Role Of Gonadotropins Research Articles

Medical therapy for male infertility.

To provide up-to-date evidence and clinical guidance on the role of medical therapy in the context of hormonal imbalances affecting human spermatogenesis. Compelling evidence has accumulated over the years regarding the role of gonadotropins, selective estrogen modulators, and aromatase inhibitors to either improve or restore spermatogenesis in men with hormonal abnormalities (e.g. hypogonadotropic/hypergonadotropic hypogonadism, hyperprolactinemia) or supraphysiologic levels (e.g. exogenous testosterone/anabolic steroid use). Despite the increasing number of studies being performed, most of the available evidence relies on small nonrandomized studies, mainly in men with hypergonadotropic hypogonadism or with history of exogenous testosterone/anabolic steroid use. As such, the efficacy of medical therapy is highly variable emphasizing the necessity of randomized clinical trials and individualized approaches. This narrative review provides clinical guidance on medical therapies for male factor infertility based on the most up-to-date evidence, focusing on treatments for hormonal abnormalities (either hypogonadotropic or hypergonadotropic hypogonadism and hyperprolactinemia) and supraphysiologic levels (and exogenous testosterone/anabolic steroid use) to improve spermatogenesis.

Ovarian mRNA Expression and Regulation of Matrix Metalloproteinase 16 in the Domestic Hen

In mammals, membrane-bound matrix metalloproteinases (MT-MMPs) are thought to play an important role in ovarian remodeling. However, the role and regulation of these proteases in the ovary of birds remain largely unknown. One of MT-MMPs, i. e., MMP-16, has been found in the hen ovary; therefore, this study was undertaken to examine whether the transcript level of MMP-16 changes during follicle development and whether gonadotropins and estrogen are involved in the regulation of this enzyme expression. The relative expression of MMP-16 mRNA in the ovarian follicles (white, yellowish, small yellow, and the granulosa and theca layers of three of the largest yellow preovulatory [F3-F1]) was examined 22 h and 3 h before F1 follicle ovulation as well as following equine chorionic gonadotropin (eCG) or tamoxifen (estrogen receptor modulator, TMX) treatments by quantitative real-time polymerase chain reaction (qRT-PCR). MMP-16 transcripts were detected in all examined ovarian tissues of control and treated hens. The relative expression of MMP-16 depended on follicular size/maturation and the layer of the follicular wall. A relatively higher expression of MMP-16 mRNA in the granulosa layer at 3 h compared to 22 h before ovulation of F1 was found. The injections of eCG decreased transcript abundance of MMP-16 in white and small yellow follicles, as well as in the theca layer of F3-F2 and the granulosa layer of the F1 follicle. In turn, TMX caused an increase in mRNA expression of MMP-16 in the theca layer of the largest preovulatory follicles and a decrease in the granulosa layer of the F1 follicle. Our results provide the first mRNA expression analysis of MMP-16 in the hen ovary under different physiological states. In addition, results indicate a possible role of gonadotropins and estrogen in regulating the transcription of MMP-16 in the chicken ovary.

Estropause, Sex Hormones and Metal Homeostasis in the Mouse Brain.

Alterations in brain metal ion homeostasis have been reported with aging and are implicated in the pathogenesis of neurodegenerative diseases. To assess whether age-related changes in hypothalamic-pituitary-gonadal (HPG) hormones might be involved in modulating brain metal ion homeostasis, we treated 7.5-month intact, sham-ovariecomized and ovariectomized C57B6SJL mice with vehicle or leuprolide acetate (for 9-months) to differentiate between whether sex steroids or gonadotropins might modulate brain metal ion concentrations. Unlike other aging mammals, there was no increase in plasma luteinizing hormone (LH) and follicle-stimulating hormone (FSH) concentrations following estropause in mice, suggesting there was sufficient residual production by the follicle depleted ovary, of sex steroids like estrogens and protein hormones like the inhibins, in order to suppress pituitary LH/FSH production. Castration on the other hand induced significant increases in circulating LH and FSH. Modulation of plasma sex steroid and gonadotropin levels did not significantly alter the concentrations of brain metals tested (Fe, Zn, Cu, Mn, Co, Ni, Al, Li), although there was a tendency for a decrease in all brain metals following ovariectomy (low estrogens and progesterone, high gonadotropins), a response that was reversed with leuprolide acetate treatment (low sex steroids, low gonadotropins). Brain Cu concentration was the only metal correlated with plasma LH (−0.37, n = 30, p < 0.05) and FSH (−0.42, n = 29, p < 0.01). This study demonstrates that sex hormones do not markedly alter brain metal ion homeostasis, unlike previously reported studies of circulating metal ion homeostasis. The role of gonadotropins in regulating metal ion homeostasis does however warrant further study.

