary development and GnRH receptor mRNA expression levels in pituitary in Japanese white rabbits ( Oryctolagus cuniculus). Twenty-four three-month-old rabbits were randomly divided into four groups (n = 6). The animals were subcutaneously injected with 100 μg of GnRHa (alarelin) antigen in experimental group I (EG-I), 100 μg in experimental group II (EG-II) and 50 μg in experimental group III (EG-III) respectively. Alarelin antigens were re-injected in EG-II and EG-III with the same dosage on 20 days. CG was blank. The samples of ovaries and uteri were collected aseptically at the end of the experiment (70 days). Tissue slices were observed under light and electron microscopes. Serum anti-GnRH antibody titers and concentrations of FSH and LH were measured with ELISA. The anti-GnRH antibody titers reached the peak levels respectively at 30 days in EG-I (1:800), at 40–50 days in EG-II (1:1600) and at 40–50 days in EG-III (1:800). From 40 to 70 days, anti-GnRH antibody titers in EG-II were higher than that in EG-I and EG-III ( p < 0.05). The highest FSH levels in EG-II and EG-III were detected on 40 days. FSH level in EG-II was higher than those in EG-I, CG ( p < 0.01) and EG-III ( p < 0.05) on 40 days. There was no significant difference in FSH levels between EG-I and CG. Serum LH concentrations in EG-II and EG-III reached the peak levels on 50 days and 40 days, LH level in EG-II exceeded other 3 groups on 50 days ( p < 0.05). Real-time quantitative PCR analysis indicated that the levels of FSH-β mRNA and GnRH-R mRNA in pituitary significantly declined, while LH-β mRNA increased at 70 days following alarelin antigen treatment. The results also showed that alarelin antigen treatment improves the development of ovaries, follicles and uteri in Japanese white rabbits. Alarelin antigen treatment and re-injection with 100 μg dose had better effects than 50 μg treatment. This may have significance for the development of novel GnRH-based techniques in regulative functions in rabbits.

FSH receptor gene polymorphisms in fertile and infertile Han-Chinese males

BackgroundFollicle stimulating hormone receptor (FSHR), which mediates the effects of FSH, is essential for normal spermatogenesis and male reproduction. This study aimed to investigate the effects of the FSHR polymorphisms on idiopathic male infertility and serum FSH levels in Han-Chinese population. MethodsA case–control study was conducted with 364 idiopathic infertile patients (97 nonobstructive azoospermic, 79 oligozoospermic and 188 normozoospermic) and 281 fertile controls. Three polymorphisms at nucleotide position −29 and codons 307 and 680 of the FSHR gene were genotyped by Taqman allelic discrimination and RFLP. FSH levels were measured by RIA. ResultsThe allele and genotype frequencies of these three polymorphisms were not significantly different between each group of the cases and controls. Serum FSH concentrations did not differ between subjects with different genotypes within each group. Together the three SNPs mainly formed four discrete haplotypes. The distribution of the haplotype was not different between each group of infertile men and fertile controls and did not influence serum FSH levels in each group. ConclusionsOur findings suggest that the FSHR polymorphisms at the studied sites do not associate with idiopathic male infertility and have no influence on FSH levels both in normal and infertile males in the Han-Chinese population.

Limited Value of Ovarian Function Markers following Orthotopic Transplantation of Ovarian Tissue after Gonadotoxic Treatment

In young women, some treatments for cancer or other conditions (such as sickle cell anemia) may give rise to primary ovarian insufficiency. Ovarian transplantation is one of the available options for fertility preservation, with highly variable pregnancy rates. The objective of the study was to investigate markers of ovarian reserve and ovarian function in women up to 7 yr after orthotopic ovarian transplantation. Secondary objectives were to assess the relationship between markers of ovarian reserve and pregnancy rate along with the duration of ovarian function. This was a prospective cohort study in 10 women, with a mean follow-up of 2.5 yr. The study was conducted at a university hospital in Brussels, Belgium. Patients included 10 women who were about to receive or had previously received gonadotoxic treatment. In seven women cryopreservation of ovarian tissue was performed before starting treatment. Subsequently autografts were orthotopically transplanted in these women. Three women, who had already developed primary ovarian insufficiency due to treatment, underwent orthotopic transplantation of ovarian allograft tissue originating from their human leukocyte antigen-compatible sisters. Serum concentrations of FSH, LH, estradiol, inhibin B, and anti-Müllerian hormone (AMH) were measured. On average, first menses took place after 4.7 months. Duration of graft functioning varied from 2 to more than 60 months. FSH concentrations remained elevated, whereas estradiol levels normalized and AMH was low to undetectable. Inhibin B varied among women and was not associated with the duration of ovarian function (hazard ratio 0.966, 95% confidence interval 0.881-1.059). Two spontaneous pregnancies occurred. Endocrine characteristics were not significantly different in these women. Low AMH and inhibin B concentrations may suggest decreased ovarian reserve in women after ovarian transplantation. AMH and inhibin B levels may not be associated with the duration of ovarian graft function or probability to achieve a pregnancy.

