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PubMed1995

[Recombinant FSH. Characteristics and therapeutical values].

Pituitary glycoproteins LH and FSH control gonadal physiology. Modern techniques of molecular biology have allowed the synthesis of human recombinant hormones (rec). In particular rec-FSH is now available for clinical use. This pure preparation of FSH, is available in unlimited quantity. The pharmacokinetics of the rec-hormone is identical to its natural counterpart. The use of rec-FSH in hypophysectomized women has allowed the reconfirmation of the "Two-cell--Two gonadotropins" theory. Indeed rec-FSH induces follicular growth without rise in Estradiol secretion. Clinical applications of rec-FSH and LH include ovarian stimulation, as well as, induction of spermatogenesis in men with hypogonadotrophic hypogonadism. Overall, these pure gonadotropins have the critical advantage of purity, reproducibility and in addition they allow easy modulation of the FSH/LH ratio.

Animals↗
PubMed1994

[Efficacy and endocrinological analysis of combined buserelin-pure FSH-hCG therapy in patients with polycystic ovary syndrome].

The study was designed to investigate the efficacy of combined buserelin-pure FSH-hCG therapy in patients with polycystic ovary syndrome (PCO) and to analyse its underlying hormonal changes. Buserelin was intranasally administered daily to 13 patients (35 cycles) for 4-10 weeks with a mean week of 5.4 +/- 1.6 (SD), followed by concomitant pure FSH administration. Out of the 13 patients, 8 were the ones who had experienced ovarian hyperstimulation syndrome (OHSS) using pure FSH alone. Hormonal analyses were performed on these 8 patients. This combined regimen resulted in a 97.1% ovulation rate (34/35) and a 37.1% incidence rate of OHSS (13/35). In 4 out of the 8 patients who had experienced OHSS, no OHSS was observed in the first cycle of the therapy. Eight patients became pregnant with the therapy. Excluding one patient whose husband had oligozoospermia, 7 patients conceived at the first cycle of the therapy and the pregnancy rate per cycle was 25.0% (7/28). No abortion or multiple pregnancy was observed in any of the 8 cases. Pretreatment with buserelin resulted in significantly decreased serum LH, FSH, LH/FSH ratio, estradiol (E2), testosterone (T), and androstenedione (ASD). Serum E2, T and ASD levels at both preovulatory and midluteal phases were significantly lower with the combined regimen than with pure FSH alone. These results indicate that this combined regimen improves the characteristic endocrine profile of PCO and enables pure FSH to achieve ovulation regularly with a high pregnancy rate, although it does not always inhibit OHSS.

Adult↗
PubMedDate not supplied

Follicle-stimulating hormone (FSH) escape during chronic gonadotropin-releasing hormone (GnRH) agonist and testosterone treatment.

Observations that serum follicle-stimulating hormone (FSH) levels begin to rise after initial suppression during chronic gonadotropin-releasing hormone (GnRH) agonist treatment of men with prostate cancer had led to speculation that FSH escape might in part account for the failure of GnRH agonist analogs to completely suppress spermatogenesis in normal eugonadal men. However, previous studies in healthy young men failed to report FSH escape during GnRH agonist treatment for up to 16 weeks. We considered the possibility that this may have been due to the insensitivity of the FSH assays. Accordingly, using highly sensitive and specific two-site directed fluorometric assays and a sustained-release GnRH agonist formulation, we reexamined the issue of whether serum FSH levels rise after initial suppression during chronic GnRH agonist treatment. Two groups of healthy normal men, 19-50 years of age, received 7.5 mg of a long-acting GnRH agonist microcapsule formulation (Lupron Depot; TAP Pharmaceutical Company, North Chicago, Illinois) on days 1 and 30. In addition, the subjects received either 4 or 8 mg/day testosterone replacement by means of a testosterone microcapsule injected intramuscularly on day 1. Serum luteinizing hormone (LH) and FSH levels were measured by sensitive and specific two-site directed fluorometric assays on multiple occasions during the 3-week control period and the 9-week treatment period. Serum LH levels declined to a nadir between 2 and 4 weeks and stayed suppressed throughout the remainder of the treatment period in both the 4- and 8-mg testosterone groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1995

[A randomized, controlled trial comparing follicle stimulating hormone (FSH) to human menopausal gonadotropin (hMG) in fertilization in vitro].

The adverse effect of raised luteinizing hormone (LH) concentrations on reproductive outcome suggests that exogenous LH administration for ovarian stimulation may not be desirable. The aim of this study was to compare the clinical pregnancy rates between follicle stimulating hormone (FSH) and human menopausal gonadotrophin (HMG) used in in-vitro fertilization (IVF) cycles. A total of 232 infertile patients, with a mean duration of infertility of 67.1 +/- 32.9 months, were selected for IVF (female age < 38 years, FSH < 15 IU/l, and total motile sperm count > 5 x 10(6). A short (flare-up) protocol with daily leuprolide acetate was followed randomly from day 3 with FSH (n = 115) or human menopausal gonadotrophin (HMG; n = 117), at an initial dose of two ampoules per day. A maximum of three embryos was transferred, and the luteal phase was supported with four doses of HCG (2,500 IU). No differences were observed between the two groups in any of the cycle response variables except fertilization rates per oocyte and per patient, both of which were significantly higher with FSH. Clinical pregnancy rates per cycle initiated, per oocyte retrieval and per embryo transfer were 19.1, 21.0 and 22.7% respectively for FSH, and 12.0, 12.8 and 15.4% respectively for HMG. Whilst these differences were not statistically significant, the results of this interim analysis suggest that HMG may be associated with a lower clinical pregnancy rate than FSH.