The role of gonadotrophins in gonocyte transformation during minipuberty.

Postnatal surge of gonadotrophins, Luteinizing hormone (LH) and Follicle-Stimulating hormone (FSH) known as minipuberty, is critical for gonocyte maturation into spermatogonial stem cells (SSC) in the testis. Gonadotrophins are essential for optimum fertility in men, but very little is known how they regulate germ cells during minipuberty. This study examined whether gonadotrophins play a role on gonocyte transformation in vivo. Testes from hypogonadal (hpg) mice and their wild type (WT) littermates (n = 6/group) were weighed, and processed in paraffin at postnatal days (D) 0, 3, 6 and 9. Mouse VASA homologue (germ cell marker), anti-Müllerian hormone (Sertoli cell marker) antibodies and DAPI (nuclei marker) were used for immunofluorescence followed by confocal imaging. Germ cells on or off basement membrane (BM) and Sertoli cells/tubule were counted using Image J and analyzed with GraphPad. Comparing to WT littermates, there were significantly fewer germ cells on BM/tubule (p < 0.05) in D9 hpg mice, whereas there was no significant difference for germ cells off BM/tubule and Sertoli cells/tubule between littermates. However, testicular weight was significantly reduced in D3-D9 hpg mice comparing to WT littermates. Gonadotrophin deficiency reduced D9 germ cells on BM indicating impaired gonocyte transformation into SSC. This suggests that gonadotrophins may mediate gonocyte transformation during minipuberty.

Induction of oocyte development in previtellogenic eel, Anguilla australis

The role of gonadotropins during early ovarian development in fish remains little understood. Concentrations of gonadotropins were therefore experimentally elevated in vivo by administration of recombinant follicle-stimulating hormone (rec-Fsh) or human chorionic gonadotropin (hCG) and the effects on ovarian morphology, sex steroid levels and mRNA levels of genes expressed in pituitary and ovary examined. Hormones were injected thrice at weekly intervals in different doses (20, 100 or 500 µg/kg BW for rec-Fsh and 20, 100 or 500 IU/kg BW for hCG). All treatments, especially at the highest doses of either rec-Fsh or hCG, induced ovarian development, reflected in increased oocyte size and lipid uptake. Both gonadotropins up-regulated follicle-stimulating hormone receptor (fshr) mRNA levels and plasma levels of estradiol-17β (E2). Exogenous gonadotropins largely decreased the expression of follicle-stimulating hormone β-subunit (fshb) and had little effect on those of luteinizing hormone β-subunit (lhb) in the pituitary. It is proposed that the effects of hCG on ovarian development in previtellogenic eels could be indirect as a significant increase in plasma levels of 11-ketotestosterone (11-KT) was found in eels treated with hCG. Using rec-Fsh and hCG has potential for inducing puberty in eels in captivity, and indeed, in teleost fish at large.

Gonadotropins promote human ovarian cancer cell migration and invasion via a cyclooxygenase 2-dependent pathway.

It is generally accepted that ovarian cancer is associated with local elevation of gonadotropins (FSH and LH), with repeated ovulation and accompanying expression of inducible cyclooxygenase2 (COX2). However, the roles of gonadotropins and the concomitant elevation of COX2 in the development of ovarian cancer have not been fully characterized. Herein, we report that excessive FSH/LH exposure did not induce proliferation in ovarian cancer cell lines but significantly promoted cell migration and invasion. Moreover, FSH/LH treatment rapidly upregulated COX2 expression within 24h, whereas COX1 expression remained unchanged. Further results showed that enhancement of epithelial-mesenchymal transition (EMT) and upregulation of matrix metalloproteinase (MMP)2 and MMP9 contributed to the stimulatory effect of gonadotropins on cell migration and invasion; these effects were sufficiently blocked by a selective COX2 inhibitor. In conclusion, the present study suggests that gonadotropin-induced migration and invasion in ovarian cancer may be caused by EMT and MMP upregulation via a COX2-dependent pathway.

