Increase In LH Research Articles

OR32-3 Kisspeptin- a Novel Clinical Test of Hypothalamic Function in Men with Hypogonadotrophic Hypogonadism

Background: Hypogonadotrophic Hypogonadism is characterised by hypogonadism in the context of low / inappropriately normal gonadotrophin levels. Congenital HH (CHH) occurs due to defective hypothalamic GnRH neuronal migration (e.g. Kallman’s syndrome), or secretion. However, no direct test of hypothalamic GnRH neuronal function currently exists. Kisspeptin is a neuropeptide that stimulates endogenous hypothalamic GnRH release. Thus, we investigated whether kisspeptin could be used to interrogate hypothalamic function in men with CHH. Methods: Men with CHH (low testosterone, low LH/FSH, normal MRI pituitary / baseline pituitary function, absent puberty, unprimed by pulsatile GnRH; n=4) and healthy eugonadal men (n=20) received an intravenous bolus of GnRH (100mcg), or kisspeptin-54 (6.4nmol/kg), on two study visits ≥1 week apart. Serum gonadotrophins were measured every 15mins for 6hrs following injection. Increases in serum gonadotrophins from baseline following GnRH / kisspeptin in eugonadal men and CHH were compared by unpaired t test. Results: Mean increase in serum LH from baseline was 8.2±3.8iU/L in eugonadal men and 0.12±0.13iU/L in CHH (P=0.0003) following kisspeptin administration. All eugonadal men had an LH increase >1.5iU/L, whereas all men with CHH had an LH increase <1.5iU/L following kisspeptin. In contrast, mean increase in serum LH from baseline following GnRH was 6.2±3.2iU/L in eugonadal men and 2.2±3.8iU/L in CHH (P=0.062). Therefore, whilst the kisspeptin-induced LH increase effectively discriminated men with HH from eugonadal men (area under ROC 1.0), GnRH-induced LH increase was less discriminatory (area under ROC 0.82). In eugonadal men, the maximal increase in LH following kisspeptin significantly predicted the maximal increase in LH following GnRH (univariate linear regression, r2=0.45; P=0.0013), however this relationship was lost in men with HH (r2=0.03; P=0.83). Conclusion: Collectively, these data confirm that a novel kisspeptin test of hypothalamic GnRH function can better discriminate men with CHH from eugonadal men than currently available investigations (such as GnRH test). Therefore, a kisspeptin test could offer significant clinical benefit for the accurate diagnosis and management of patients with hypogonadism.

SAT-405 Activation of Kiss1 Neurons in the Posterodorsal Medial Amygdala Stimulates Gonadotropin Release via the Release of Kisspeptin in Female Mice

Reproduction is regulated by a complex neuronal network the precise components of which are still unknown. Kisspeptins, secreted from Kiss1 neurons in the arcuate nucleus (Kiss1ARC) and anteroventral periventricular/periventricular (Kiss1AVPV/PeN) nuclei, have been directly linked to the release of GnRH from the hypothalamus and hence, LH from the anterior pituitary. However, there is a population of Kiss1 neurons in the posterodorsal medial amygdala (Kiss1MePD) whose contribution to GnRH/LH stimulation has not yet been described. Here, we aimed to determine whether the release of kisspeptin and/or other components, besides kisspeptin, within the Kiss1MePD neuron, can stimulate LH release into the peripheral circulation. In this regard, we used a chemogenetic approach to specifically activate Kiss1MePD neurons in adult Kiss1Cre/+(heterozygous state) females which we studied in parallel to Kiss1Cre/Cre(Kiss1 knock-out state) littermates (n=5/group). Females received bilateral stereotaxic injections of an adeno-associated virus (pAAV) encoding a Cre-driven Gq-coupled excitatory DREADD (pAAV5/hSyn-DIO-hm3Dq:mCherry; titer 3x1012 genome copies per ml; 1 µl per hemisphere). Following infection, mice were given 3 weeks for recovery and maximum expression of the AAV vector. On day 1 of the experiment, animals were administered an ip bolus injection of vehicle saline (0.9% NaCl; day 1) and then hM3D receptors were activated by ip injection of its agonist, clozapine N-oxide (CNO; 10 mg/kg dissolved in saline; day 2). Blood samples were collected just before saline or CNO treatment (0) and then every 15 min for 90 min. Kiss1Cre/+mice expressing hM3Dq:mCherry in the MePD and treated with CNO to activate the Kiss1MePD neurons, showed an increase in LH within 30 min after the injection (P=0.0107) compared to animals receiving saline treatment, which was sustained for a further 30 min before returning to basal levels. Furthermore, no alteration in LH was observed in Kiss1Cre/Cre(i.e., Kiss1 KO) animals treated with either saline or CNO indicating that kisspeptin is the only component within the Kiss1MePD neuron that has the ability to stimulate LH release. Analysis following the completion of pharmacological studies demonstrated that mCherry expression was evident in 89% of the targeted Kiss1MePD neurons. Interestingly, mCherry labeled projections were observed in the ARC and specifically in close contact with Kiss1ARC neurons. Thus, in the female mouse, Kiss1MePD neurons have the ability to stimulate the gonadotropic axis but only when kisspeptin is present. Furthermore, this study suggests that this action is, at least in part, via the activation of Kiss1ARC neurons which, in turn, stimulate GnRH/LH release. Overall, our data provide insight into the potential mechanism via which amygdala regulated social cues can enhance reproductive function.

