Follicle-stimulating Hormone Activity Research Articles

Health risk and germ cell toxicity of five commercially available sachet waters in Nigeria: a public health concern.

Sachet water is the most common form of portable water commercially available in Nigeria. Using the murine sperm count and sperm abnormality assay, the germ cell toxicity of five common commercially available sachet waters in Nigeria was assessed in this study. The levels of hormones such as Follicle Stimulating Hormone (FSH), Luteinizing Hormone (LH) and Total Testosterone (TT); and activities of catalase (CAT), alanine aminotransferase (ALT), superoxide dismutase (SOD), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) were evaluated. The heavy metal and physicochemical parameters of the sachet waters were also analyzed. Healthy male mice were allowed to freely drink the sachet waters for 35days after which they were sacrificed. The findings indicated that the concentrations of some heavy metals (As, Cr, and Cd) in the sachet waters exceeded the limit by regulatory organizations. The data of the total carcinogenic risk (TCR) and total non-carcinogenic risk (THQ) of some heavy metals associated with the ingestion of sachet water for adults and children showed that the values exceeded the acceptable threshold, and thus, is indicative of a high non-carcinogenic and carcinogenic risks. The data of the sperm abnormality assay showed that in the exposed mice, the five sachet waters induced a statistically significant (P<0.05) increase in abnormal sperm cells and a significantly lower mean sperm count. Additionally noted were changes in the serum activities of TT, FSH, ALP, AST, ALT, and LH. Thus, the sachet waters studied contained agents that can induce reproductive toxicity in exposed humans. This is of public health importance and calls for immediate action by regulatory bodies.

An Array SPRi Biosensor for the Determination of Follicle-Stimulating Hormone in Blood Plasma.

Follicle-stimulating hormone (FSH) regulates the development, growth, pubertal maturation and reproductive processes of the human body. The determination of serous FSH concentration is significant as an alternative to testicular biopsy in the case of boys suffering from cryptorchidism after orchidopexy, and as a means of determining the menopausal stage in women. The aim of this investigation is to develop a specific array surface plasmon resonance imaging (SPRi) biosensor for the determination of FSH in body liquids such as blood plasma, obtaining sufficient sensitivity to determine FSH at levels characteristic for that hormone in blood plasma, without any signal enhancement. The biosensor consists of a mouse monoclonal anti-FSH antibody attached to the gold surface of a chip via a cysteamine linker. Its linear response range is from 0.08 mIU mL-1 (LOQ) to 20 mIU mL-1, and well covers most of the range of FSH activities found in blood without dilution. The precision of measurement is between 3.2% and 13.1% for model samples, and between 3.7% and 5.6% for spiked plasma samples. Recoveries are in the range from 94% to 108%. The biosensor has good selectivity, and is validated by comparison with ECLE, with good agreement of the results.

Determination of biological activity of gonadotrophins in inbred and outbred animals. Part 1: Determination of biological activity of follicle-stimulating hormone

Scientific relevance. The biological activity of medicinal products may vary depending on the method of production (i.e. biological or recombinant products). The widening variety of gonadotrophin preparations, the diversity of their production methods, and the irreplaceability of biological activity bioassays with physicochemical tests require improvement of animal testing conditions.Aim. This study aimed to determine the biological activity of follicle-stimulating hormone (FSH) in several rat lines, analyse the findings, and select the most optimal testing conditions.Materials and methods. The biological activity was determined using in vivo methods. The comparative analysis used test results obtained over several years in inbred and outbred rats treated with FSH. In all cases, the authors used a three-dose randomised method based on the determination of the biological activity of test samples by comparison with that of the WHO international standard (IS) containing 183 IU of FSH bioactivity and 177 IU of LH bioactivity per ampoule (NIBSC code: 10/286). The study included immature female rats, inbred (Wistar-Kyoto or Sprague Dawley) and outbred. Testing conditions depended on the selected rat line, with the main variables being the test dose and the number of animals per group.Results. The authors compared responses of inbred and outbred rats to various doses of the FSH IS and test samples. Given the narrow range of the analytical procedure, Wistar-Kyoto rats showed a relatively weak dose–response relationship. The study demonstrated that the doses and testing duration depended on the sensitivity of the animals. Test results were less variable in inbred rats than in outbred ones. The statistical analysis of the results of FSH bioactivity testing in inbred and outbred rats showed that, with inbred rats, the number of animals could be halved without compromising the validity of the test.Conclusions. The approach proposed in this study provides for testing the biological activity of FSH with fewer experimental animals, improved cost-effectiveness, and the same reliability of results.