Changes in plasma steroid levels during oocyte development in Indian shad, Tenualosa ilisha (Hamilton, 1822): Role of gonadotropins on in vitro steroid production and development of oocyte maturational competence

Circanual variations in plasma testosterone (T), 17-estradiol (E2), and 17,20-dihydroxy-4-pregnen-3-one (17,20-P) levels and ovarian steroid synthetic potential of Tenualosa ilisha of river Hooghly, West Bengal, India were examined. This fish exhibited bi-annual spawning; one during April–May and another during August–September. Coinciding with the GSI values, present study recorded a decline in plasma T and E2 levels from October, reaching their lowest values in January followed by a rapid rise in March when the ovary contained mostly vitellogenic follicles and remained high up to April (postvitellogenic stage). Plasma 17,20β-P level was detected in March and reached peak value in April during oocyte maturation. After spawning, all the steroid levels declined to reach lowest values in June. From June onwards, T and E2 levels again increased for the next cycle and peaked at the end of vitellogenesis. Plasma 17,20β-P was reappeared in August and reached maximum in September during oocyte maturation and spawning. Of the two gonadotropins tested, in vitro production of both T and E2 by the vitellogenic and postvitellogenic follicles was regulated by FSH and LH respectively. Production of 17,20-P by the post-vitellogenic follicles was regulated by LH only. Acquisition of in vitro oocyte maturational competence (OMC) was developed by the addition of HCG in culture medium. Treatment of a 3β-HSD inhibitor blocked LH-induced steroid production, but not development of OMC. Both Cycloheximide and actinomycin D inhibited LH-induced development of OMC, indicating the requirement of de novo protein synthesis for this process.

Expression of Estrogen Receptor α 36 (ESR36) in the Hamster Ovary throughout the Estrous Cycle: Effects of Gonadotropins

Estradiol-17β (E) plays an important role in ovarian follicular development. Evidence indicates that some of the effect of E is mediated by the transmembrane estrogen receptor. In this study, we examined the spatio-temporal expression of recently discovered ERα36 (ESR36), a splice variant of Esr1 and a receptor for non-genomic E signaling, in the hamster ovary during the estrous cycle and the role of gonadotropins and ovarian steroid hormones in ESR36 expression. ESR36 expression was high on estrus (D1∶0900 h) and declined precipitously by proestrus (D4∶0900 h) and remained low up to D4∶1600 h. Immunofluorescence findings corroborated immunoblot findings and revealed that ESR36 was expressed only in the cell membrane of both follicular and non-follicular cells, except the oocytes. Ovarian ESR36 was capable of binding to the E-affinity matrix, and have different molecular weight than that of the ESR1 or GPER. Hypophysectomy (Hx) resulted in a marked decline in ESR36 protein levels. FSH and LH, alone or combined, markedly upregulated ESR36 protein in Hx hamsters to the levels observed in D1 hamsters, but neither E nor P had any effect. Inhibition of the gonadotropin surge by phenobarbital treatment on D4∶1100 h attenuated ESR36 expression in D1∶0900 h ovaries, but the decline was restored by either FSH or LH replacement on D4 afternoon. This is the first report to show that ESR36, which is distinct from ESR1 or GPER is expressed in the plasma membrane of ovarian follicular and non-follicular cells, binds to E and its expression is regulated directly by the gonadotropins. In light of our previous findings, the results suggest that ovarian cells contain at least two distinct membrane estrogen receptors, such as GPER and ESR36, and strongly suggest for a non-genomic action of E regulating ovarian follicular functions.

Testosterone modulation of anxiety in gonadally-suppressed male rhesus monkeys: A role for gonadotropins?