SAT-425 NKB Signaling in the Posterodorsal Medial Amygdala Stimulates Gonadotropin Release in a Kisspeptin-Independent Manner in Female Mice

Female reproduction is regulated by a complex signaling network, the function of which is dependent on the level of circulating estrogens. Neurokinin B (NKB) colocalizes with kisspeptin in Kiss1 neurons of the arcuate nucleus (ARC). The autosynaptic action of NKB via the neurokinin 3 receptor (NK3R) in these neurons induces kisspeptin release in the presence of sex steroids. This has led us to believe that NKB stimulates LH secretion in a kisspeptin-dependent manner despite the latter mechanism having been investigated in the persistent hypogonadal state [e.g. Kiss1r knock out (KO) mice and agonadal monkeys]. Here, we aimed to assess whether in the presence of estradiol, senktide, a NK3R specific agonist, can stimulate LH release in a kisspeptin-independent manner. Adult WT female mice were ovariectomized (OVX) or OVX and estradiol-treated (OVX+E2) and studied in parallel to Kiss1KO or Kiss1KO+E2 littermates (n=10/group). Intracerebroventricular (icv) injections of senktide (600pmol) were performed and blood samples were collected before and 25 min after icv injection. Interestingly, the presence of E2 in Kiss1KO females resulted in LH increase like that observed in WT animals. Subsequently, we aimed to locate the brain area which senktide may be acting to stimulate LH release in Kiss1KO+E2 mice. We mapped the distribution of NK3R in the mouse hypothalamus and moreover investigated the anatomical relationship of NK3R and GnRH expression with dual-label immunohistochemistry as a likely pathway of the kisspeptin-independent stimulation of LH release exerted by NKB. We observed no instances of co-expression between NK3R and GnRH cell bodies (>100 cells analyzed from a total of 16 mice) and only minimal close appositions between the two proteins in the ARC and medial septum. Thus, the kisspeptin-independent action of NKB cannot be attributed to direct stimulation of NK3R located on GnRH neurons. Interestingly, we observed that NK3R immunopositive cells in the posterodorsal medial amygdala (MePD), unlike the ARC, were upregulated in the presence of E2. Next, WT OVX+E2 and Kiss1KO+E2 females were injected with senktide (600pmol; n=5/group) in the ARC, the AVPV/PeN or the MePD; areas found to contain NK3R-expressing neurons, and blood samples were collected before and then every 15 min for 45 min. Senktide significantly stimulated LH secretion in all three areas in WT OVX+E2, but only in the MePD of Kiss1KO+E2 mice. Conversely, in the absence of E2, Kiss1KO females injected with senktide in the MePD did not show any alteration in LH, mimicking the results we obtained after icv administration. These data demonstrate that activation of NK3R in the amygdala stimulates LH release in a kisspeptin independent manner in the presence of estrogen. This novel pathway for LH release, that bypasses the kisspeptin neuron, may play an important role in linking amygdala regulated social cues with gonadotropin release.

SAT-222 Hormone Responses Associated with Letrozole Ovulation Induction in Anovulatory Women with PCOS

: The aromatase inhibitor (AI) letrozole has been shown to be the most effective oral medication for ovulation induction in anovulatory women with PCOS. It has been established that AIs block the conversion of androgens to estrogen, leading to reduction of serum estrogen levels. However, the mechanism by which letrozole induces ovulation is unknown. It has been suggested that the mechanism of AIs in ovulation induction may involve increased FSH secretion in response to reduced estrogen levels. To examine the mechanism by which AIs induce ovulation, we performed a pilot interventional study. Ten anovulatory women with PCOS, aged 24-34 years were enrolled. Each subject underwent baseline serum hormone assessment and transvaginal ultrasound. Women were excluded if they had hypogonadotropic hypogonadism, or ultrasound evidence of a dominant follicle or corpus luteum. Each subject underwent ovulation induction with letrozole 2.5 mg/day for 5 days. Repeat transvaginal ultrasound was done three days after completing letrozole. Ovulation was defined by (1) presence of >=1 ovarian follicle >=16 mm on follow up ultrasound and (2) positive home ovulation predictor test. Serum hormone levels were assessed in all subjects at baseline and at two, four, and seven days after letrozole initiation. Mean serum hormone levels were compared using paired Student’s t test with statistical significance set at p<0.05. Three of 10 subjects ovulated after completing letrozole. In all subjects, the baseline levels (mean ± SD) of serum FSH, LH, estradiol, and androstenedione were 5.4 ± 2.2 mIU/mL, 12.1 mIU/mL, 86.2 ± 32.1pg/mL, and 3.2 ± 1.5 ng/mL, respectively. A significant reduction in serum estradiol was noted on day two compared to baseline (-35.2 pg/mL). There was a significant increase in mean serum FSH (+1.8 mIU/mL) and LH (+4.7 mIU/mL) on day two after starting letrozole. Significant increases in serum androstenedione were observed on day two (+0.8 ng/mL) and day four (+1.5 ng/mL) relative to baseline values. The study was not sufficiently powered to detect differences in serum hormone levels between women that did and did not ovulate in response to letrozole administration. These data suggest that the mechanism of ovulation induction by AI may involve a transient increase in serum gonadotropins resulting from interrupted estrogen negative feedback. Sources of Research Support: NIH Grants T32 HD007203, K12 HD001259, and P50 HD012303.Key Words: Polycystic Ovary Syndrome; Infertility; Anovulation

