Luteinizing Hormone Receptor Research Articles

Growth hormone in fertility and infertility: Mechanisms of action and clinical applications

Secreted by the anterior pituitary gland, growth hormone (GH) is a peptide that plays a critical role in regulating cell growth, development, and metabolism in multiple targeted tissues. Studies have shown that GH and its functional receptor are also expressed in the female reproductive system, including the ovaries and uterus. The experimental data suggest putative roles for GH and insulin-like growth factor 1 (IGF-1, induced by GH activity) signaling in the direct control of multiple reproductive functions, including activation of primordial follicles, folliculogenesis, ovarian steroidogenesis, oocyte maturation, and embryo implantation. In addition, GH enhances granulosa cell responsiveness to gonadotropin by upregulating the expression of gonadotropin receptors (follicle-stimulating hormone receptor and luteinizing hormone receptor), indicating crosstalk between this ovarian regulator and the endocrine signaling system. Notably, natural gene mutation of GH and the age-related decline in GH levels may have a detrimental effect on female reproductive function, leading to several reproductive pathologies, such as diminished ovarian reserve, poor ovarian response during assisted reproductive technology (ART), and implantation failure. Association studies using clinical samples showed that mature GH peptide is present in human follicular fluid, and the concentration of GH in this fluid is positively correlated with oocyte quality and the subsequent embryo morphology and cleavage rate. Furthermore, the results obtained from animal experiments and human samples indicate that supplementation with GH in the in vitro culture system increases steroid hormone production, prevents cell apoptosis, and enhances oocyte maturation and embryo quality. The uterine endometrium is another GH target site, as GH promotes endometrial receptivity and pregnancy by facilitating the implantation process, and the targeted depletion of GH receptors in mice results in fewer uterine implantation sites. Although still controversial, the administration of GH during ovarian stimulation alleviates age-related decreases in ART efficiency, including the number of oocytes retrieved, fertilization rate, embryo quality, implantation rate, pregnancy rate, and live birth rate, especially in patients with poor ovarian response and recurrent implantation failure.

Understanding the effects of sustained supraphysiologic concentrations of luteinizing hormone in gonadectomized dogs: What we know and what we still need to learn

Removal of the gonads with surgical sterilization results in a loss of negative feedback to the hypothalamus and anterior pituitary. The sustained supraphysiologic luteinizing hormone (LH) concentrations in gonadectomized dogs can significantly alter organ function and even induce neoplastic changes. For example, gonad removal has a profound effect on thyroid function and is reported to be the most significant cause for the development of hypothyroidism in dogs. Thirty percent more gonadectomized dogs develop hypothyroidism compared with intact dogs. Within the canine thyroid, LH receptors are co-localized with thyroid stimulating hormone (TSH) receptors. Continuous LH receptor activation in gonadectomized dogs may interfere TSH receptor function by consuming second messengers involved in G-protein receptor cell signaling, preventing the action of TSH when it binds to its receptor in the thyroid, resulting in hypothyroidism. The incidence of anterior cruciate ligament ruptures is significantly increased following gonad removal independent of breed, sex, weight or body condition. Luteinizing hormone receptors are expressed in the cruciate ligament and continuous LH receptor activation may increase laxity in these ligaments, resulting in joint instability. Both male and female gonadectomized dogs are at a significantly increased risk for lymphoma and hemangiosarcoma. Luteinizing hormone receptors are also abundant in these tissues. Research in four canine hemangiosarcoma cell lines found that LH receptor activation induces cell proliferation. In addition, research in three canine T-cell lymphoma cell lines found that LH receptor activation induces cell proliferation, adhesion, and invasion as well as increases LH receptor expression. Research is needed to determine if LH reducing strategies using gonadotropin releasing hormone agonists will increase remission times in gonadectomized dogs with LH receptor-positive tumors. In conclusion, among the non-reproductive functions of gonads, suppression of LH secretion and resulting LH receptor overexpression appear necessary in maintaining endocrine, musculoskeletal, and anti-neoplastic health.

