Expression Levels Of Follicle-stimulating Hormone Receptor Research Articles

Unkeito promotes follicle development by restoring reduced follicle-stimulating hormone responsiveness in rats with polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) is a common disorder resulting in irregular menstruation and infertility due to improper follicular development and ovulation. PCOS pathogenesis is mediated by downregulated follicle-stimulating hormone receptor (FSHR) expression in granulosa cells (GCs); however, the underlying mechanism remains elusive. Unkeito (UKT) is a traditional Japanese medicine used to treat irregular menstruation in patients with PCOS. In this study, we aimed to confirm the effectiveness of UKT in PCOS by focusing on follicle-stimulating hormone (FSH) responsiveness. A rat model of PCOS was generated by prenatal treatment with 5α-dihydrotestosterone. Female offspring (3-week-old) rats were fed a UKT mixed diet or a normal diet daily. To compare the PCOS phenotype in rats, the estrous cycle, hormone profiles, and ovarian morphology were evaluated. To further examine the role of FSH, molecular, genetic, and immunohistological analyses were performed using ovarian tissues and primary cultured GCs from normal and PCOS model rats. UKT increased the number of antral and preovulatory follicles and restored the irregular estrous cycle in PCOS rats. The gene expression levels of FSHR and bone morphogenetic protein (BMP)-2 and BMP-6 were significantly decreased in the ovarian GCs of PCOS rats compared to those in normal rats. UKT treatment increased FSHR staining in the small antral follicles and upregulated Fshr and Bmps expression in the ovary and GCs of PCOS rats. There was no change in serum gonadotropin levels. In primary cultured GCs stimulated by FSH, UKT enhanced estradiol production, accompanied by increased intracellular cyclic adenosine monophosphate levels, and upregulated the expression of genes encoding the enzymes involved in local estradiol synthesis, namely Cyp19a1 and Hsd17b. Furthermore, UKT elevated the expression of Star and Cyp11a1, involved in progesterone production in cultured GCs in the presence of FSH. UKT stimulates ovarian follicle development by potentiating FSH responsiveness by upregulating BMP-2 and BMP-6 expression, resulting in the recovery of estrous cycle abnormalities in PCOS rats. Restoring the FSHR dysfunction in the small antral follicles may alleviate the PCOS phenotype.

Changes in histology, protein expression, and autophagy in dairy goat testes during nonbreeding season†.

Seasonal reproduction contributes to increased chances of offspring survival in some animals. Dairy goats are seasonal breeding mammals. In this study, adult male Guanzhong dairy goats (10-12months old) were used. Testis size, semen quality, hormone level, apoptosis of germ cells, and autophagy of Sertoli cells were analyzed in dairy goats during the breeding (October) and nonbreeding (April) seasons. We found that, during the nonbreeding season for dairy goats, semen quality, follicle-stimulating hormone (FSH) levels, and testosterone levels were reduced, and the number of apoptotic germ cells increased. The proliferation with decrease activity of germ cells in dairy goat during the nonbreeding season was significantly affected. However, the testis size did not change seasonally. Interestingly, Sertoli cell autophagy was more active during the nonbreeding season. The expression levels of FSH receptor, wilms tumor 1, androgen binding protein, glial cell derived neurotrophic factor, and stem cell factor decreased in dairy goats during the nonbreeding season. In summary, our results indicate that spermatogenesis in dairy goats during the nonbreeding season was not completely arrested. In addition, germ cell apoptosis and the morphology of Sertoli cells considerably changed in dairy goats during the nonbreeding season. Sertoli cell autophagy is involved in the seasonal regulation of spermatogenesis in dairy goats. These findings provide key insights into the fertility and spermatogenesis of seasonal breeding animals.

Expression levels of FSHR, IGF1R, CYP11al and HSD3β in cumulus cells can predict in vitro developmental competence of bovine oocytes.