The Microanatomical Study of Effect of Alkaloid Isolate of Dioscorea Bulbifera Bulbis on Cyclophosphamide Induced Testicular Damage in Adult Male Wistar Rats

Cyclophosphamide is a synthetized drug and alkylating agent used for treating cancer. In this research, cyclophosphamide's effect on the testicles of adult male Wistar rats will be assessed, as well as the impact of alkaloid extract on the induced damage. Twenty-four healthy male wistar rats weighing between 132g to 168g of the same species of rattus Norvegicus were used. Rats were randomly assigned to 4 groups, A through D (n=7 in each). Group A was given distilled water. Group B were administered with Cyclophosphamide 150mg/ kg intra-peritoneal route for seven days. Group C was administered with Cyclophosphamide 150 mg/kg and 50 mg / kg of Alkaloid concomitantly for a week and Group D was administered Alkaloid for 7 days and then Cyclophosphamide of 150 mg /kg for 7 days. There was a significant difference in weight change, according to one way ANOVA result (F = 175.9; p &lt; 0.001), testosterone activity (F = 7.019; p = 0.0125), follicle stimulating hormone activity (F = 13.27; p = 0.0018), sperm motility (F = 11.95; p = 0.0025) in group B (cyclophosphamide only) as compared to control and across all experimental groups. Cyclophosphamide administration was observed to have a negative effect on the testicular histology and immunohistochemical results. The administration of alkaloids both concomitantly and as a pre-treatment helped to counteract the effect of the drug. In conclusion, the administration of cyclophosphamide should be strictly monitored and given in low doses alongside alkaloid to prevent toxic effect.

Comparative study of the reproductive toxicity and modulation of enzyme activities by crude oil-contaminated soil before and after bioremediation

Contamination of soil with crude oil is a serious ecological problem with potential adverse public health effects. This study assessed the germ cell toxicity of simulated leachates from crude oil-contaminated soil before and after bioremediation using the murine sperm abnormality assay, sperm count, and testes histopathology. The levels of Total Testosterone (TT), Follicle Stimulating Hormone (FSH), and Luteinizing Hormone (LH); and activities of catalase (CAT), alkaline phosphatase (ALP), superoxide dismutase (SOD), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) were determined. The physicochemical, Total Petroleum Hydrocarbon (TPH), and heavy metal analyses of the leachates were also carried out. Male mice were exposed to 50, 25, 10, 5, and 1% (v/v; leachate:distilled water) of the leachate samples for five consecutive days, and were sacrificed after 35 days. The result showed a statistically significant (p < 0.05), concentration-dependent increase in abnormal sperm cells in exposed mice, with aberrations such as folded sperm, amorphous head, wrong tail attachment, distal droplet, no hook, and looped tail. Data further showed a concentration-dependent significant reduction in mean sperm count in the exposed mice. Alterations of seminiferous tubules with different lesions and activities of ALT, AST, ALP, FSH, LH, and TT were also recorded. The high level of selected heavy metals (As, Cr, Cd, Cu, and Pb) and TPH was believed to contribute to the observed reproductive toxicity and modulated enzyme activities in the treated mice. It is therefore concluded that the microbial remediation of the crude oil contaminated soil produced a reduction in the levels of heavy metals and TPH in the soil, reduced reproductive toxicity, and modulation of enzyme activities. However, the induced reproductive toxicity by the bioremediated soil is still significant, hence, further work could be done to employ a consortium of bacteria and extend the period of the bioremediation process to ensure complete removal of the contaminants.

&lt;p&gt;&lt;strong&gt;Role of vitamin C on hormonal and pathological changes in &lt;em&gt;Heteropneustes fossilis&lt;/em&gt; (Bloch) due to exposure to sodium fluoride&lt;/strong&gt;&lt;/p&gt;

Fluoride is an essential trace element for humans and animals. Here, we explored the effects of sodium fluoride (NaF) on morphological, hormonal and histological changes in ovary of Asian stinging catfish Heteropneustes fossilis (Bloch) after exposure to 1/10th mg and 1/5th mg F/L of LC50 of fluoride in water for 45 and 90 days. Activities of follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T) and estradiol (E) were measured using an enzyme-linked immunosorbent assay. Histological evaluation of the ovaries was conducted after staining with haematoxylin-eosin (HE). Reproductive hormones were significantly decreased after NaF-treated groups. Ovary was found to be adversely effected by exposure to NaF. Maturation of follicles was inhibited by exposure after observation of GSI and histology in ovary. These results suggest that female reproductive system is adversely affected by NaF. NaF may thus significantly reduce the fertility of female fish. This study encompasses alteration in hormonal assay to evaluate the role of vitamins C in the recovery of fluoride toxicity in fish. Vitamin C is a water-soluble ROS scavenger with high potency.