RF33 | PSUN04 Luteinizing Hormone-Chorionic Gonadotrophin Receptor in Pheochromocytomas

Abstract Background Pheochromocytomas and paragangliomas (PPGL) are catecholamine-producing neuroendocrine tumors that arise from the adrenal medulla or ganglia and display the highest heritability rate among all human tumours. Genomic analyses allowed identification of molecular subgroups of PPGL which are organized into 2 main clusters. Cluster 1 contains SDHx- and VHL-mutated tumors which do not produce epinephrine while cluster 2 includes the epinephrine-secreting PPGL related to RET, NF1, TMEM127 and MAX mutations. PPGL must be early diagnosed and treated to prevent adrenergic crises which can be life-threatening. Detection of PPGL is particularly important during pregnancy since PPGL are associated with a high risk of either maternal or fetal complications in this context. Reciprocally, pregnancy can favor adrenergic crises in patients with previously undiagnosed or silent pheochromocytoma (PCC). These dangerous events can be triggered by tumor compression resulting from fetus growth, or labor and delivery. However, it is known that surges in plasma catecholamines may also occur during early gestation suggesting that pregnancy may also activate the secretory activity of PPGL through the involvement of non-mechanical factors, such as gestational hormones. Methods In the present study, we report a case of silent PCC revealed in a pregnant woman by life-threatening adrenergic myocarditis occurring at 31 weeks of gestation. Analysis of PPGL susceptibility genes showed the presence of a heterozygous germline RET variant of uncertain significance. The fact that the first symptoms of catecholamine excess had appeared during the first trimester of pregnancy led us to conduct in vitro studies to investigate the effects of estradiol and the pregnancy hormone chorionic gonadotropin (hCG) on epinephrine secretion by cultured cells derived from the resected patient's tumor. Expression of luteinizing hormone/chorionic gonadotropin receptor (LHCGR) was searched for in the tumor and an additional series of 12 PCC by RT-qPCR and immunohistochemistry. LHCGR expression was also analyzed in silico in the PPGL cohorts of the COMETE and The Cancer Genome Atlas (TCGA) databases. Results hCG stimulated epinephrine secretion by primary cultured PCC cells. The tumor expressed the LHCG receptor, which was colocalized with catecholamine-producing enzymes. A similar expression pattern of the LHCG receptor was also observed in 5 out of a series of 12 PCCs. Moreover, in silico studies revealed that PPGL display the highest expression levels of LHCGR mRNA among the 32 solid tumor types of TCGA cohort. Interestingly, expression of LHCGR was higher in cluster 2 than in cluster 1 PPGL. Conclusion We show that PCC can express functional LHCG receptor. Consequently, pregnancy may activate catecholamine production by previously silent pheochromocytoma as early as the first trimester of gestation especially in women with gene mutations that predispose to cluster 2 epinephrine-secreting PCC. Presentation: Sunday, June 12, 2022 12:30 p.m. - 2:30 p.m., Monday, June 13, 2022 1:18 p.m. - 1:23 p.m.

Analysis of LH Receptor Expression in the Testes of Infertile Azoospermic Non-Obstructive (NOA) Men at High Serum Prolactin Concentrations

Analysis of LH Receptor Expression in the Testes of Infertile Azoospermic Non-Obstructive (NOA) Men at High Serum Prolactin Concentrations

Effects of Different Oxytocin and Temperature on Reproductive Activity in Nile tilapia (Oreochromis niloticus): Based on Sex Steroid Hormone and GtHR Gene Expression

Luteinizing hormone receptor (LHR) and follicle-stimulating hormone receptor (FSHR) belong to the gonadotropic hormone receptors (GtHR), which are highly expressed in fish gonads and participate in the regulation of fish reproductive activities. Fish gonadal development and gamete maturation are not only regulated by their BPG axis but also affected by natural environmental factors (such as temperature, salinity, pH, nutrients, light, etc.). Nile tilapia (Oreochromis niloticus) is a farmed fish with a short reproductive cycle, fast growth, and high economic value. To study the relationship between gonadotropic hormone receptors (GtHR) and the reproductive activity of Nile tilapia, different oxytocin injection experiments and different temperature treatment experiments were set up, and the expression changes of the GtHR gene in the gonads and the concentration changes of the estradiol (E2) in the female serum and testosterone (T) in the male serum were determined employing a quantitative RT-PCR assay and enzyme-linked immunosorbent assay (ELISA), respectively. After the injection of oxytocin, with the change of E2 in females and T in males, the FSHR showed an expression pattern of first increase, then decrease, and the LHR showed an expression pattern of first increase, then decrease, and finally increase in the gonads, and the expression level of FSHR and LHR in the injection group was significantly higher than that in the control group at multiple time points; in addition, the expression level of FSHR and LHR in the oxytocin-combination injection group was higher than that in the single injection group. During 28 days of treatment at different temperatures, the sex steroid hormones and GtHR genes also showed regular changes, and the relationship between each group was 28 °C > 32 °C > 24 °C at most time points. According to the research results, it is speculated that FSHR and LHR play an important role in the development of Nile tilapia gonads and participate in the reproductive activities of Nile tilapia. By comparing and analyzing the changes in the sex steroid hormones and GtHR genes in each experimental group, it is speculated that different oxytocin injections could affect the expression of FSHR and LHR genes in Nile tilapia, and the combined effect of oxytocin was better than single oxytocin; the optimum temperature for the reproduction of Nile tilapia is between 28–32 °C. This study provides a theoretical basis for further elucidating the physiological functions and molecular mechanisms of FSHR and LHR and also provides a reference for the research of reproductive regulation in Nile tilapia.