The efficiency of in vitro embryo production technologies would be improved by the development of suitable non-invasive biomarkers that allow the selection of good quality cumulus-oocyte complexes (COCs). The present study used whole, single oocyte culture to investigate whether the expression levels of follicle-stimulating hormone receptor (FSHR), insulin-like factor 1 receptor (IGF1R) and three steroidogenesis-related enzymes (CYP11al, CYP19al and HSD3β) in cumulus cells reflected the developmental competence of COCs. Cumulus cells were collected from single COCs before maturation culture and relative mRNA levels were assessed using real-time PCR. The analysis indicated that mRNAs for FSHR, IGF1R, CYP11al and HSD3β were present at higher levels in cumulus cells from COCs that failed to form blastocysts compared with cumulus cells from COCs that formed blastocysts. Moreover, FSHR and IGF1R mRNA levels were positively correlated with those of genes for steroidogenesis-related enzymes. In conclusion, poor developmental competence of COCs was related to higher expression of FSHR, IGF1R, CYP11al and HSD3β in cumulus cells, which may indicate the advanced differentiation of cumulus cells into granulosa cells.

Growth differentiation factor 9 promotes follicle-stimulating hormone-induced progesterone production in chicken follicular granulosa cells

The function of oocyte-derived growth differentiation factor 9 (GDF9) in ovarian follicles has thus far been poorly defined in avian species compared with the defined function in mammals. Our aim here is to investigate the effects of GDF9 on steroidogenesis and on chicken ovarian granulosa cell (GC) mitosis. Primary GCs from both prehierarchical (6–8 mm in diameter, phGCs) and preovulatory follicles (F1–F5, poGCs) were cultured in the presence or absence of the GDF9 protein. The progesterone (P4) levels in the culture medium were then measured by radioimmunoassay (RIA), and the expression levels of steroidogenesis genes were detected by quantitative PCR. We found that GDF9 alone showed no significant effect on the P4 levels by regulating the expression of steroidogenesis genes, such as STAR, CYP11A1 and HSD3B. Further experiments indicated that GDF9 promoted follicle-stimulating hormone (FSH)-induced P4 production and STAR expression. GDF9 also rescued the FSH-induced decrease of FSH receptor (FSHR) expression but had no effect on the forskolin-induced P4, STAR and forskolin-inhibited FSHR expression levels, suggesting that GDF9 might achieve its regulatory role of P4 by enhancing FSHR and STAR expression. In addition, GDF9 also promoted GC cell cycle progression, regulated the gene transcription of related genes, potentiated DNA replication and inhibited apoptosis. Interestingly, these effects differed between the phGCs and the poGCs. To our knowledge, this is the first report that illustrates the function of GDF9 on chicken GCs and the effects on ovarian steroidogenesis. Our findings highlight the regulation of central oocytes on the surrounding granulosa cells and emphasize the interaction between paracrine signals and endocrine hormones on ovarian progesterone production; these findings contribute to the understanding of the development of avian ovarian follicles.

MiR-31 and miR-143 affect steroid hormone synthesis and inhibit cell apoptosis in bovine granulosa cells through FSHR

The regulatory role of microRNAs (miRNAs) has been explored in ovarian cells, and the effects of miRNAs on gonadal development, apoptosis, ovulation, and steroid production have been reported. In this study, we analyzed the effects of follicle stimulating hormone (FSH) on miR-31 and miR-143 expression levels in bovine granulosa cells (GCs). Our results demonstrated that the FSH receptor (FSHR) is a common target gene of miR-31 and miR-143 in bovine GCs. We further analyzed the roles of miR-31 and miR-143 in bovine GCs by transfecting miR-31 and miR-143 mimics and inhibitors. The Western blot and RT-PCR results showed that miR-31 and miR-143 reduced the mRNA and protein expression levels of FSHR. Moreover, miR-31 overexpression decreased the secretion of progesterone (P4), and miR-143 overexpression decreased both the synthesis of P4 and the secretion of estrogen (E2). In contrast, miR-31 inhibition increased the secretion of progesterone (P4), and miR-143 inhibition increased both the synthesis of P4 and the secretion of E2. Finally, we analyzed the possible effects of miR-31 and miR-143 on bovine GC apoptosis. The results showed that transfection with miR-31 and miR-143 mimics promoted GC apoptosis and that miR-143 and miR-31 inhibition reduced the rate of apoptosis in bovine GCs. Taken together, our results indicate that miR-31 and miR-143 decrease steroid hormone synthesis and inhibit bovine GC apoptosis by targeting FSHR.