Functions of Lysosomes in Mammalian Female Reproductive System

The lysosome is the most acidic membrane-bound intracellular organelle. Lysosomal acidity is primarily maintained by vacuolar H+-ATPase (V-ATPase) and counter ion channels. There are >60 hydrolytic enzymes in the lysosome for its fundamental digestive role. Lysosomes also play important roles in endocytosis, exocytosis, autophagy, and cell death. Studies that have implicated roles of lysosomes in the female reproductive system are reviewed here. In the ovary, lysosomes are implicated in the preparation of free cholesterol for steroidogenesis and degradation of regulators of steroidogenesis, regulation of follicular atresia, follicle rupture during ovulation, luteal cell survival, and luteal regression. In the oviduct, lysosomes are involved in endocytosis of both serum and oviductal luminal components. In the uterus during the menstrual/estrous cycle, lysosomes are associated with endometrial secretion, apoptosis, and menstruation. In the uterus during early pregnancy, lysosomes are involved in the temporal and directional changes of endocytosis, uterine epithelial acidification upon embryo implantation initiation, and embryo-maternal mutual communications via extracellular vesicles. In the placenta, lysosomes are implicated in nutrient transport and placental separation from the uterus for parturition. In the mammary gland, lysosomes are important for mammary gland development and involution. These findings suggest/demonstrate functions of lysosomes in multiple processes of female reproduction, from ovulation to ovarian steroidogenesis for pregnancy maintenance, and from essential in utero nurturing of developing embryos/fetuses via embryo/fetal-maternal communications, to optional postpartum nurturing of newborns via lactation. Future studies using genetically or modified animal models and pharmacological approaches will provide novel insights into the functions and mechanisms of lysosomes in the mammalian female reproductive system.

A case report supporting the concept that a role for the anti-Müllerian hormone (AMH) in normal folliculogenesis is to diminish the biological activity of follicle stimulating hormone (FSH)

Purpose: To describe a case where a woman was ovulating with regular menses despite very low sera follicle stimulating hormone (FSH) and luteinizing hormone (LH) levels. Case Report: Serum anti-Müllerian (AMH) was obtained because the woman was to be an egg donor for her identical twin sister. This 28-year-old woman responded fairly well to controlled ovarian hyperstimulation producing ten metaphase II eggs and eight fertilized and were cryopreserved on day 3. Her serum AMH level was subnormal at 0.6 ng/mL. Conclusions: This case suggests that FSH may be more biologically active in the presence of low AMH.

CYP19A1 Gene Expression in Patients with Polycystic Ovarian Syndrome.

Context:Polycystic ovarian syndrome (PCOS) is a common endocrine system disorder among the women of reproductive age, yet the etiology of PCOS remains unclear. Infertility in females with PCOS can be caused by anovulation, high luteinizing hormone levels, and hyperandrogenism.Aims:This research analyzed the role of the aromatase gene (CYP19A1) in PCOS pathogenesis.Settings and Design:This study used an observational, cross-sectional design.Subjects and Methods:A total of 110 research participants (55 PCOS patients and 55 non-PCOS patients) were included in the study.Statistical Analysis Used:A real-time quantitative polymerase chain reaction was used to analyze the mRNA expression for aromatase in granulosa cells.Results:The relative expression of aromatase mRNA is lower in women with PCOS compared to those without PCOS (P < 0.05). Relative expression of CYP19A1 (aromatase) mRNA in PCOS group was 0.38 ± 0.25, whereas in non-PCOS group was 1.00 ± 0.00. The decline in aromatase activity contributes to an increase in testosterone level. This condition has a role in hyperandrogenism which is a typical characteristic of PCOS women. Granulosa cells in polycystic ovary undergo disturbance in the development and cannot respond to follicle-stimulating hormone (FSH) stimulation. Lack of stimulation of FSH causes induction inadequacy to aromatase enzyme activity in the aromatization process. The decline in FSH activity is caused by various factors that are associated with typical characteristics of PCOS.Conclusions:There is a decrease in the relative expression rate of granulosa cells’ aromatase mRNA in women with PCOS compared to the non-PCOS.