Luteinizing Hormone Receptor Expression in Neoplastic Mast Cells Is Increased in Spayed and Neutered Dogs.

Luteinizing hormone receptors (LHRs) are expressed in canine lymphoma and hemangiosarcoma. We hypothesized that LHR would be expressed in canine mast cell tumors (MCTs) and that more neoplastic mast cells would express LHR in gonadectomized dogs compared with intact dogs. Eleven archived formalin-fixed paraffin-embedded cutaneous MCT tissue sections were processed using routine immunohistochemistry. For both the KIT protein and LHR, the percentage of positive cells for each staining pattern (I-III) was calculated. A Student's t test was used to compare the total percentage of positive cells expressing LHR and KIT in intact and gonadectomized dogs. A one-way analysis of variance was used to compare the percentage of cells within each staining pattern for LHR and KIT in intact and gonadectomized dogs. All MCT expressed LHR. MCT from gonadectomized dogs had a significantly higher percentage of LHR-positive mast cells (84.2 ± 8.7%) compared with MCTs from intact dogs (64.3 ± 4.2%). This is the first study to demonstrate the expression of LHR in canine MCTs and to report that LHR expression is increased in neoplastic mast cells from gonadectomized dogs compared with intact dogs. Future studies are planned to evaluate the functionality of the LHR in canine neoplastic mast cells.

A Possible Mechanism for Double-Yolked Eggs in the Early Stage of Egg-Laying in Zhedong White Goose-Function of IGF1 and LHR Signaling.

The cause of double-yolk (DY) egg production in birds is unclear, but it is related to body weight and adiposity. We explored the causes of the high proportion (up to 26%) of DY eggs in the first clutch of Zhedong white geese. We recorded the egg production of Zhedong white geese during the first egg-laying cycle and counted the proportion of DY eggs. We found that 30% of geese had 3 sets of double or triple follicles of the same diameter in the abdomen, which was close to the DY egg rate. In addition, the mRNA expression levels of the steroidogenic acute regulatory protein (StAR) and luteinizing hormone receptor (LHR) genes in granulosa cells were similar within the same set of follicles. Furthermore, the IGF1 concentration in geese that had at least 3 sets of follicles of the same diameter was significantly higher than that in birds with 0-1 set of follicles of the same diameter. Thus, we proposed that, in the first egg-laying stage of geese, high plasma concentrations of IGF1 stimulate the development of pre-hierarchal follicles and cause more than one follicle to be selected at the same time, mature at the same rate under the same gonadotrophin milieu, and ovulate at the same time to produce DY eggs.

Divergent G-protein signal control at the very early endosome (VEE) from the dually coupled luteinizing hormone receptor (LHR)

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

Cytogenetic and molecular insight into the genetic background of disorders of sex development in seventeen cats

The genetic background of feline disorders of sex development (DSDs) is poorly understood. We performed comprehensive cytogenetic, molecular, and histological studies of 17 cats with abnormal external genitalia, unusual behavior, or tricolor coats (atypical in males). The DSD phenotype of three cats was associated with sex chromosome abnormalities: X/Y translocation (38,XXSRY+), 37,X/38,XY mosaicism, and XX/XY leukocyte chimerism. The remaining 14 affected cats were classified as XY DSD (SRY-positive). In this group and 38 normal males, we analyzed a priori selected candidate genes (SRY, TAC3, CYP11B1 and LHCGR). Only a previously reported nonpathogenic variant was found in SRY. Moreover, SRY gene copy number was determined, and three variants were observed: 6, 5 (modal), and 4 copies in a single DSD case. The known variants in TAC3 and CYP11B1, responsible for testicular hypoplasia, persistent primary dentition or congenital adrenal hyperplasia, were not found in the study group. Nine novel polymorphisms were identified in the LHCGR gene, one of which, a potentially regulatory indel variant in 5′UTR, was significantly associated (p = 0.0467) with XY DSD. Our report confirmed that abnormalities of sex chromosomes are important causes of feline DSDs. We also showed that the indel variant of LHCGR can be considered a promising marker associated with XY DSD phenotype.