Testosterone (T) has repeatedly been shown to have anxiolytic properties in rodents, but findings in primates are more mixed. To examine the effects of exogenous T on anxiety, we tested pharmacologically-castrated adult male rhesus monkeys in a modified version of the Human Intruder Paradigm, which measured defensive responses to an unfamiliar human staring directly at them for 2min. Monkeys were tested at 2week intervals during 4 experimental conditions lasting 4weeks each: at baseline, during treatment with the gonadotropin releasing hormone (GnRH) agonist leuprolide acetate (200μg/kg; Lupron phase), during treatment with Lupron+T enanthate (TE, 5mg/kg; TE phase) and during treatment with Lupron+oil vehicle (oil phase). We found that the number of anxious behaviors was lower during periods of low T (Lupron only and Lupron+oil phases) than during the Lupron+TE phase. No change in pacing or watching behavior was observed. Thus, in contrast to rodent data, we found no evidence for anxiolytic properties of T in male rhesus monkeys. Rather, T supplementation restored baseline levels of anxiety in Lupron-treated monkeys. These discrepant findings may be best explained by the low levels of gonadotropins achieved by the GnRH agonist. We suggest that Lupron-induced luteinizing hormone (LH) suppression reduced anxiety and that this effect was abolished by T administration. This interpretation is consistent with the view that T increases emotional reactivity to a potential threat and facilitates adaptive arousal response in face of immediate social challenge.

Gonadotropins Are Related to Lean Mass in Healthy Postmenopausal Women

Background. It has been suggested that the changes in hormone levels that occur with menopause and aging are related to loss of muscle mass. However, the role of gonadotropins in this process is unknown. Objectives. To analyze the relationship between follicle-stimulating hormone (FSH) and luteinizing hormone (LH) and lean mass in a cohort of healthy women. Methods. This is a cross-sectional study including 91 postmenopausal women (48–65 years old). Serum FSH, LH, and estradiol (E2) were measured. Lean mass was estimated by bioelectrical impedance analysis and expressed as appendicular skeletal muscle mass (AMM) index. Results. AMM index was negatively correlated with FSH levels (r = −0.33, p = 0.001) and LH (r = −0.293, p = 0.005), while there was no association with E2. When muscle mass expressed as AMM index was stratified into tertiles, there were significant differences between the lowest and the highest tertile for both FSH (82.4 ± 30, vs. 67.9 ± 25.8 mIU/mL, p = 0.04) and LH (40.9 ± 15.2 vs. 36.1 ± 12.3 mIU/mL, p = 0.03). In logistic regression analysis adjusted for age and E2 levels, high concentrations of FSH (OR = 1.03, 95% CI = 1.01–1.05, p = 0.008) and LH (OR = 1.06, 95% CI = 1.01–1.1, p = 0.01) were risk factors for having lower lean mass expressed as AMM index. Conclusions. Menopausal women with high levels of gonadotropins have lower levels of lean mass than those with lower gonadotropins levels.

Up-regulation of FSHR expression during gonadal sex determination in the frog Rana rugosa

In vertebrates, gonadal production of steroid hormones is regulated by follicle-stimulating hormone (FSH) and luteinizing hormone (LH) via their receptors designated FSHR and LHR, respectively. We have shown recently that steroid hormones are synthesized in the differentiating gonad of tadpoles during sex determination in the frog Rana rugosa. To elucidate the role of gonadotropins (GTHs) and their receptors in the production of gonadal steroid hormones during sex determination, we isolated the full-length FSHβ, LHβ, FSHR and LHR cDNAs from R. rugosa and determined gonadal expression of FSHR (FSH receptor) and LHR (LH receptor) as well as brain expression of FSHβ and LHβ during sex determination in this species. The molecular structures of these four glycoproteins are conserved among different classes of vertebrates. FSHβ expression was observed at similar levels in the whole brain (including the pituitary) of tadpoles, but it showed no sexual dimorphism during gonadal sex determination. By contrast, LHβ mRNA was undetectable in the whole brain of tadpoles. FSHβ-immunopositive cells were observed in the pituitary of female tadpoles with a differentiating gonad. Furthermore, FSHR expression was significantly higher in the gonad of female tadpoles during sex determination than in that of males, whereas LHR was expressed at similar levels in males and females. The results collectively suggest that FSHR, probably in conjunction with FSH, is involved in the steroid-hormone production during female-sex determination in R. rugosa.