Effects of Thyroid Dysfunction on Reproductive Hormones in Female Rats.

Thyroid hormones (THs) play a critical role in the development of ovarian cells. Although the effects of THs on female reproduction are of great interest, the mechanism remains unclear. We investigated the effects of TH dysregulation on reproductive hormones in rats. Propylthiouracil (PTU) and L-thyroxine were administered to rats to induce hypo- and hyperthyroidism, respectively, and the reproductive hormone profiles were analyzed by radioimmunoassay (RIA). Ovarian histology was evaluated with hematoxylin and eosin (H&E) staining, and gene protein level or mRNA content was analyzed by western blotting or reverse transcription polymerase chain reaction (RT-PCR). The serum levels of gonadotropin releasing hormone (GnRH) and follicle stimulating hormone (FSH) in both rat models were significantly decreased on day 21, although there were no significant changes at earlier time points. There were no significant differences in luteinizing hormone (LH) or progesterone (P4) levels between the treatment and the control groups. Both PTU and L-thyroxine treatments downregulated estradiol (E2) concentrations; however, the serum testosterone (T) level was increased only in hypothyroid rats at day 21. In addition, the expression levels of FSH receptor, cholesterol side-chain cleavage enzyme (P450scc), and steroidogenic acute regulatory protein (StAR) were decreased in both rat models. Moreover, the onset of puberty was significantly delayed in the hypothyroid group. These results provide evidence that TH dysregulation alters reproductive hormone profiles, and that the initiation of the estrous cycle is postponed in hypothyroidism.

Seasonal expressions of follicle-stimulating hormone receptor and luteinizing hormone receptor in the scented gland of the male muskrat (Ondatra zibethicus).

Accumulating evidence has shown that follicle-stimulating hormone (FSH) and luteinizing hormone (LH) may influence the functions of nongonadal tissues in addition to their classic target gonads. Our previous studies revealed that the scented glands of male muskrats expressed prolactin receptor, steroidogenic enzymes, and inhibin/activin subunits. To further seek the evidence of the activities of pituitary gonadotropins in scented glands, we investigated the seasonal expression patterns of FSH receptor (FSHR) and LH/choriogonadotropin receptor (LHCGR). The weight and size of scented glands during the breeding season were significantly higher than those during the nonbreeding season. Immunohistochemical studies showed that FSHR was present in the serous cells of scented glands, whereas LHCGR was present in the interstitial cells. The protein and mRNA expression levels of FSHR and LHCGR were significantly higher in the scented glands during the breeding season than those during the nonbreeding season. Importantly, the levels of circulating FSH and LH were remarkably higher during the breeding season. Taken together, these results suggested that gonadotropins may affect the function of muskrat scented gland via the locally expressed receptors in a season-dependent manner.

Maturation rates of oocytes and levels of FSHR, LHR and GnRHR of COCs response to FSH concentrations in IVM media for sheep