SUN-485 Investigating the Metabolic and Reproductive Role of AR in Leptin Receptor Expressing Cells

Androgens are important for reproductive and metabolic physiology of males and females. Androgens can act upon androgen receptors (AR), which are expressed in many tissues. In the brain, AR is abundant in hypothalamic nuclei involved in the regulation of reproduction and metabolism. While previous studies have examined the contribution of estrogens or estrogen receptors, the role of androgens acting on AR on the central regulation of reproduction and metabolism is less well understood. We have developed an in situ hybridization probe to target AR mRNA in mice. Our preliminary studies have shown that AR is abundant in hypothalamic nuclei, including the arcuate and ventral premammillary nucleus, and is expressed in key neuronal populations associated with the neuroendocrine reproductive axis and the metabolic control of reproduction. One of these neuronal populations expresses the leptin receptor (LepR). The adipocyte secreted hormone leptin signals energy stores to the hypothalamic-pituitary-gonadal axis. Both humans and mice with lack of leptin or LepR are obese, remain in a pre-pubertal state, and are infertile. LepR are highly enriched in the hypothalamus, including areas associated with regulation of reproduction and metabolism. Our hypotheses are that AR expression in LepR cells is necessary for reproduction and metabolism, and that androgens acting on AR in LepR cells mediate the pathophysiology of PCOS. To test our hypothesis, we have generated a genetic mouse model to delete AR from LepR expressing cells (LepRARKO). Our findings show that pubertal timing, estrous cycles, fertility, body weight, and glucose homoestasis do not change in female LepRARKO mice. Male LepRARKO mice show no difference in pubertal timing, fertility, body weight or glucose homeostasis, yet show an increase in spontaneous ambulatory activity during the light phase. Since androgens provide negative feedback to the HPG axis, we investigated whether loss of AR from LepR neurons resulted in changes in gonadotropin levels with gonadectomy. Male LepRARKO mice that were orchidectomized (ORX) show an exaggerated increase in serum LH compared to ORX control males, while FSH and testosterone levels were not significantly different. Completion of our studies will further expand our knowledge of the role that androgens acting on AR play in reproductive and metabolic physiology of males and females.

SUN-221 Effects of Liraglutide on Obesity-Associated Functional Hypogonadism in Men: A Randomized, Testosterone Replacement Treatment-Controlled Study

Objective: Obesity causes functional hypogonadism(FH) due to suppression of hypothalamus-pituitary-testicular (HPT) axis that is potentially reversible. Weight reduction with lifestyle measures (LSM) should be recommended as a first line treatment approach. In clinical practice, LSM often fails and may be insufficient to relieve symptoms of FH. A role of testosterone replacement treatment (TRT), after a trail of unsuccessful LSM, is unclear. In selected patients, TRT could be started concomitantly or in addition to LSM to augment the benefits of LSM, although the quality of evidence supporting this concept is low. GLP-1 receptor agonist liraglutide is linked to progressive and sustained weight loss. Furthermore, a potential direct impact of GL-P1 agonism on hypothalamus-pituitary-testicular (HPT) axis was reported in preclinical models. Despite the fact, that the prevalence of FH in obese men is high, the impact of liraglutide on FH in obese men with or without diabetes has not yet been addressed in a randomized prospective clinical study. Aim: We aimed to compare the effects of liraglutide and TRT on FH in obese men that had been poor responders to LSM, by means of reversal of FH and weight reduction. Design:We designed 16-week prospective randomized open-label studywith 30 men (aged 46.5±10.9years, BMI 41.2±8.4 kg/m2, mean ± SD) that were randomized to liraglutide 3.0 mg QD (LIRA) or 50 mg of 1% transdermal gel QD (TRT). Methods: Sexual function with standardized questionnaires and anthropometric measures were assessed. A fasting blood was drawn for determination of endocrine and metabolic parameters followed by OGTT. Model-derived parameters including HOMAIRand calculated free testosterone (cFT) were calculated.Results: Total testosterone (+5.9±7.2 in TRT vs +2.6±3.5 nmol/l in LIRA) and the sexual function significantly increased in both arms, with no significant between treatment differences. SHBG tended to increase in LIRA. There was a significant differential effect on HPT axis resulting in further suppression of LH and FSH in TRT and a significant increase of LH and FSH in LIRA (p<0.001). Subjects treated with LIRA lost on average 7.9±3.8 kg compared with a 0.9±4.5 kg loss in TRT (P<0.001). LIRA was also superior in reduction of waist circumference. Metabolic syndrome was resolved in two patients in LIRA and in no subjects in TRT.Conclusions: When LSM fails and bariatric surgery is not yet indicated, anti-obesity treatment with liraglutide should be advised over TRT, by means of overall health benefit improvement in men with obesity-associated FH. Future studies should investigate the complex crosstalk between GLP-1 and HPT axis beyond the mere impact of the magnitude of weight reduction.