Matrix metallopeptidases regulate granulosa cell remodeling through the hormone signaling pathway.

Objective:Granulosa cells (GCs) play a very important role in reproductive physiology due to their effect on developmental and functional changes. However, there are differing views regarding the mechanism by which hormones stimulate GCs. Therefore, our study aims to determine whether GCs, in the absence of initial stimulation (17β-estradiol), select specific types of MMPs that reconstitute cells by stimulation of major hormones [follicle-stimulating hormone (FSH) or/and luteinizing hormone (LH)].Materials and methods:Early GCs were extracted from immature follicles of the porcine ovary to analyze the MMPs levels. Using early GCs in pigs, the cell development rate was evaluated by adding 17β-estradiol, FSH, LH, or FSH + LH, respectively, to the DMEM containing 10% FBS. Real-time PCR, zymography, enzyme-linked immunosorbent assay, western blot, and immunofluorescence analysis were also performed to determine the MMPs activation in the GCs.Results:Our results confirm that FSH or LH stimulation regulates cell development and intracellular MMPs. In particular, FSH activity kept the MMP-2 and MMP-9 expressions constant in GCs. Conversely, LH activity initially led to rapid increases in the MMP-9 expression, which 96 h later was similar to the MMP-2 expression. Simultaneous utilization of FSH + LH maintained a steady MMP-9 expression and the development of GCs increased. Additionally, when FSH and LH were processed simultaneously, the number of cells increased without changes in cell size, while the cell size changed when LH alone was used.Conclusion:Therefore, the results of this study confirm that even without the initial stimulation of GCs, physiological changes occur according to hormonal changes in the environment, and there is variability in the expression of MMPs.

Anti-Mullerian Hormone Gene Expression and Serum Protein Concentration in Females Suffered from Diabetes Mellitus: An Animal Study

This study aimed to evaluate the effect of diabetes on fertility on diabetic women especially. Sixty adult female patients aged (18 – 40) years are divided equally into three groups; G1; control healthy group, G2; type one diabetic women, G3; non fertile women. Blood samples of all individuals were analyzed for measurement of glucose, estradiol, progestrone, anti-mullerian hormone (AMH), Follicle stimulating hormone (FSH), Luteinizing hormone (LH), superoxide dismutase (SOD) and nitric oxide (NO), serum insulin and glycated hemoglobin (HbA1c) showed a significant increase in diabetic fertile group. Also plasma level of FSH, LH and SOD activity were significantly lowered in diabetic infertile group than healthy group and diabetic fertile group. NO was higher in diabetic fertile group. In AMH mRNA expression in ovarian tissues, 40 adult female Sprague-Dawley rats were divided equally into four groups; G1; normal rats, G2; diabetic 30 days group rat, G3; diabetic 45 days group rat, G4; diabetic 60 days group rat. the expression of AMH in ovaries was significantly increased in diabetic rats at day 30 compared to healthy rats and significantly decreased at days 45 and 60 compared to healthy rats. In conclusion, diabetes mellitus can affect the health status and fertility markers in females that was also observed in expression of AMH in ovarian tissues in rats

Glycosylation Pattern and in vitro Bioactivity of Reference Follitropin alfa and Biosimilars.