This study investigated the FSH influence on maturation rates of oocytes in vitro maturation (IVM), and expression levels of the follicle-stimulating hormone receptor (FSHR), luteinizing hormone receptor (LHR) and gonadotropin releasing hormone receptor (GnRHR) of cumulus-oocyte complexes (COCs) response to FSH treatment. 1686 COCs were harvested from 1063 ovaries of sheep. COCs were cultured 26h at 38.5°C and 5.0% CO2 in IVM media supplemented with 0, 5, 10, 20 and 30 IU/ml FSH. They were allocated in to FSH-1 (basal line), FSH-2, FSH-3, FSH-4 and FSH-5 groups. The apoptosis of COCs was assessed by Tunel assay. Expression levels of mRNA and protein for FSHR, LHR and GnRHR in sheep COCs were detected using real time RT-PCR and Western blotting respectively. The results showed that the maturation rates of oocytes were improved gradually when FSH supplement increased from 0 to 10μg/ml. FSH-3 group showed the highest maturation rate. Apoptosis rates of FSH-treated groups were less than that of FSH-1 group with a minimum of FSH-3 group. Expression levels of FSHR and LHR mRNAs in FSH-3 and FSH-4 were significantly higher than in FSH-1. Expression level of GnRHR mRNA in FSH-3 was higher than in FSH-1 (P<0.05). Expression levels of FSHR proteins in FSH-3 and FSH-4 groups were higher than that of FSH-1 group. Expression levels of GnRHR proteins increased gradually with a maximal increment of FSH-5. Maturation rates of COCs had significant positive correlations with mRNA and protein levels of FSHR, LHR and GnRHR. In conclusion, FSH could accelerate the maturation rate of sheep oocytes and reduce their apoptosis rate, also increase the expression levels of FSHR, LHR and GnRHR mRNAs, and strengthen expressions of FSHR and GnRHR proteins. 10 IU/ml FSH additions were the optimal dose for IVM of sheep oocytes.

Cetrorelix and Triptorelin active immunization influences follicle development and receptor expressions of ovaries in mice

The study investigated comparatively the effects of active immunization of Cetrorelix and Triptorelin on the follicle development, histological structures, and expressions of FSHR and LHR proteins in the ovary. One hundred and five female mice, 35 days old, and body weight of 26.56±2.89g, were randomly assigned into Cetrorelix (CET), Triptorelin (TRI) and control groups (CG). Mice in CET-1, CET-2 and CET-3 (n=15) were subcutaneously injected with 0.1, 0.2 and 0.4mL Cetrorelix antigen (100μg/mL) for seven days, respectively. Mice in TRI-1, TRI-2 and TRI-3 were injected with 0.1, 0.2 and 0.4mL Triptorelin antigen (100μg/mL) for seven days. Mice in CG (n=15) were injected with 0.2mL saline for seven days. Western blotting was utilized to determine the expressions of follicle stimulating hormone receptor (FSHR) and luteinizing hormone receptor (LHR) proteins. ELISA was used to measure the serum FSH and LH concentrations. The ovarian slices were observed under optical microscopy. The results showed that the ovarian weights of CET and TRI groups increased slightly with a maximum increment of 27.02% of CET-1 mice on day 35. Ovarian weights in TRI group increased dose-dependently with a maximum increment of 45.77% in TRI-3 on day 35. The numbers of the primordial follicle (POF), primary follicle (PF), secondary follicle (SF) and mature follicle (MF) of CET and TRI groups increased at different degrees when compared to CG. The granular layer in SF was arranged tightly and zona pellucida (ZP) was thickened. Follicles developed fully. Follicle longitudinal diameter (FLD), follicle transverse diameter (FTD) and follicle wall thickness (FWT) in CET and TRI groups increased as compared to CG. FLD, FTD and FWT of TRI-3 had a larger increment than CG and CET-1 on days 21 and 35. Expression levels of FSHR and LHR proteins in TRI group increased when compared to CG. Expression levels of FSHR and LHR proteins of CET group decreased. Serum FSH of TRI group was slightly higher than the CG and CET group on day 21. On day 35, serum FSH levels of CET-1 and TRI-1 were greater than CG and the rest of the subgroups. Serum FSH levels had no significant differences among all groups. Expression levels of LHR and FSHR proteins in CET group had obvious negative correlations to FLD, FTD and FWT. However, in the TRI group, these correlations were positive. In conclusion, Cetrorelix and Triptorelin immunization could improve ovarian growth, increase the follicle numbers, enhance dose-dependently the FLD, FTD and FWT, and eventually promote the ovary and follicle development. Cetrorelix decreased expression levels of FSHR and LHR proteins in the ovaries of mice. Triptorelin enhanced expression levels of FSHR and LHR proteins. Triptorelin treatment had more obvious effects than Cetrorelix treatment.