Role of GnRH in the ontogeny and regulation of the fetal hypothalamo-pituitary-gonadal axis in sheep

Adult reproductive ability is to a large extent determined by the appropriate development of the reproductive axis during fetal life. Studies have investigated the role of the fetal hypothalamus in the ontogeny and regulation of pituitary gonadal function during fetal development in sheep. Using immunocytochemistry, we examined the ontogeny of gonadotroph development in the pituitary of female sheep fetuses. At day 70 of gestation (term = 145 days), only immunopositive LH beta cells were present. The number and intensity of staining of these LH beta cells had increased by day 100 but had declined again by day 130. Immunopositive alpha-subunit and FSH beta cells appeared at day 100 of gestation and had further increased in number and staining intensity by day 130 of gestation. Treatment of fetuses with the GnRH agonist buserelin resulted in desensitization of the fetal pituitary gonadotrophs, inhibition of pituitary LH beta and FSH beta mRNA expression and a reduction in the number of immunopositive gonadotrophin-containing cells. Pulsatile GnRH treatment resulted in pituitary-gonadal activation and an increase in LH, FSH and testosterone secretion in males. Thus, the synthesis and secretion of the gonadotrophins during fetal development is critically dependent on the secretion of GnRH from the fetal hypothalamus. Inhibition of fetal gonadotrophins by buserelin treatment from day 70 of gestation resulted in a 40% reduction in the size of the fetal testis at birth, and there were no effects on the fetal ovaries. This reduction in testis size was due to a 45% reduction in the number of Sertoli cells. However, when buserelin was given between day 70 and day 110 of gestation, there were no effects on testis size or morphological development of the testis, suggesting that gonadotrophins regulate testicular development during a 'critical window' late in gestation. Taken together, these studies provide convincing evidence that GnRH plays a central role in the ontogeny and regulation of pituitary-gonadal function during fetal life.

Effects of LH-RH infusion, castration and cryptorchidism on gonadotrophin and testosterone secretion in developing rams

Summary. The relationship between the pituitary gland and testis in rams was studied from birth to sexual maturity. The concentrations of LH, FSH and testosterone increased between 5 and 7 weeks of age; the rise was not correlated with any specific cytological change in the testis. An augmented pituitary response to LH-RH was demonstrated as levels of gonadotrophin increased. It is unclear whether this change in sensitivity plays a role in initiation of the pubertal process because Sertoli cell maturation, the earliest detectable change in the seminiferous epithelium, occurs between 17 and 21 weeks of age. Spermatocytes were first seen in biopsies taken at 31–36 weeks and spermatogenesis was established fully by 45 weeks. This second phase of testicular development was characterized by increases in prolactin, testosterone and LH. Leydig cells previously difficult to identify became recognizable at the time of sexual maturation.

Aging and the Male Reproductive System.

This narrative review presents an overview of current knowledge on fertility and reproductive hormone changes in aging men, the factors driving and modulating these changes, their clinical consequences, and the benefits and risks of testosterone (T) therapy. Aging is accompanied by moderate decline of gamete quality and fertility. Population mean levels show a mild total T decline, an SHBG increase, a steeper free T decline, and a moderate LH increase with important contribution of comorbidities (e.g., obesity) to these changes. Sexual symptoms and lower hematocrit are associated with low T and are partly responsive to T therapy. The relationship of serum T with body composition and metabolic health is bidirectional; limited beneficial effects of T therapy on body composition have only marginal effects on metabolic health and physical function. Skeletal changes are associated primarily with estradiol and SHBG. Cognitive decline is not consistently linked to low T and is not improved by T therapy. Although limited evidence links moderate androgen decline with depressive symptoms, T therapy has small beneficial effects on mood, depressive symptoms, and vitality in elderly patients with low T. Suboptimal T (and/or DHT) has been associated with increased risk of stroke, but not of ischemic heart disease, whereas an association with mortality probably reflects that low T is a marker of poor health. Globally, neither severity of clinical consequences attributable to low T nor the nature and magnitude of beneficial treatment effects justify the concept of some broadly applied "T replacement therapy" in older men with low T. Moreover, long-term safety of T therapy is not established.

The Contribution of the Circadian Gene Bmal1 to Female Fertility and the Generation of the Preovulatory Luteinizing Hormone Surge.