The role of gonadotropins in the follicular phase

Folliculogenesis in humans is a lengthy process that involves several regulators. Pituitary gonadotropins play crucial roles in the late stages, particularly in the last 15 days of follicle maturation. During the intercycle rise of follicle-stimulating hormone (FSH), selection of the dominant follicle takes place. This is a complex process that also involves locally produced substances. At the same time, luteinizing hormone (LH) stimulates the synthesis of androgens, which serve as the substrate for the production of estrogens. During the second half of the follicular phase, the follicle becomes dependent on LH. Induction of multiple follicular development by exogenous FSH results in a marked suppression of endogenous LH. For normal follicle maturation, both an LH threshold and an LH ceiling have been considered. In the context of an in vitro fertilization program, application of protocols for ovarian stimulation that will prevent the marked suppression of endogenous LH secretion might provide a better approach to treatment optimization.

Survival, growth, and maturation of secondary follicles from prepubertal, young, and older adult rhesus monkeys during encapsulated three-dimensional culture: effects of gonadotropins and insulin

A three-dimensional culture system supports the development of primate preantral follicles to the antral stage with appreciable steroid production. This study assessed i) whether in vitro developmental competence of follicles is age dependent, ii) the role of gonadotropins and insulin in supporting folliculogenesis, and iii) anti-Müllerian hormone (AMH) and vascular endothelial growth factor (VEGF) production by growing follicles. Ovaries were obtained from prepubertal, young, and older adult rhesus macaques. Secondary follicles were encapsulated into alginate beads and cultured individually for 40 days in media containing 0.05 or 5 μg/ml insulin, with or without recombinant human (rh) FSH (500 mIU/ml). No follicles survived in the culture without rhFSH. In the presence of rhFSH, survival was lower for follicles from older animals, whereas growth, i.e. follicle diameter, was less by day 40 for follicles from prepubertal animals. The surviving follicles were categorized as no-grow (NG; ≤ 250 μm), slow-grow (SG; 250-500 μm), and fast-grow (FG; ≥ 500 μm) according to their diameters. SG follicles cultured with 5 μg/ml insulin produced more ovarian steroids than those cultured with 0.05 μg/ml insulin by week 5. SG and FG follicles produced more AMH and VEGF than the NG, and levels peaked at weeks 2 and 5 respectively. After 100 ng/ml rh chorionic gonadotropin treatment for 34 h, more healthy oocytes were retrieved from young adults whose follicles were cultured with 5 μg/ml insulin. This culture system offers an opportunity to characterize the endocrine and paracrine function of primate follicles that influence follicle growth and oocyte maturation.

Gonadotropins Induce Tumor Cell Migration and Invasion by Increasing Cyclooxygenases Expression and Prostaglandin E2Production in Human Ovarian Cancer Cells

Gonadotropins (FSH and LH) are detectable in ovarian tumor fluid, suggesting a possible role for gonadotropins in ovarian carcinogenesis and progression. However, the molecular mechanisms behind the role of gonadotropins in ovarian cancer development are unknown. Cyclooxygenase (COX) enzymes, COX-1 and COX-2, play crucial roles in the pathogenesis of human malignancies. The purpose of the current study was to determine whether the effect of gonadotropins on ovarian cancer invasion is mediated by a COX-dependent mechanism. Here, we reported that FSH/LH can promote prostaglandin E(2) (PGE(2)) production in ovarian cancer cells via COX-1 and -2 up-regulation at the protein and mRNA level. The phosphatidylinositol-3-kinase (PI3K)/AKT signaling pathway was required for gonadotropin-mediated up-regulation of COX-1 and COX-2. Moreover, treatment with COX-1- and COX-2-specific inhibitors abrogated the stimulatory effect of gonadotropins on motility and invasive activity. Western blot and gelatin zymography showed that COX-1 and COX-2 were critical for gonadotropin-induced expression of metastasis-related proteinases, matrix metalloproteinase (MMP)-2 and MMP-9. In addition, our results showed that PGE(2) induced an increase in cell invasiveness and the expression of MMP-2 and MMP-9 in ovarian cancer cells. These data show that COX-1 and COX-2 play essential roles in gonadotropin-induced migration and invasion.