Umbilical Cord-Derived Mesenchymal Stem Cells for the Treatment of Infertility Due to Premature Ovarian Failure.

Females belonging to the reproductive age group may face challenges regarding infertility or miscarriage due to conditions such as premature ovarian failure (POF). It is the conditionthat happens when a female's ovaries stop working before she is 40. The majority of the causes of POF cases are idiopathic. Other reasons includegenetic disorders (Turner's syndrome, bone morphogenetic protein 15 (BMP15) mutation, galactosemia, mutation of forkhead box protein L2 (FOXL2), growth differentiation factor-9 (GDF9), mutation of luteinizing hormone (LH) and follicle-stimulating hormonereceptors (FSHR), etc.), enzymatic mutation such as aromatase, autoimmune disorders (Addison's disease, vitiligo, systemic lupus erythematosus, myasthenia gravis, autoimmune thyroiditis, autoimmune polyglandular syndrome, etc.), vaccination, and environmental factors (cigarette smoking, toxins, and infections). Manyattempts have been made to treat POF by various methods. Some of the methods of treatment include hormone replacement therapy (HRT), melatonin therapy, dehydroepiandrosterone (DHEA) therapy, and stem cell therapy. Stem cell therapy has proven to be the most efficient form for treating POF as compared to all other options. Umbilical cord-derived mesenchymal stem cells (UC-MSCs) are the best among the other sources of mesenchymal stem cells (MSCs) for the treatment of POF as they have a painless extraction procedure. They have a tremendous capacity for self-repair and regeneration, which helps them in restoring degenerated ovaries. This review includes informationon the causes of POF, its efficacious therapeutic approaches, and the impact of transplantation of human umbilical cord mesenchymal stem cells (hUCMSCs) as an option for the therapy of POF. Numerous studies conducted on stem cell therapy provethat it is an effective approach for the treatment of sterility.

The Corticosterone-Glucocorticoid Receptor-AP1/CREB Axis Inhibits the Luteinizing Hormone Receptor Expression in Mouse Granulosa Cells.

Under stress conditions, luteinizing hormone (LH)-mediated ovulation is inhibited, resulting in insufficient oocyte production and excretion during follicular development. When the body is stressed, a large amount of corticosterone (CORT) is generated, which will lead to a disorder of the body’s endocrine system and damage to the body. Our previous work showed that CORT can block follicular development in mice. Since LH acts through binding with the luteinizing hormone receptor (Lhcgr), the present study aimed to investigate whether and how corticosterone (CORT) influences Lhcgr expression in mouse ovarian granulosa cells (GCs). For this purpose, three-week-old ICR female mice were injected intraperitoneally with pregnant mare serum gonadotropin (PMSG). In addition, the treatment group was injected with CORT (1 mg/mouse) at intervals of 8 h and the control group was injected with the same volume of methyl sulfoxide (DMSO). GCs were collected at 24 h, 48 h, and 55 h after PMSG injection. For in vitro experiments, the mouse GCs obtained from healthy follicles were treated with CORT alone, or together with inhibitors against the glucocorticoid receptor (Nr3c1). The results showed that the CORT caused a downregulation of Lhcgr expression in GCs, which was accompanied by impaired cell viability. Moreover, the effect of the CORT was mediated by binding to its receptor (Nr3c1) in GCs. Further investigation revealed that Nr3c1 might regulate the transcription of Lhcgr through inhibiting the expression of Lhcgr transcription factors, including AP1 and Creb. Taken together, our findings suggested a possible mechanism of CORT-induced anovulation involving the inhibition of Lhcgr expression in GCs by the CORT–Nr3c1–AP1/Creb axis.