In rodents, the preovulatory LH surge is temporally gated, but the timing cue is unknown. Estrogen primes neurons in the anteroventral periventricular nucleus (AVPV) to secrete kisspeptin, which potently activates GnRH neurons to release GnRH, eliciting a surge of LH to induce ovulation. Deletion of the circadian clock gene Bmal1 results in infertility. Previous studies have found that Bmal1 knockout (KO) females do not display an LH surge at any time of day. We sought to determine whether neuroendocrine disruption contributes to the absence of the LH surge. Because Kiss1 expression in the AVPV is critical for regulating ovulation, we hypothesized that this population is disrupted in Bmal1 KO females. However, we found an appropriate rise in AVPV Kiss1 and Fos mRNA at the time of lights out in ovariectomized estrogen-treated animals, despite the absence of a measureable increase in LH. Furthermore, Bmal1 KO females have significantly increased LH response to kiss-10 administration, although the LH response to GnRH was unchanged. We then created Kiss1- and GnRH-specific Bmal1 KO mice to examine whether Bmal1 expression is necessary within either kisspeptin or GnRH neurons. We detected no significant differences in any measured reproductive parameter. Our results indicate that disruption of the hypothalamic regulation of fertility in the Bmal1 KO females is not dependent on endogenous clocks within either the GnRH or kisspeptin neurons.

Features of the metabolic syndrome in late adolescence are associated with impaired testicular function at 20 years of age.

Features of the metabolic syndrome in late adolescence are associated with impaired testicular function at 20 years of age.

Effect of Phitofert® on testicular function of vasectomized mature mice

This study was designed to evaluate the role of Phitofert® in the improvement of testes function and attenuating DNA fragmentation in vasectomized and healthy adult mice. Twenty four adult male breed such Albino mice were randomly and equally divided into four groups (G1, G2, G3 and G4). Mice were treated for 35days as follows: the G1 mice were non-vasectomized and given DW and served as controls, mice in G2 were non-vasectomized and given Phitofert® daily with a dose of 0.035mg/kg BW, mice in G3 were vasectomized without treatment while the mice in G4 were vasectomized and given same dose of Phitofert®. At the end of the experiment, fasting blood samples were collected by cardiac puncture for measuring LH and FSH hormones concentrations and section from testes were taken tomeasure the number of Leydig cells and diameter of semniferous tubules. The results of current data showed significant increase of serum LH and FSH concentrations in G2 healthy treated group as compared with control and other groups. Also, the treatment of G4 vasectomized mice with Phitofert® caused significant increase in serum concentration of the above hormones as compared with G3 vasectomized non-treated. The study showed that DNA fragmentation that resulted from the G2 healthy treated mice were lower than that obtained from the control group and other vasectomized mice (G3, G4). The diameters of the seminiferous tubules and the numbers of Leydig cells showed significant increase in healthy and vasectomized treated group (G1, G4) as compared with G3 vasectomized non-treated group. It was concluded that healthy and vasectomized treated of adult male mice with Phitofert® lead to clear improvement of level of reproductive hormones.

Role of irisin administration in modulating testicular function in adult obese albino rats

AbstractBackground: Irisin, exercise-induced myokine, is known to improve obesity-induced metabolic disturbances by enhanc-ing thermogenesis and increasing energy expenditure. Recent studies have reported that irisin promoted cell proliferation and protected cells from apoptosis. However, the effects of irisin on testicular function in obese rats is still to be investi-gated.Aim of the Study: This study was designed to explore the probable effects of irisin treatment on modulating the adverse effects of obesity on testicular functions with a trial to clarify some of the possible underlying mechanisms.Material and Methods: Thirty adult male wistar albino rats (200±30g) were divided equally into 3 groups, group (I): Saline-vehicle treated normal fed group, group (II): Saline-vehicle treated rats were fed a High Fat Diet (HFD) for 10 weeks. Group (III): Irisin treated HFD-fed rats for 10 weeks and irisin was injected subcutaneously (150ml daily for 4 weeks). Rats were weighed and serum levels of glucose, insulin, lipid profile, FSH, LH, estradiol & testosterone levels, epididymal sperm count and motility, testicular Malondiald-hyde (MDA) level, Superoxide Dismutase (SOD), Catalase (CAT), Glutathione Peroxidase (GPX) activities, testicular and epidydimal fat weights were estimated. Testicular his-topathological study was done as well.Results: There was a significant increase in the Body Mass Index (BMI) together with deterioration of metabolic, gonadal functions and testicular oxidant/antio-oxidant system in group (II), while exogenous irisin administration in group (III) resulted in: Significant decrease in BMI, insulin resistance with improvement in lipid profile, significant increase in serum FSH, LH & testosterone, sperm count and motility in male rats. Together with marked improvement in the testicular histoarchitecture. Additionally, there was a significant increase in testicular SOD, CAT & GPX activities. However, it signif-icantly decreased testicular MDA level.Conclusions: It could be suggested that irisin has a po-tential positive role against obesity-induced testicular dys-function, which may be due to its role in maintenance of glucose & insulin homeostasis, its antioxidant properties and/ or maintenance of gonadal hormonal function via indirect and/or direct effects on the gonads.