Recombinant follicle-stimulating hormone (FSH) (follitropin alfa) and biosimilar preparations are available for clinical use. They have specific FSH activity and a unique glycosylation profile dependent on source cells. The aim of the study is to compare the originator (reference) follitropin alfa (Gonal-f®)- with biosimilar preparations (Bemfola® and Ovaleap®)-induced cellular responses in vitro. Gonadotropin N-glycosylation profiles were analyzed by ELISA lectin assay, revealing preparation specific-patterns of glycan species (Kruskal-Wallis test; p < 0.05, n = 6) and by glycotope mapping. Increasing concentrations of Gonal-f® or biosimilar (1 × 10−3-1 × 103 ng/ml) were used for treating human primary granulosa lutein cells (hGLC) and FSH receptor (FSHR)-transfected HEK293 cells in vitro. Intracellular cAMP production, Ca2+ increase and β-arrestin 2 recruitment were evaluated by BRET, CREB, and ERK1/2 phosphorylation by Western blotting. 12-h gene expression, and 8- and 24-h progesterone and estradiol synthesis were measured by real-time PCR and immunoassay, respectively. We found preparation-specific glycosylation patterns by lectin assay (Kruskal-Wallis test; p < 0.001; n = 6), and similar cAMP production and β-arrestin 2 recruitment in FSHR-transfected HEK293 cells (cAMP EC50 range = 12 ± 0.9–24 ± 1.7 ng/ml; β-arrestin 2 EC50 range = 140 ± 14.1–313 ± 18.7 ng/ml; Kruskal-Wallis test; p ≥ 0.05; n = 4). Kinetics analysis revealed that intracellular Ca2+ increased upon cell treatment by 4 μg/ml Gonal-f®, while equal concentrations of biosimilars failed to induced a response (Kruskal-Wallis test; p < 0.05; n = 3). All preparations induced both 8 and 24 h-progesterone and estradiol synthesis in hGLC, while no different EC50s were demonstrated (Kruskal-Wallis test; p > 0.05; n = 5). Apart from preparation-specific intracellular Ca2+ increases achieved at supra-physiological hormone doses, all compounds induced similar intracellular responses and steroidogenesis, reflecting similar bioactivity, and overall structural homogeneity.

Quercetin directly inhibits basal ovarian cell functions and their response to the stimulatory action of FSH

Plants, fruits, and vegetables containing the bioflavonoid quercetin are widely used in food, beverages, and medicines; however, the effects of quercetin on reproductive processes and the possible mechanisms of quercetin action require extensive investigation. The aim of our study was to examine the direct effects of quercetin on basic ovarian cell functions and their response to follicle-stimulating hormone (FSH) and insulin-like growth factor I (IGF-I), known hormonal stimulators of reproduction. We analyzed the effects of quercetin alone (0, 1, 10, and 100 ng/ml) on cultured porcine ovarian granulosa cells or isolated ovarian follicles; or of quercetin (10 ng/ml) in combination with FSH (0, 0.01, 0.1, or 1 IU/ml) or IGF-I (0, 1, 10, or 100 ng/ml) on cultured porcine granulosa cells. The expression of proliferative (PCNA, cyclin B1) and apoptotic (BAX) markers, as well as markers for release of progesterone (P4), testosterone (T), and leptin (L), were measured by quantitative immunocytochemistry, Western immunoblotting, RT-qPCR, and EIA/RIA. Addition of quercetin reduced the accumulation of PCNA and cyclin B1, as well as their transcript levels, promoted the accumulation of BAX, decreased the release of P4 and L, and increased the release of T in cultured granulosa cells. In ovarian follicles, quercetin reduced the levels of both P4 and T. Exposure to FSH stimulated PCNA and decreased BAX accumulation, and increased the release of P4, T, and L. Quercetin inhibited and even reversed the effects of FSH. Like FSH, IGF-I also promoted granulosa cell proliferation and suppressed apoptosis. Quercetin did not modify IGF-I effects. These data suggest that the plant molecule quercetin can directly down-regulate basal ovarian cell functions (proliferation, apoptosis, and release of ovarian steroid and peptide hormones) and their response to the stimulatory activity of the upstream hormonal stimulator FSH.

Evaluation of the serum sex hormones levels and alkaline phosphatase activity in rats’ testis after administering of berberine in experimental varicocele

Current study was aimed to investigate the protective effect of berberine (BB) on the serum gonadotropin-releasing hormone (GnRH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), inhibin B (INHB), testosterone (T) and alkaline phosphatase (Alk-p) activity in the testis of experimental varicocele-induced animals. For the current objective, 30 mature-male Wistar rats were randomly divided into control (n = 6 rats), control-sham (n = 6 rats) and experimental groups (n = 18 rats). The animals in the experimental groups were undergone experimental varicocele and simple laparotomy was conducted in control-sham group. 60 days after varicocele (VCL) induction the experimental group subdivided into: non-treated VCL-induced and 50 mg/kg and 100 mg/kg BB-treated groups (intra-peritoneally). Following 60 days, the animals were euthanized and serum levels of testosterone and testicular activity of alkaline phosphatase were measured. Non-treated VCL-induced animals indicated a significant (P < 0.05) reduction in serum levels of T and INHB and a remarkable (P < 0.05) increase in GnRH, FSH, LH and Alk-p activity compared to control and control-sham groups. Insignificant changes were found between control and control-sham groups. Meanwhile, each BB administered group showed a remarkable (P < 0.05) increase in serum levels of T and INHB and a significant (P < 0.05) decrease in GnRH, FSH, LH and alkaline phosphatase activity in testis tissue. According to the current findings, BB by increasing serum levels of testosterone and INHB increases the testicular endocrine capacity and protects Leydig cell against inflammatory and oxidant injury of varicocele. In addition, BB by inhibiting GnRH, FSH, LH and alkaline phosphatase activity, regulate the levels of serum sex hormones in experimental varicocele and reduces varicocele-induced inflammatory reactions.