The extracts of Pacific oyster (Crassostrea gigas) alleviate ovarian functional disorders of female rats with exposure to bisphenol a through decreasing FSHR expression in ovarian tissues.

Bisphenol-A (BPA) is one of the widespread industrial compounds, which has adverse effects on animal and human health. The study was aimed to explore the effects of Crassostrea gigas extracts (CGE) in alleviating ovarian functional disorders of female rats with exposure to BPA and the underlying possible mechanism. Eighteen four-week-old female Sprague-Dawley (SD) rats were randomly divided into BPA group (50mg/kg BPA), BPA+CGE group (50mg/kg BPA+50mg/kg CGE), and control group (equivalent dosage of vehicle) with 6 rats in each group. After a 6-week treatment ended, the serum levels of estradiol (E2), follicle stimulating hormone (FSH), luteinizing hormone (LH) were measured by using commercial standard assay kits. The expression levels of FSH receptor (FSHR) in the rat ovarian tissues were respectively detected by immunohistochemistry and Real-time PCR. CGE treatment markedly increased E2 levels and decreased FSH levels in the serum (P<0.05), however, the alterations of serum LH levels were not significant (P>0.05). The protein and mRNA expression levels of FSHR were the lowest in the ovaries of control rats and the highest in BPA rats (P<0.05). CGE treatment markedly decreased the expression levels of FSHR in the ovarian tissues (P<0.05). Crassostrea gigas successfully alleviates ovarian functional disorders of female rats with exposure to BPA partly through decreasing FSHR expression levels in the ovarian tissues.

Potential detrimental effect of soy isoflavones on testis sertoli cells

To determine the effect of soy isoflavones on cell proliferation and the transcription levels of follicle-stimulating hormone receptor (FSHR), inhibin α (INHα), INHβB, androgen binding protein (ABP), transferrin (Tf) and vimentin in testis sertoli cells in SD rats. Sertoli cells were cultured in vitro, exposed to daidzein at 0.03, 0.3, 3, and 30 μmol/L and genistein at 0.05, 0.5, 5 and 50 μmol/L, respectively. MTT was used to detect the proliferation of sertoli cells. Real-time PCR was used to detect the relative mRNA expressions of FSHR, INHα, INHβB, ABP, Tf and vimentin. Compared with control groups, cell proliferation and the relative mRNA expression levels of INHβB and ABP in the treated cells showed no significant alternation. The INHα mRNA expression levels were increased in 0.3 and 3 μmol/L Dai and 0.05 μmol/L Gen, while the mRNA expression levels of FSHR were downregulated in 30 μmol/L Dai and Gen at all concentrations. Tf mRNA expression levels were downregulated in 30 μmol/L Dai and 5 μmol/L and 50 μmol/L Gen, and the mRNA expression levels of vimentin were downregulated in 3 and 30 μmol/L Dai and 50 μmol/L Gen. Soy Isoflavones may have potential detrimental effect on the male reproductive system, as they may impact the function of sertoli cells by downregulating the transcription levels of some important proteins.

Endothelial follicle stimulating hormone receptor in primary kidney cancer correlates with subsequent response to sunitinib

Sunitinib is an anti-angiogenic receptor tyrosine kinase inhibitor used to treat advanced metastatic renal cell carcinoma and other types of cancer. Sutent is effective in only approximately 70% of clear cell renal cell carcinoma (CCRCC) patients, has significant adverse side effects and no method is available to predict which patients will not respond. Our purpose was to explore the possibility of introducing an effective prediction method based on a marker of the tumour vasculature, the follicle stimulating hormone receptor (FSHR). Fifty patients diagnosed with advanced metastatic CCRCC have been subjected to surgery for removal of the primary tumour and were subsequently treated with sunitinib. After three months of therapy the patients were categorized as ‘responsive’, ‘stable’ or ‘non-responsive’ based on the RECIST guidelines. The blood vessel density and the percentage of FSHR-positive vessels were determined by immunofluorescence on sections from the primary tumours removed by surgery, prior to the sunitinib treatment. The percentage of FSHR-stained vessels was on average fivefold higher for the patients who responded to the treatment in comparison with the stable group and almost eightfold higher than in the non-responsive group. The percentage allowed the detection of responders with 87–100% sensitivity and specificity. No significant differences were detected in the total density of vessels among the three groups. The data suggest that FSHR expression levels in the blood vessels of CCRCC primary tumours can be used to predict, with high sensitivity and specificity, the patients who will respond to sunitinib therapy.