Gonadotropins Regulate Rat Testicular Tight Junctions in Vivo

Sertoli cell tight junctions (TJs) are an essential component of the blood-testis barrier required for spermatogenesis; however, the role of gonadotropins in their maintenance is unknown. This study aimed to investigate the effect of gonadotropin suppression and short-term replacement on TJ function and TJ protein (occludin and claudin-11) expression and localization, in an adult rat model in vivo. Rats (n = 10/group) received the GnRH antagonist, acyline, for 7 wk to suppress gonadotropins. Three groups then received for 7 d: 1) human recombinant FSH, 2) human chorionic gonadotropin (hCG) and rat FSH antibody (to study testicular androgen stimulation alone), and 3) hCG alone (to study testicular androgen and pituitary FSH production). TJ proteins were assessed by real-time PCR, Western blot analysis, and immunohistochemistry, whereas TJ function was assessed with a biotin permeation tracer. Acyline treatment significantly reduced testis weights, serum androgens, LH and FSH, and adluminal germ cells (pachytene spermatocyte, round and elongating spermatids). In contrast to controls, acyline induced seminiferous tubule permeability to biotin, loss of tubule lumens, and loss of occludin, but redistribution of claudin-11, immunostaining. Short-term hormone replacement stimulated significant recoveries in adluminal germ cell numbers. In hCG +/- FSH antibody-treated rats, occludin and claudin-11 protein relocalized at the TJ, but such relocalization was minimal with FSH alone. Tubule lumens also reappeared, but most tubules remained permeable to biotin tracer, despite the presence of occludin. It is concluded that gonadotropins maintain Sertoli cell TJs in the adult rat via a mechanism that includes the localization of occludin and claudin-11 at functional TJs.

Expression profiles of LHβ, FSHβ and their gonadal receptor mRNAs during sexual differentiation of Xenopus laevis tadpoles

The gonadotropins, luteinising hormone (LH) and follicle stimulating hormone (FSH), are important hormones regulating reproductive biology in vertebrates, especially the processes of steroidogenesis and gamete maturation. Despite the role of gonadotropins during the reproductive cycle in amphibians is well established, much less is known about the functional maturation of the hypothalamus–pituitary–gonad axis during larval development. Therefore, the present study aimed to analyze the expression profiles of hypophyseal LHβ and FSHβ mRNA and of their corresponding gonadal receptors (LH-R, FSH-R) in Xenopus laevis tadpoles during their ontogeny and sexual differentiation. The first significant elevation of LHβ and FSHβ mRNA was observed at late premetamorphosis. A clear raise of LHβ mRNA was present during prometamorphic stages especially in males, while the LH-R only slowly increased during ontogeny with highest levels during metamorphic climax. In contrast, FSHβ mRNA expression only slightly increased during ontogeny, however in both sexes the FSH-R mRNA was considerably elevated at prometamorphosis and further at metamorphic climax. Our results suggest that LHβ and LH-R mRNA expression might be involved in initial maturation events of gametes, at least in males, while the gradually increase of FSH-R mRNA coincided with the advancing process of gamete maturation in both sexes. The present study provides for the first time evidence based on expression of gonadotropins and their corresponding gonadal receptors that the hypothalamus–pituitary–gonad axis evolves already at early stages of ontogeny and sexual differentiation in amphibians.

Ovarian Follicular Dynamics: A Review with Emphasis on the Bovine Species. Part I: Folliculogenesis and Pre‐antral Follicle Development

Recent scientific research into pre-antral follicular dynamics has resulted in the discovery of a wide range of hormones and local factors that influence primordial follicle activation and contribute to follicular development. The putative role of several of these mediators in the follicle growth process has been elucidated by genetic and molecular investigations. Crucial questions, such as the mechanism for primordial follicle initiation and the interplay between oocyte and granulosa cells in this process, remain however unresolved. This review article commences with a description of the embryogenesis of the ovary and follicles. Next, the different stages in the development from primordial to pre-antral follicle are discussed. Thereafter, a short overview of the various in vitro models for the study of follicular dynamics is presented. Finally, an in-depth discussion of pre-antral follicle development engages in the current hypotheses regarding primordial follicle activation, and the role of gonadotrophins and angiogenesis.