Somatostatin 14 affects the pituitary-ovarian axis in infant rats

The effects of multiple somatostatin (SRIH-14) treatment on the pituitary-ovarian axis were examined in infant rats. Female Wistar rats received subcutaneously two daily 20 μg/100g b.w. doses for five consecutive days (from 11 to 15 days of age). Changes in cell volume, volume density and number per unit area (mm²) of follicle-stimulating (FSH), luteinizing (LH) and somatotropic (GH) immunolabeled cells were evaluated by stereology and morphometry. Serum FSH and LH concentrations were determined by RIA. Ovaries were analyzed by simple point counting of follicles. SRIH-14 treatment significantly reduced FSH and LH cell volume, while their volume density and number per unit area were unaltered. Serum concentrations of FSH and LH were significantly reduced. Volume and volume density of GH cells was significantly decreased after SRIH-14 treatment, while their number per unit area was unaltered. In the ovary, SRIH-14 induced a significant increase in the percentage of primordial follicles followed by a significant decrease in percentage of primary follicles. The number of healthy and atretic preantral follicles was unchanged. It can be concluded that SRIH-14 treatment during the infantile period markedly inhibits pituitary FSH, LH and GH cells. In the ovary, SRIH-14 acts by inhibiting initial folliculogenesis without affecting atretic processes.

Frequency distribution of polymorphisms in the FSH receptor gene in infertility patients of different ethnicity

Studies on the frequency distribution of follicle-stimulating hormone receptor (FSHR) polymorphisms report conflicting results. It has been suggested that ethnicity might influence these outcomes. Therefore, the aim of this study was to determine the frequency distribution of FSHR polymorphisms at position 680 of exon 10 within a large group of women with fertility problems from different ethnic backgrounds. A total of 1771 women of different ethnic origin (Caucasian, Asian, Hindustani, Creole and Mediterranean) were studied. FSHR single-nucleotide polymorphisms at codon 680 of exon 10 were determined by restriction fragment length polymorphism of amplicons generated by polymerase chain reaction. Genotypes were compared with serum FSH concentrations and between different ethnic groups. A significantly lower number of Asians (10.5%) were found to have the Ser680Ser receptor variant compared with Caucasians (21.5%) and Mediterraneans (22.3%) ( P = 0.010). FSH concentrations did not differ between the various ethnic groups, or the different FSHR polymorphisms. In conclusion, the Ser680Ser receptor variant is less common in the Asian subgroup compared with Caucasians and Mediterraneans. This indicates that, when comparing allelic frequency distributions of the FSHR polymorphism variants, ethnic background should be accounted for. FSH concentrations did not differ between FSHR polymorphisms or between ethnic groups.

Progesterone as the driving regulatory force behind serum FSH concentrations and antral follicular development in cycling ewes

Ovarian antral follicles in sheep grow in an orderly succession, producing typically three to four follicular waves per 17-day oestrous cycle. Each wave is preceded by a transient increase in circulating FSH concentrations. The mechanism controlling the number of recurrent FSH peaks and emerging follicular waves remains unknown. During the ewe's oestrous cycle, the time between the first two FSH peaks and days of wave emergence is longer than the intervals separating the ensuing FSH peaks and follicular waves. The prolonged interpeak and interwave interval occurs early in the luteal phase when low levels of progesterone are secreted by developing, or not fully functional, corpora lutea (CL). The purpose of the present study was to determine the effect of varying progesterone (P(4)) levels on circulating concentrations of FSH and antral follicular development in sheep. Exogenous P(4) (15 mg per ewe, i.m.) was administered twice daily to six cycling Rideau Arcott×Dorset ewes from Day 0 (ovulation) to Day 4 (the mean duration of the interwave interval); six animals served as controls. Follicular growth was monitored in all animals by daily transrectal ultrasonography (Days 0-9). Jugular blood samples were drawn twice a day from Day 0 to Day 4 and then daily until Day 9 to measure systemic concentrations of P(4), FSH and 17β-oestradiol (E(2)). The first FSH peak after ovulation was detected on Days 1.5±0.2 and 4.2±0.2 in treated and control ewes, respectively (P<0.05). The next FSH peak(s) occurred on Day 3.9±0.3 in the treated group and on Day 6.4±0.5 in the control group. Consequently, the treated group had, on average, three follicular waves emerging on Days 0, 3 and 6, whereas the control group had two waves emerging on Days 0 and 5. Mean serum E(2) concentrations were greater (P<0.05) in control compared with treated ewes on Days 1.3, 2.3, 3.3, 4.0 and 4.3 after ovulation. In summary, creation of mid-luteal phase levels of P(4) in metoestrus shortened the time to the first post-ovulatory FSH peak in ewes, resulting in the emergence of one more follicular wave compared with control ewes during the same time frame. Therefore, P(4) appears to be a key endocrine signal governing the control of periodic increases in serum FSH concentrations and the number of follicular waves in cycling sheep.