The level of FSHR expression in the primary kidney tumors to predict the response to subsequent antiangiogenic therapy with sunitinib.

e21138 Background: Sunitinib is an antiangiogenic receptor tyrosine kinase inhibitor used to treat advanced metastatic renal cell carcinoma and other types of cancer. Sunitinib is effective in only approx. 70% of clear cell renal cell carcinoma (CCRCC) patients, has significant adverse side effects, and no method is available to predict which patients will not respond. Our purpose was to explore the possibility of introducing an effective prediction method based on a marker of the tumor vasculature, the Follicle Stimulating Hormone Receptor (FSHR). Methods: Using immunohistochemistry on paraffin sections with anti-FSHR monoclonal antibodies we studied by photonic and fluorescence microscopy the expression of FSHR in 50 patients diagnosed with advanced metastatic CCRCC. All patients were subjected to surgery for removal of the primary tumor and were subsequently treated with sunitinib orally for ≥ 3 months with a dose of 50 mg/day for 4 weeks followed by 2 weeks off. After therapy the patients were categorized as “responsive”, “stable”, or “non-responsive” based on the RECIST guidelines. The blood vessel density and the percentage of FSHR-positive vessels were determined by immunofluorescence on sections from the primary tumors removed by surgery, prior to the sunitinib treatment. Von Willebrand factor (vWF) was used as specific endothelial marker. Results: There were no significant differences in the total density of vessels (detected with anti-vWF antibody) among the responsive, stable and non-responsive patients. The percentage of FSHR-stained vessels was on average five fold higher for the patients that responded to the treatment in comparison with the stable group and almost eight fold higher than in the non-responsive group. The percentage allowed the detection of responders with 87-100% sensitivity and specificity. Conclusions: The data suggest that FSHR expression levels in the blood vessels of CCRCC primary tumors can be used to predict, with high sensitivity and specificity, the patients that will respond to sunitinib therapy.

Molecular cloning of two gonadotropin receptors and their distinct mRNA expression profiles in daily oogenesis of the wrasse Pseudolabrus sieboldi

In fish, asynchronous development of ovarian follicles, the simultaneous advance of vitellogenesis and oocyte maturation in one ovary, is a rational reproductive strategy to spawn consecutively in one spawning season. In this study, to clarify the mode of action of follicle stimulating hormone (FSH) and luteinizing hormone (LH) in asynchronous ovarian follicle development in daily egg production, we cloned cDNAs of the follicle-stimulating hormone receptor (FSHR) and luteinizing hormone receptor (LHR) in the bambooleaf wrasse ( Pseudolabrus sieboldi), which exhibits clear diurnal spawning rhythms over 1 month. In addition, different developmental stages of ovarian follicles were isolated from whole ovaries at various daily time points on 1 day in the spawning season, and mRNA expression levels of FSHR and LHR were analyzed. Sequence analysis showed distinct differences in the number of putative leucine-rich repeats at the extracellular domain between FSHR and LHR, suggesting a difference in ligand-specificity. Real-time PCR analyses revealed that FSHR mRNA was highly expressed in early yolk-stage follicles but decreased at the end of vitellogenesis. In contrast, the expression of LHR mRNA was maintained at low levels in vitellogenic stage follicles but markedly elevated at the end of the vitellogenic and early migratory nucleus stages, thereafter markedly dropping in the late migratory nucleus stage. The present results suggest that co-regulation of vitellogenesis and oocyte maturation in one ovary is controlled by the stage-distinctive expression levels of FSHR and LHR mRNA in ovarian follicles, and daily switching of sensitivity from FSH to LH is required for daily egg production.