Peritoneal Adhesion and Angiogenesis in Ovarian Carcinoma Are Inversely Regulated by Hyaluronan: The Role of Gonadotropins

Ovarian carcinoma is the leading cause of death among gynecologic cancers. Although transformation of the outer ovarian epithelium was linked with ovulation, the disease is significantly more prevalent and severe in postmenopausal women. We postulated that menopause could augment ovarian cancer progression through the effects of gonadotropins on multifocal seeding to the mesothelial layer lining the peritoneum. This seeding is mediated by integrins as well as by CD44 interaction with hyaluronan (HA). Here, we report the effect of gonadotropins on HA synthesis and degradation and on peritoneal adhesion. A significant concentration- and time-dependent induction in expression levels of HA synthases (HASs) and hyaluronidases (Hyals) was observed in vitro on stimulation of human epithelial ovarian carcinoma cells by gonadotropins. Hormonal regulation of HA-mediated adhesion was manifested in vivo as well, by fluorescence microscopy of stained MLS multicellular tumor spheroids. The number of spheroids adhered to the mesothelium of ovariectomized CD-1 nude mice 9.5 hours after intraperitoneal insertion was significantly higher than in nonovariectomized mice. Inhibition of HA synthesis by 6-diazo-5-oxo-1-norleucine (DON) both in spheroids and ovariectomized mice significantly reduced the number of adhered spheroids. Thus, the change in the hormonal environment during menopause assists in HA-dependent adherence of ovarian cancer spheroids onto the peritoneum. However, HA is antiangiogenic and it can significantly suppress tumor progression. Accordingly, angiogenesis of the adhered spheroids was significantly elevated in DON-treated tumors. These results can explain the selective pressure that can lead to simultaneously increased tumor expression of both HASs and Hyals.

Gonadotrophic Hormones – An Overview of their Involvement in Gonadal and Extragonadal Tumours

The concept of the direct involvement of gonadotrophins in tumorigenesis has been around for a long time. First, because the gonads are direct targets of gonadotrophin action, their tumours have been proposed to be gonadotrophin-dependent. Second, the recent findings of gonadotrophin receptors in extragonadal tissues has prompted the hypothesis that some extragonadal tumours (e.g. breast, uterus, prostate, pituitary and adrenal) could also be under the direct regulatory action of gonadotrophins. However, although supported by numerousin vitroexperiments and experimental animal models, the clinical evidence for a direct tumorigenic role of gonadotrophins remains weak. The purpose of this brief review is to present a critical evaluation of current information, both clinical and experimental, about the involvement of gonadotrophins in the induction and growth of gonadal and extragonadal tumours.

Role of gonadotropins on regulation of micro RNAs (miRNAs) and P53 during folliculogenesis

OBJECTIVE: We have previously shown that miRNAs may play a role as regulators of folliculogenesis and oocyte maturation. In this study, we investigate the impact of gonadotropins on miRNAs and p53 expression during follicular maturation in vitro. DESIGN: Expressions of miR-125b, miR-34b and p53 in granulosa cells of follicles matured with long or short-term gonadotropins stimulation were assessed. MATERIALS AND METHODS: Preantral follicles isolated from 14-day-old BDF1 mice were individually cultured in microdroplets, and then stimulated with FSH/LH for 11 days (Group A), or for 6 days (Group B). In-vitro ovulation was induced by hCG/EGF on Day 11. Expression profiles of miR-125b, miR34b and p53 in granulosa cells extracted from Graffian follicles of Groups A and B, and from Graffian follicles before and after ovulation in Group A were assessed by real-time qRT-PCR. RESULTS: Withdrawal of hormones during stimulation resulted in a marked reduction of Graffian formation and ovulation rates (57% and 48% in Group A; 21% and 1.9% in Group B, respectively) and exhibited a dramatic change in gene expression profile of granulosa cells in Graffian follicles. Granulosa cell expression of miR-125b was significantly down-regulated (34%), while expression of miR-34b (176%) and p53 (263%) were up-regulated in Group B. After ovulation, the follicular expression of miR-125b was down-regulated to 29% and p53 was up-regulated to 185% while miR-34 remained unchanged. CONCLUSIONS: Gonadotropins appeared to regulate the expression of cell proliferation genes (miR-125b) and apoptosis-related genes (miR-34b and p53) in granulosa cells during folliculogenesis. Withdrawal of gonadotropins during in-vitro stimulation significantly down-regulated miR-125b with subsequent elevation of p53 mRNA expression. This elevation could be associated with cell apoptosis and follicular atresia. The observed up-regulation of p53 by hCG/EGF in Group A suggests that p53 may be involved in granulosa cell differentiation and possibly luteinization.