Effect of Polycyclic Aromatic Hydrocarbons Exposure on Sperm DNA in Idiopathic Male Infertility.

Background.Biological mechanisms contribute to the relationship between polycyclic aromatic hydrocarbon (PAH) exposure and infertility in males by altering semen quality.Objectives.The aim of the present study was to evaluate the impact of PAHs on male infertility using the sperm chromatin dispersion test (Halo sperm assay).Methods.Sixty-six (66) infertile males under 45 years of age were examined for the determination of urinary metabolite and oxidative stress by measuring lipid peroxidation and antioxidant activity of glutathione and glutathione-s-transferase, as well as hormonal activity of follicle stimulating hormone (FSH), testosterone and prolactin and semen quality.Results.There was an increased level of urinary metabolite of 1-hydroxy pyrene, 1-hydroxy naphthalene and 2-hydroxy naphthalene in the urine of the infertile group. In addition, elevated concentrations of malondialdehyde coincided with a decreased level of antioxidants, leading to oxidative stress in the infertile group. Semen samples showed 30% sperm deoxyribonucleic acid (DNA) fragmentation.Conclusions.The data provide strong evidence of a statistical threshold for semen samples containing 30% sperm DNA fragmentation resulting in a reduced level of pregnancy success.Participant consent.ObtainedEthics Approval.Study approval was given by the ethics committee of Alexandria University (United States Department of Health and Human Services, institutional review board registration (IRB), IORG0008812 Medical Research Institute, expires 4/8/2019, OMB No: 0990-0279).Competing Interests.The authors declare no competing financial interests.

Effects of recombinant vertebrate ancient long opsin on reproduction in goldfish, Carassius auratus: profiling green-wavelength light.

This study was conducted to identify the possible effect of recombinant vertebrate ancient long (VAL) opsin as a non-visual "photoreceptor" in the deep brain of goldfish, Carassius auratus. In addition, we investigated the effects of green-wavelength light on the predictable reproductive function of VAL-opsin as a green-sensitive pigment in the deep brain. To determine this, we quantified changes in gonadotropin hormone (GTH) [GTHα, follicle stimulating hormone (FSH) and luteinizing hormone (LH)] and estrogen receptor (ER; ERα and ERβ) mRNA expression levels associated with goldfish reproduction as well as changes in plasma FSH, LH, and 17β-estradiol (E2) activities after injection of recombinant VAL-opsin protein in two concentrations (0.1 or 0.5μg/g body mass) for 4weeks (injection once weekly) and examined the possible impact of green-wavelength light (500, 520, and 540nm) on the function of VAL-opsin. As a result, all parameters associated with reproduction significantly increased with time and light-emitting diode (LED) exposure. Based on these results, we suggested that VAL-opsin in the deep brain is involved in goldfish maturation, and it is possible that green-wavelength light improves the ability of VAL-opsin to promote maturation by increasing VAL-opsin expression.

The switch from cAMP-independent to cAMP-dependent arrest of meiotic prophase is associated with coordinated GPR3 and CDK1 expression in mouse oocytes