Application of an Allosteric Agonist of the Luteinizing Hormone Receptor for Reducing the Effective Dose of Gonadotropin in the Treatment of Androgen Deficiency in Rats with Type 1 Diabetes

In type 1 diabetes mellitus, the impaired testosterone synthesis in the testes leads to androgen deficiency. The long-term application of high gonadotropin doses for its correction decreases the sensitivity of luteinizing hormone/human chorionic gonadotropin (LH/hCG) receptors in Leydig cells to the endogenous gonadotropins. The aim of this work was to study the effect of a 3-day treatment of male Wistar rats with streptozotocin type 1 diabetes with the 5-amino-N-tert-butyl-2-(methylsulfanyl)-4-(3-(nicotinamido) phenyl)thieno[2,3-d]pyrimidine-6-carboxamide allosteric LH/hCG receptor agonist (TP03, 15 mg/kg/day) on steroidogenic effects of a relatively low-dose hCG (10 IU/rat, single dose, s.c.). Pretreatment of diabetic rats with TP03 enhanced the stimulatory effect of hCG on testosterone levels, slightly modifying its effects on the expression of steroidogenic proteins (Star, Cyp11a1, Cyp17a1) and LH/hCG receptor (Lhr) genes. Thus, in type 1 diabetes, TP03 increases the steroidogenic effect of low-dose hCG, at the same time as maintaining its effect on the gene expression of LH/hCG receptor and steroidogenesis enzymes in the testes.

Embryonic stem cells-derived mesenchymal stem cells do not differentiate into ovarian cells but improve ovarian function in POF mice

Premature ovarian failure (POF) is characterized by amenorrhea, hypoestrogenism, elevated gonadotropin levels, and infertility. Although some of previous studies reported that Mesenchymal stem cells (MSC) transplantation could rescue the ovary function of POF animal models through the paracrine pathways, these mechanisms require further investigation. Here, we aimed to investigate the possible mechanisms of therapeutic effects of human embryonic stem cells derived MSC (ES-MSC) in a mice model of chemotherapy-induced POF. For this purpose, Cyclophosphamide (Cy) was injected intraperitoneally into female mice to induce POF. 10 days after Cy injection, we evaluated follicle count, follicle-stimulating hormone (FSH) and estradiol (E2) hormone concentrations, and TUNEL assay. Then, ES-MSC was transplanted into mice and the expression of Anti-müllerian hormone (AMH) and apoptosis was evaluated in ovary. Results indicated that ES-MSC reduced apoptosis in the follicles and increased the expression of AMH protein in the ovary of POF mice. So, ES-MSC may inhibit the apoptosis of ovarian granulosa cells. Then, to investigate the potential mechanisms of therapeutic effects of ES-MSC and their fate in the ovary, MSC were labeled with green fluorescent protein (GFP) before transplantation. Immunofluorescence staining indicated that although GFP-labeled ES-MSC was located in the ovarian stroma, they did not express granulosa cell markers: AMH and Follicle-stimulating hormone receptor (FSHR), theca cell marker: luteinizing hormone receptor (LHR), and oocyte marker: Growth/differentiation factor 9 (GDF9). Therefore, ES-MSC may not differentiate into ovarian cells directly and they might restore ovarian function in chemotherapy-induced POF mice by paracrine mechanisms.

Role and mechanism of the p-JAK2/p-STAT3 signaling pathway in follicular development in PCOS rats

ObjectiveTo identify the association between the phosphorylated Janus kinase 2/phosphorylated signal transducer and activator of transcription (p-JAK2/p-STAT3) signaling pathway and follicular development in polycystic ovary syndrome (PCOS) rats, and explore the underlying mechanism. To evaluate the role of exogenous JAK2 inhibitor AG490 in the model and the associations among luteinizing hormone/choriogonadotropin receptor (LHCGR), follicle-stimulating hormone receptor (FSHR), cytochrome P450 17α (CYP17a), cytochrome P450 19 (CYP19), and PCOS. ResultsRat models of PCOS was established. PCOS rats were intraperitoneally treated with double-distilled water (ddH2O)/DMSO/AG490. The rate of ovarian morphological recovery in the AG490 group was significantly higher compared with the DMSO group (83.3 % vs 9.1 %, X2 = 12.68, P < 0.001). Moreover, the short in the time the estrous cycle was resumed in the AG490 group (hazard ratio = 16.32, P < 0.001) compared with the DMSO group. Compared with the controls, p-JAK2, p-STAT3, LHCGR, and CYP17a expression levels were increased whereas that of FSHR and CYP19 were decreased in the ovaries of PCOS rats. However, an opposite trend was observed after treatment with AG490. Software prediction revealed that the p-STAT3 bound to the promoter regions of LHCGR, FSHR, CYP17a, and CYP19 genes. This finding was confirmed by results of correlation analysis (R = 0.834, −0.836, 0.875 and −0.712, respectively, all P < 0.001). ConclusionThis study demonstrated that the p-JAK2/p-STAT3 signaling pathway was involved in follicular development in PCOS rats by upregulating LHCGR and CYP17a expression, and downregulating that of FSHR and CYP19. AG490 treatment exerted beneficial effects. LHCGR, FSHR, CYP17a, and CYP19 are candidate genes associated with follicular development in PCOS rats.