Role of Irisin Administration in Modulating Testicular Function in Adult Obese Albino Rats

AbstractBackground: Irisin, exercise-induced myokine, is known to improve obesity-induced metabolic disturbances by enhanc-ing thermogenesis and increasing energy expenditure. Recent studies have reported that irisin promoted cell proliferation and protected cells from apoptosis. However, the effects of irisin on testicular function in obese rats is still to be investi-gated.Aim of the Study: This study was designed to explore the probable effects of irisin treatment on modulating the adverse effects of obesity on testicular functions with a trial to clarify some of the possible underlying mechanisms.Material and Methods: Thirty adult male wistar albino rats (200±30g) were divided equally into 3 groups, group (I): Saline-vehicle treated normal fed group, group (II): Saline-vehicle treated rats were fed a High Fat Diet (HFD) for 10 weeks. Group (III): Irisin treated HFD-fed rats for 10 weeks and irisin was injected subcutaneously (150ml daily for 4 weeks). Rats were weighed and serum levels of glucose, insulin, lipid profile, FSH, LH, estradiol & testosterone levels, epididymal sperm count and motility, testicular Malondiald-hyde (MDA) level, Superoxide Dismutase (SOD), Catalase (CAT), Glutathione Peroxidase (GPX) activities, testicular and epidydimal fat weights were estimated. Testicular his-topathological study was done as well.Results: There was a significant increase in the Body Mass Index (BMI) together with deterioration of metabolic, gonadal functions and testicular oxidant/antio-oxidant system in group (II), while exogenous irisin administration in group (III) resulted in: Significant decrease in BMI, insulin resistance with improvement in lipid profile, significant increase in serum FSH, LH & testosterone, sperm count and motility in male rats. Together with marked improvement in the testicular histoarchitecture. Additionally, there was a significant increase in testicular SOD, CAT & GPX activities. However, it signif-icantly decreased testicular MDA level.Conclusions: It could be suggested that irisin has a po-tential positive role against obesity-induced testicular dys-function, which may be due to its role in maintenance of glucose & insulin homeostasis, its antioxidant properties and/ or maintenance of gonadal hormonal function via indirect and/or direct effects on the gonads.

Effects of icariin on ovarian function in d-galactose-induced aging mice

In this study, effects of icariin (Ica) on were examined in a mouse model of d-galactose (D-gal)-induced ovarian aging. Kunming white mice were divided into three groups: aging group induced with D-gal, experiment group treated with Ica at low (50 mg/kg), middle (100 mg/kg) and high (200 mg/kg) concentrations, and control group with no treatment. Ovarian histomorphology, serum FSH, LH and E2 levels, and reproductive function were compared among the groups. Ovarian expression of Amh, Bax and Bcl-2 was examined by qPCR and western blotting. Our results showed that diameters of secondary and tertiary follicles were significantly reduced in the aging group when compared with control group (P < 0.01), and were restored to normal in Ica 100 and Ica 200 treatment groups. The diameter of atretic follicles was significantly smaller in the aging group compared with control group and Ica 200 treatment group (P < 0.05). The proportion of secondary and atretic follicles was higher in the aging group compared with control group, Ica 100 and 200 treatment groups, whereas the proportion of tertiary and mature follicles was reduced in the aging group versus control, Ica 100 and 200 groups. The aging group lacked mature follicles, whereas Ica treatment induced mature follicle development. Primary and secondary follicles exhibited similar theca cell numbers and theca interna and externa cell layers in all groups examined, whereas theca interna and externa cell layers were decreased and increased, respectively, in tertiary follicles of aging group compared with control and I 200 groups. In the aging group, FSH and LH levels were significantly higher than those in control and Ica 200 groups (P < 0.05), and the E2 level was significantly reduced compared with control (P < 0.01), Ica 200 (P < 0.01), and Ica 100 (P < 0.05) groups. Serum hormone levels were equivalent in the control, Ica 100 and Ica 200 groups. The pregnancy rate was reduced in the aging group compared with other groups. The average litter size per birth, birth litter weight, and weaning weight of litters were all significantly lower in the aging group compared with control, Ica 100 and 200 groups (P < 0.05). The ovarian expression of AMH and Bcl-2 mRNA was significantly reduced in the aging group compared with those in control and Ica-treated groups (P < 0.01). In contrast, Bax expression was significantly higher in the aging group compared with all other groups (P < 0.01), and the Bcl-2/Bax ratio was markedly reduced in aging group compared with control, Ica 100 and 200 groups (P < 0.01), and Ica 50 group (P < 0.05). Ovarian expression of AMH protein was elevated in the Ica 100 group compared with the aging, control and Ica 50 groups (P < 0.01) and Ica 200 group (P < 0.05). Ovarian Bcl-2 protein levels and the Bcl-2/Bax ratio were significantly higher in the Ica 100 group than those in the Ica 50, 200 and aging groups (P < 0.05), and were similar or reduced (P < 0.05), respectively, compared to those in control group. Ovarian Bax expression was similar in each group. These findings suggest that Ica can improve ovarian follicular development, inhibit follicular atresia, decrease FSH and LH levels and increase E2, upregulate ovarian AMH expression and increase the Bcl-2/Bax ratio in aging mice. Therefore, Ica can partially restore ovarian function of aging mice and enhance their fertility. Optimal reproductive effects were obtained with the Ica 100 group.