Mammalian oocytes are arrested in meiotic prophase from around the time of birth until just before ovulation. Following an extended period of growth, they are stimulated to mature to the metaphase II stage by a preovulatory luteinizing hormone (LH) surge that occurs with each reproductive cycle. Small, growing oocytes are not competent to mature into fertilizable eggs because they do not possess adequate amounts of cell cycle regulatory proteins, particularly cyclin-dependent kinase 1 (CDK1). As oocytes grow, they synthesize CDK1 and acquire the ability to mature. After oocytes achieve meiotic competence, meiotic arrest at the prophase stage is dependent on high levels of cAMP that are generated in the oocyte under the control of the constitutively active Gs-coupled receptor, GPR3. In this study, we examined the switch between GPR3-independent and GPR3-dependent meiotic arrest. We found that the ability of oocytes to mature, as well as oocyte CDK1 levels, were dependent on follicle size, but CDK1 expression in oocytes from preantral follicles was not acutely altered by the activity of follicle stimulating hormone (FSH). Gpr3 was expressed and active in incompetent oocytes within early stage follicles, well before cAMP is required to maintain meiotic arrest. Oocytes from Gpr3-/- mice were less competent to mature than oocytes from Gpr3+/+ mice, as assessed by the time course of germinal vesicle breakdown. Correspondingly, Gpr3-/- oocytes contained significantly lower CDK1 levels than their Gpr3+/+ counterparts that were at the same stage of follicle development. These results demonstrate that GPR3 potentiates meiotic competence, most likely by raising cAMP.

Oxidative stress status in congenital hypogonadism: an appraisal

Patients with hypogonadism are at increased risk of cardiac and metabolic diseases. However, the pathogenesis of increased cardiometabolic risk in patients with hypogonadism is not clear. Oxidative stress plays an important role in the pathogenesis of cardiometabolic diseases. This study aimed to investigate possible differences in oxidative stress conditions between patients with hypogonadism and healthy controls. In this study, 38 male patients with congenital hypogonadotropic hypogonadism (CHH) (mean age: 21.7 ± 1.6 years) and 44 healthy male controls (mean age: 22.3 ± 1.4 years) with almost equal body mass index were enrolled. The demographic parameters, follicle-stimulating hormone (FSH), luteinizing hormone (LH), total and free testosterone, homeostatic model assessment of insulin resistance (HOMA-IR) and oxidative stress parameters, such as superoxide dismutase, catalase (CAT), glutathione peroxidase (GPx) and malondialdehyde (MDA), were compared between both groups. Compared to the healthy controls, triglycerides (p = .02), insulin levels, HOMA-IR values, CAT activities and MDA levels (p < .001 for all) were significantly higher and HDL cholesterol (p = .04), total and free testosterone, FSH, LH levels and GPx activity were significantly lower (p < .001 for all) in patients with CHH. There were significant correlations between total testosterone levels and CAT activity (r = −.33 p = .01), GPx activity (r = .36 p = .007) and MDA (r = −.47 p < .001) levels. The results of this study showed that young and treatment-naïve patients with congenital hypogonadism had an increased status of oxidative stress.

Determination of biological activity of gonadotropins hCG and FSH by F\xf6rster resonance energy transfer based biosensors

Determination of biological activity of gonadotropin hormones is essential in reproductive medicine and pharmaceutical manufacturing of the hormonal preparations. The aim of the study was to adopt a G-protein coupled receptor (GPCR)-mediated signal transduction pathway based assay for quantification of biological activity of gonadotropins. We focussed on studying human chorionic gonadotropin (hCG) and follicle-stimulating hormone (FSH), as these hormones are widely used in clinical practice. Receptor-specific changes in cellular cyclic adenosine monophosphate (cAMP, second messenger in GPCR signalling) were monitored by a Förster resonance energy transfer (FRET) biosensor protein TEpacVV in living cells upon activation of the relevant gonadotropin receptor. The BacMam gene delivery system was used for biosensor protein expression in target cells. In the developed assay only biologically active hormones initiated GPCR-mediated cellular signalling. High assay sensitivities were achieved for detection of hCG (limit of detection, LOD: 5 pM) and FSH (LOD: 100 pM). Even the small-scale conformational changes caused by thermal inactivation and reducing the biological activity of the hormones were registered. In conclusion, the proposed assay is suitable for quantification of biological activity of gonadotropins and is a good alternative to antibody- and animal-testing-based assays used in pharmaceutical industry and clinical research.

The Observation of FSH’s Cellular Internalization

Follicle stimulating hormone (FSH) is a kind of glycoprotein gonadotropin, and plays an important role in the diagnosis and treatment of infertility. Follicle stimulating hormone receptor (FSHR) is a kind of G protein coupled receptor (GPCR), found in the ovary and testes, and its activation is required for the hormonal operation during the breeding period. In this study, an experimental model of FSHR mediated FSH into cell membrane, which exhibited a phenomenon of fluorescent localized on cell surfaces internalized into cell interior, was established to verify biological activity of FSH.