Effects of whole-body vibration on reproductive physiology in a rat model of whole-body vibration

ABSTRACT Findings from epidemiological studies suggest that occupational exposure to whole-body vibration (WBV) may increase the risk of miscarriage and contribute to a reduction in fertility rates in both men and women. However, workers exposed to WBV may also be exposed to other risk factors that contribute to reproductive dysfunction. The goal of this experiment was to examine the effects of WBV on reproductive physiology in a rat model. Male and female rats were exposed to WBV at the resonant frequency of the torso (31.5 Hz, 0.3 g amplitude) for 4 hr/day for 10 days. WBV exposure resulted in a significant reduction in number of developing follicles, and decrease in circulating estradiol concentrations, ovarian luteinizing hormone receptor protein levels, and marked changes in transcript levels for several factors involved in follicular development, cell cycle, and steroidogenesis. In males, WBV resulted in a significant reduction in spermatids and circulating prolactin levels, elevation in number of males having higher circulating testosterone concentrations, and marked alterations in levels of transcripts associated with oxidative stress, inflammation, and factors involved in regulating the cell cycle. Based upon these findings data indicate that occupational exposure to WBV contributes to adverse alterations in reproductive physiology in both genders that may lead to reduction in fertility.

TSH receptor antibodies (TRAb) – A potential new biomarker for endometriosis

ObjectivesThe prevalence of several autoimmune diseases, including thyroid dysfunction, has been reported to be increased in patients with endometriosis. Upregulated thyroid stimulation hormone (TSH) receptors in ectopic endometrium and elevated serum titers of TSH receptor antibodies (TRAb) IgG in endometriosis patients indicates an overlap in pathophysiology. However, cross-reactivity with other antibodies must be excluded. The objective of this study was to compare the expression of autoantibodies in women with endometriosis and two control groups to evaluate the potential of TRAb IgG as a diagnostic marker for endometriosis. Study designThis cross-sectional study was carried out in 172 women with surgically confirmed endometriosis and two control groups consisting of 50 healthy blood donors and 114 women from Malmö Offspring Study consisting of people from the general population. Serum levels of thyroid hormones, TSH and TRAb autoantibodies, AXIN1, and autoantibodies against follicle stimulating hormone (FSH), human chorionic gonadotropin (hCG), luteinizing hormone (LH), and their receptors, were analyzed. The patients answered a questionnaire and estimated their gastrointestinal symptoms using the Visual Analogue Scale for Irritable Bowel Syndrome. ResultsOf the endometriosis patients, 29.1 % had TRAb IgG above the present detection limit of ≥ 1.0 IE/L compared to 2.6 % of the controls from MOS (p < 0.001) and 94.5 % had levels of TRAb over the previous detection limit ≥ 0.3 IE/L compared to 7.9 % of the controls (p < 0.001). Titers of both TRAb IgG and IgM were increased in patients compared to controls from MOS and blood donors, respectively (p < 0.001). There was no increase of autoantibodies against FSH, FSH receptor (FSHR), hCG, LH, LH receptor (LHR) or TSH compared to the blood donor controls. TRAb titers did not correlate with age, disease duration, AXIN1, TSH, thyroid hormones or gastrointestinal symptoms. ConclusionTRAb IgG and IgM are slightly elevated in patients with endometriosis with no cross-reactivity with other autoantibodies. The results indicate that TRAb is truly elevated and thereby has the potential to be used to support the diagnosing of endometriosis.