Impact of Bariatric Surgery on Male Sex Hormones and Sperm Quality: a Systematic Review and Meta-Analysis.

This systematic review and meta-analysis aims to establish the effects of bariatric surgery on male sex hormones, sperm parameters, and sexual function. We searched MEDLINE, EMBASE, Web of Science, and Scopus from database inception through June 2018. Articles were eligible for inclusion if they examined the effect of bariatric surgery on male sex hormones and sperm parameters in patients with obesity. Primary outcomes of interest were sex hormones and sperm quality. Secondary outcome was sexual function (International Index of Erectile Function (IIEF) score). Pooled estimates were calculated using random effects meta-analysis. A total of 28 cohort studies with 1022 patients were identified from 3896 potentially relevant citations. Both free and calculated testosterone levels were significantly increased after bariatric surgery (mean difference (MD) - 7.47nM, 95% CI - 8.62 to - 6.31, p < 0.001 and MD - 0.05nM, 95% CI - 0.07 to - 0.02, p < 0.001, respectively). Consistent with the increase in testosterone, LH, FSH, and SHBG levels were also significantly increased after surgery. In contrast, free and total estradiol and prolactin levels were significantly decreased after bariatric surgery. From studies that reported the IIEF score, bariatric surgery led to a significant increase in erectile function after surgery (MD - 0.46, 95% CI - 0.89 to - 0.02, p = 0.04). However, bariatric surgery did not affect sperm quality, DHEA, androstenedione, and inhibin B levels. Sustained weight-loss induced by bariatric surgery had a significant effect on increasing male sex hormones and decreasing female sex hormones in male patients with obesity. However, sperm quality and function were not improved after surgery.

Decrease in semen quality and Leydig cell function in infertile men: a longitudinal study.

Are infertile men with reduced semen quality at risk of a further decrease in testicular function? Infertile men with severely reduced semen quality risk further deterioration of semen quality 15 years after treatment for infertility, and a lower baseline sperm concentration was associated with a more pronounced increase in LH and decrease in testosterone/LH ratio at follow-up. Male factors account for up to 50% of human infertility. The most common finding is spermatogenic failure (SgF) yet the life course of semen quality and testosterone production in such men has not been described. A follow-up study of men with SgF was performed 15 years after the initial infertility assessment between January 1995 and December 2000. Hospital records were used to identify potential participants in the study. A total of 137 men with primary male infertility due to SgF and 70 controls with good semen quality from couples with female factor infertility who attended a tertiary referral centre were included: the participation rate was 31% and 26%, respectively. The men provided semen samples and underwent a physical examination. Blood samples were taken to measure levels of reproductive hormones (FSH, LH, testosterone, sex hormone-binding globulin, estradiol and inhibin B). Current results were compared with results from the initial assessments. At the time of follow up the SgF men had significantly lower Leydig cell capacity than the control group as well as much lower semen quality. For the SgF men, between baseline sampling and follow up, the median sperm concentration decreased from 1.9 to 0.6 mill/ml and total sperm count from 7.7 to 2.0 million (P = 0.019 and 0.012, respectively), and 10% developed azoospermia. Calculated free testosterone (cFT), but not total testosterone (tT) decreased in the SgF group by ~0.6% (95% CI 0.1-1.2%) per year. In the SgF group, LH increased by 1.6% (CI 0.9-2.3%) annually, and consequently tT/LH and cFT/LH ratios had decreased by 1.3% (CI 0.5-2.1) and 2.1% (CI 1.2-3.0%), respectively. The increase in LH and the decreases in tT/LH and cFT/LH ratios were more pronounced in men with lower baseline sperm concentrations. We consider the case group as representative of infertile men not in need of testosterone treatment at baseline investigation, but do not have information on those that chose not to participate in the follow-up study. There were alterations in some hormone analysis methods during the follow-up period that may introduce uncertainty in interpretation of long-term changes in hormone levels despite rigorous quality control. The validity of the control group suffers from a lack of hormone values at baseline. Also, at follow-up, for practical reasons only one semen sample could be obtained, which makes the effect estimate more uncertain and there is a risk of non-differential misclassification. Without being able to predict individual outcomes, it is prudent to consider sperm cryopreservation or advise not to postpone fertility treatment when men present with infertility due to impaired semen quality. Whether partly compensated Leydig cell insufficiency in men with SgF will eventually develop into overt testosterone deficiency cannot be determined from our study. Aase and Einar Danielsen (Grant no. 10-001053), Nordic Research Committee (Grant no. 5109), The Kirsten and Freddie Johansen Fund, and Rigshospitalet's Research Fund (grant no. R24-A812). There are no competing interests.

Variations in the levels of LH and FSH hormonal status in Obesity condition

The study aims to access the variations in the levels of LH and FSH hormonal status in Obesity condition. Nowadays obesity is a very common condition we can see in most of the people living in developing countries. It causes so many other health problems along with their regular lifestyle habituates. In that infertility is one of the important problems occurs due to obesity by altering the hormonal levels in the female population. 30 obese patients and 30 healthy individuals from the OP of Saveetha Dental College. Serum samples were analysed for their hormonal status by using a kit method in auto analyser. There is a significant increase in LH (12.88 ± 6.69) and FSH (9.55 ± 4.8) as well as their ratio (LH/FSH- 1.93 ± 2.13) by the influence of obesity on them. The study states that a high risk of obesity may lead to hormonal imbalance which may lead to infertility in obese female patients.

Kisspeptin Modulates Luteinizing Hormone Release and Ovarian Follicular Dynamics in Pre-pubertal and Adult Murrah Buffaloes.

Kisspeptin is a neuropeptide that governs the reproductive axis upstream to GnRH. We wanted to study whether kisspeptin modulates plasma LH and FSH levels and ovarian follicular dynamics in buffaloes and whether kisspeptin can be used for fixed time artificial insemination (FTAI). We carried out these studies in comparison with buserelin, a potent GnRH agonist. Kisspeptin dose-dependently increased plasma LH levels. However, the kisspeptin-induced increase in LH was short-lived as the peak reached in 15–30 min returned to basal values by 1–2 h. The kisspeptin-induced increase in LH level was less compared to buserelin-induced increase in LH level which sustained over time. Kisspeptin did not enhance FSH release while buserelin resulted in a gradual increase over time. LH response to repeated injections of kisspeptin was greater than that induced by buserelin. While buserelin induced an increase in the number of follicles, kisspeptin induced an increase in the growth rate of the follicle. In adult cycling animals, while both the drugs increased plasma LH levels, the increase was greater in buserelin group compared to kisspeptin group. In contrast to the findings in pre-pubertal animals, kisspeptin induced an increase in both the number as well as the size of follicles compared to buserelin. Our studies on oestrus synchronization, using either kisspeptin-PGF2α-kisspeptin protocol or buserelin-PGF2α-buserelin Ovsynch protocol on day 0, 7, and 9, respectively, revealed that kisspeptin increased the number of follicles at wave emergence and the diameter of dominant follicle after 2nd dose of drug, the oestrus response rate and duration of oestrus, compared to buserelin. However, conception rate was not significantly different among the groups. From our studies, it appears that Kp and Buserelin differentially modulate follicular dynamics depending on the reproductive age of the animals.However, studies in a larger herd are required to confirm whether kisspeptin can be used for oestrous synchronization in buffaloes.

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

Anti-Müllerian hormone (AMH) has gained increasing interest as a biomarker for assessment of gonadal activity. The ability to predict the ovarian follicular reserve of prepubertal female horses (fillies) or to identify stallions with testicular pathologies already during their prepubertal life has not been analyzed so far. Both would help to select fertile horses and reduce costs associated with keeping animals. The objectives of the present study were to (1) assess AMH, LH, FSH, progesterone (females) and testosterone (males) dynamics in prepubertal horses from birth onwards and (2) determine whether AMH concentrations detected in plasma of prepubertal female and male horses are correlated with postpubertal gonadal development. Warmblood foals (n = 30, 14 females, 10 normal males and 6 males with abnormal testicular development) born between February and May of two consecutive years (n = 28 in the first year and n = 2 the next year), were included in the study. Information on gestational length, parity of the dam and placental weight was collected for all foals. Blood samples for hormone analysis were collected from birth onwards every four weeks up to the age of one year. At two years, blood samples were collected on the day when antral follicle count (AFC) and total testicular volume (TTV) were assessed. AMH was detectable in the plasma of all animals from birth onwards and its concentration was significantly higher (P < .001) in males than in females, regardless of testicular development. In males, AMH and testosterone concentration were similar for all animals during the first year of life, regardless of testicular development. At two years, AMH concentration was higher (P < .05) in males with abnormal testicular development than in those with normal testes. In females, AMH concentration at two years was correlated with AMH concentration at birth (P < .05) and with AFC (P < .001). At birth, LH concentration was lower (P < .05) in stallions with abnormal testes (0.3 ± 0.2 ng/ml) than in controls (0.6 ± 0.2 ng/ml). A high negative correlation between AMH concentration and gestation length was observed in males during the first eight weeks of life (P < .01, r = −0.64 to −0.71). Elevated progesterone concentrations over 1 ng/ml were observed in several females starting with 20 weeks of age. This was paralleled by an increase in AMH concentration and was preceded by FSH and LH increases. In conclusion, AMH determination can be reliably used from two years onwards to identify stallions with abnormal testicular development, but it is inconclusive before puberty. In female horses, determination of AMH concentration at a prepubertal age allows for prediction of AMH and AFC after puberty. We suggest that premature luteinisation occurs before the onset of puberty in female horses and that LH secretion in the perinatal period is involved in testicular development and descent in the horse.