Effects of dietary organic acids and probiotics on laying performance, egg quality, serum antioxidants and expressions of reproductive genes of laying ducks in the late phase of production

Five hundred and forty Cheery Valley ducks were used to investigate the effects of dietary supplementation of acidifier and compound probiotics, individually or in combination, on production performance, egg quality, immune and oxidative status, expression of reproductive, and calcium binding related genes from 42 wk to 48 wk of age. Ducks were randomly allocated to 9 treatment groups with 6 replicates and 10 ducks per replicate for each group. A 3 × 3 factorial arrangement, with 3 dietary inclusion levels of acidifier and probiotics (0, 2, and 3 g/kg acidifier; 0, 1, and 2 g/kg probiotics) were used. The acidifier used was mainly consisted of Benzoic acid, Fumaric acid, phosphoric acid, and formic acid. The main components of the probiotics were Bacillus subtilis and Clostridium butyricum. Dietary supplementation of probiotics improved the daily feed intake, egg production rate, and body weight of ducks (P < 0.05), and diet acidifier also increased the daily feed intake compared to the control (P < 0.01). Egg quality was improved by diet inclusion of probiotics, including Haugh unit, albumen height, egg shape index (P < 0.01), and eggshell hardness (P = 0.05). A significant increase in Haught unit and yolk weight was observed in ducks fed diet added with acidifier (P < 0.05). Acidifier supplementation reduced the total antioxidant capacity (T-AOC), immunoglobulin A (IgA), and IgG content and the catalase (CAT) activity in the serum (P < 0.05), in accompanied with an increased malondialdehyde (MDA) concentration (P < 0.05). Serum total superoxide dismutase (T-SOD) activities were improved by dietary inclusion of probiotics (P < 0.05). There was an interaction effects on serum IgA and IgG contents between acidifier and probiotics (P < 0.05). Diet supplementation of probiotics improved the ovary follicle-stimulating hormone receptor (FSHR) and estrogen receptor (ER) gene expressions (P < 0.01), while dietary acidifier reduced the transcription levels of FSHR and luteinizing hormone receptor (LHR) (P < 0.01) in ovary. In the uterus of the oviduct, expressions of FSHR, and carbonic anhydrase 2 (CA2) were also increased by diet probiotics (P < 0.01), and diet acidifier reduced the gene expressions of calbindin-D28k (CaBP-D28k) and CA2 (P < 0.05). Significant interaction effects between diet acidifier and probiotics were obtained on gene expressions of FSHR, LHR, and ovalbumin (OVAL) in the ovary (P < 0.05), and LHR, CaBP-D28k, and CA2 (P < 0.05) in the uterus. It can be concluded that production performance and egg quality of laying ducks can be improved in the late phase of reproduction by dietary inclusion of probiotics, while the organic acid mixture caused a decline in serum antioxidant and immune capacity of the ducks.

Effects of bisphenol AF on growth, behavior, histology and gene expression in marine medaka (Oryzias melastigma)

Bisphenol AF (BPAF) is one of the substitutes for bisphenol A (BPA), which has endocrine-disrupting, reproductive and neurological toxicity. BPAF has frequently been detected in the aquatic environment, which has been a long-term threat to the health of aquatic organisms. In this study, female marine medaka (Oryzias melastigma) were exposed to 6.7 μg/L, 73.4 μg/L, and 367.0 μg/L BPAF for 120 d. The effects of BPAF on behavior, growth, liver and ovarian histology, gene transcriptional profiles, and reproduction of marine medaka were determined. The results showed that with the increase of BPAF concentration, the swimming speed of female marine medaka showed an increasing trend and then decreasing trend. BPAF (367.0 μg/L) significantly increased body weight and condition factors in females. BPAF (73.4 μg/L and 367.0 μg/L) significantly delayed oocyte maturation. Exposure to 367.0 μg/L BPAF showed an increasing trend in the transcript levels of lipid synthesis and transport-related genes such as fatty acid synthase (fasn), sterol regulatory element binding protein (srebf), diacylglycerol acyltransferase (dgat), solute carrier family 27 member 4 (slc27a4), fatty acid-binding protein (fabp), and peroxisome proliferator-activated receptor gamma (pparγ) in the liver. In addition, 6.7 μg/L BPAF significantly down-regulated the expression levels of antioxidant-related genes [superoxide dismutase (sod), glutathione peroxidase (gpx), and catalase (cat)], and complement system-related genes [complement component 5 (c5), complement component 7a (c7a), mannan-binding lectin serine peptidase 1 (masp1), and tumor necrosis factor (tnf)] were significantly up-regulated in the 73.4 and 367.0 μg/L groups, which implies the effect of BPAF on the immune system in the liver. In the hypothalamic-pituitary-ovarian axis (HPG) results, the transcription levels of estrogen receptor α (erα), estrogen receptor β (erβ), androgen receptor (arα), gonadotropin-releasing hormone 2 (gnrh2), cytochrome P450 19b (cyp19b), aromatase (cyp19a), and luteinizing hormone receptor (lhr) in the brain and ovary, and vitellogenin (vtg) and choriogenin (chg) in the liver of 367.0 μg/L BPAF group showed a downward trend. In addition, exposure to 367.0 μg/L BPAF for 120 d inhibited the spawning behavior of marine medaka. Our results showed that long-term BPAF treatment influenced growth (body weight and condition factors), lipid metabolism, and ovarian maturation, and significantly altered the immune response and the transcriptional expression levels of HPG axis-related genes.

Attainment of Sexual Maturity and Gonadotropin Priming in Gilts Determine Follicular Development, Endocrine Milieu and Response to Ovulatory Triggers.

The routine procedure of estrous cycle synchronization in pigs allows for the use of gonadotropins to stimulate ovarian activity. The applied protocols of eCG and hFSH priming similarly affected development of ovarian follicles in two classes 3–6 mm and >6 mm of diameter, however, the number of small follicles (<3 mm) was 2-fold higher in hFSH- than in eCG-primed prepubertal gilts. The attainment of sexual maturity increased concentration of estradiol, testosterone and androstenedione in the follicular fluid of hFSH/eCG-primed gilts, however, prostaglandin E2 and F2α metabolite increased in mature hFSH- and eCG-primed gilts, respectively. The maturity increased mRNA and/or protein expression of key steroidogenic enzymes, prostaglandin synthases or luteinizing hormone receptors in follicular walls. Both hormonal primers played a moderate role in affecting expression of steroidogenic enzymes in follicular walls. In vitro studies showed higher estradiol production in r-hLH (p = 0.04)- and r-hCG (p = 0.049)-stimulated follicular walls of mature gilts than in prepubertal hFSH-primed gilts. Both ovulatory triggers decreased the abundance of LHCG/FSH mRNA receptors in follicular walls, which mimic downregulation of these receptors by a preovulatory LH surge, confirmed in vivo. These data revealed the importance of sexual maturity in the protection of the estrogenic environment, and the selective, moderate role of eCG and FSH in the activation of steroidogenic enzymes in preovulatory follicles.

Aerobic Training and Crocin Consumption Improve Doxorubicin Damage to Spermatogenesis Process and Luteinizing Hormone Receptor Expression of Male Wistar Rats

Background: Many men in the world are infertile at reproductive age. One of the causes of male infertility is the side effects of chemotherapy drugs. Doxorubicin (DOX) is a tetracycline used in chemotherapy that has been shown to have adverse effects on testicular tissue. Objectives: The simultaneous effect of high-intensity aerobic exercise and crocin (Cr) consumption on doxorubicin-induced testicular tissue has not been studied. Therefore, the aim of this study was to investigate the interaction between exercise and Cr consumption on spermatogenesis and expression of lutein hormone receptor (LHR) in testicular tissue of rats injected with doxorubicin. Methods: Thirty male Wistar rats were randomly divided into five groups (n = 6), including (1) healthy control, (2) doxorubicin, (3) doxorubicin + 10 mg/kg body weight (kg.bw) Cr, (4) doxorubicin + high intensity interval training (HIIT) and (5) doxorubicin + aerobic training with 10 mg/kg.bw Cr. Rats in groups 3 and 5 received Cr daily at specific doses orally, and groups 4 and 5 performed aerobic training (2 - 8 rep × 2 min at 80 - 110% Vo2max) five days/w. Also, groups 2 to 5 received two mg/kg of doxorubicin peritoneally seven times throughout the study. Expression of LHR and spermatogenesis was measured by immunohistochemistry (IHC) and hematoxylin and eosin (H&amp;E), respectively. Results: Data analysis showed a significant decrement in the spermatogenesis and expression of LHR in the doxorubicin group compared with the control group (P ≤ 0.0001). Also, the interaction of HIIT and Cr caused the most significant increase in LHR expression and spermatogenesis (P ≤ 0.03). The effect of HIIT on LHR expression and spermatogenesis was higher than that of Cr consumption (P ≤ 0.05). Conclusions: To prevent the harmful side effect of doxorubicin on testicular tissue HIIT and Cr consumption are recommended. Also, the positive effect of HIIT was significantly higher than Cr consumption.