Adult↗
PubMedDate not supplied

The effects of recombinant FSH on testosterone-induced spermatogenesis in gonadotrophin-deficient (hpg) mice.

In order to clarify the mechanism of synergism between follicle-stimulatory hormone(FSH) and testosterone (T) in the hormonal regulation of spermatogenesis, we studied the effects of recombinant human FSH (rhFSH) on the induction of spermatogenesis by testosterone in congenitally gonadotropin-deficient hpg mice. Weanling (day 21) homozygous hpg mice were administered daily subcutaneous injections of 1, 5, or 10 IU of rhFSH alone or in combination with a subdermal testosterone implant until day 70 of age. Spermatogenesis was quantitated by stereological estimation of germ cell populations and Sertoli cells as well as measurement of diameter of seminiferous tubules and their lumina in perfusion-fixed testes and by counting homogenization-resistant condensed testicular spermatids. Recombinant human FSH alone increased the absolute numbers of spermatogonia (x3.5-fold) and spermatocytes (x3-fold) compared with untreated hpg mice but did not significantly increase Sertoli cell numbers or form any condensed spermatids or tubular lumen. Relative to Sertoli cell numbers, rhFSH alone increased populations of spermatogonia (x2-fold) and spermatocytes (x2-fold). The addition of T to rhFSH further increased the absolute numbers of spermatogonia (x5-fold) and spermatocytes (x3.5-fold) compared with untreated hpg mice as well as increasing tubular diameter and forming tubular lumina. In addition administration of T allowed the completion of spermatogenesis in the presence of intratesticular T levels that were approximately 2% of non-hpg controls. All effects of the FSH + T combination were however no greater than the effects of the equivalent dose of T alone. The present study therefore indicates that rhFSH alone increases proliferation of premeiotic spermatogenic cells but has no effect on the completion of spermiogenesis or on Sertoli cell maturation. Furthermore we were unable to identify any additive effects of FSH with T in the hormonal regulation of spermatogenesis in the hpg mouse. This suggests that FSH or T both may stimulate initial spermatogenic development, but only T can complete spermiogenesis.

Animals↗
PubMed1977

[Growth hormone, prolactin, LH and FSH secretion in tissue culture of pituitary adenomas (author's transl)].

We have performed tissue culture of pituitary adenomas (5 acromegalies, 7 non-acromegalies) and measured GH, PRL, LH and FSH in the media of tissue culture by radioimmunoassay (RIA) for 30-40 days. In addition, GH, PRL, LH and FSH were measured in the plasma and tumor tissue by RIA. GH concentration in the culture media was markedly high in all cases with acromegaly and high also in 2 out of 7 non-acromegaly cases (case 6 and 7). GH concentration in the media rapidly decreased almost as a straight line on a semilogarithmic scale until the 30-40 in day, when GH level became less than 10 ng/ml. PRL concentration in the culture media was high in 5 patients (3 acromegalies, 2 non-acromegalies). PRL concentration regressed more slowly than GH, but still remained higher than 10 ng/ml in all of the patients on the 25-40th day when the studies were discontinued. LH and FSH concentrations in the culture media was high in two patients. One patient was a pituitary adenoma secreting LH and FSH, and another case was a false high secreting one, possibly contaminated with gonadotropin in normal pituitary gland at surgical operation. LH and FSH concentrations rapidly decreased as that of GH. A rare case of primary LH and FSH secreting pituitary adenoma was demonstrated.

Acromegaly↗
PubMed1995

Endothelins inhibit FSH-mediated function via ETA receptors in cultured human granulosa-lutein cells.

Endothelins (ETs) are a family of vasoactive peptides involved in granulosa and luteal cell function in some animal species. However, the potential relevance of ETs in ovarian physiology remains unclear, and the direct action of the peptides in the human ovary has not been studied to date. Experiments were conducted to determine whether ET-1 and ET-3 could regulate the follicle stimulating hormone (FSH)-induced cell response in human granulosa-lutein cells in culture. The FSH-mediated cell rounding process was used as an indicator of cell response, as previously described (Lawrence et al., 1979). Forskolin, cholera toxin, 8-Br-cyclic AMP, isobutylmethylxanthine (IBMX), rolipram and FSH all stimulated similar cell morphological changes, indicating that the cell rounding process was mediated by cyclic AMP. Although ET-1 and ET-3 alone failed to alter cell shape, the FSH-induced cell response was totally inhibited by treatment with ET-1 (10(-10) mol/l) and ET-3 (10(-7) mol/l). In addition, treatment of the cells with BQ123, an antagonist of ET binding on ETA receptor subtype, totally prevented the inhibitory effects of ET-1 and ET-3 on the FSH-induced response. The data presented here show that human granulosalutein cells are a site of ET reception and action. Endothelins inhibit cyclic AMP-dependent FSH-mediated function and the ET(A) receptor participates in this effect.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1991

Sulfhydryl groups are involved in the interaction of FSH with its receptor.

FSH has recently been reported to possess thioredoxin-like activity, presumably explained by the homology between a region of FSH-beta subunit and the active site of thioredoxin. The homologous sequence lies within a receptor binding region, which suggests a possible role for sulfhydryl groups in the formation of an active hormone-receptor complex and subsequent signal transduction. In order to determine the relevance of sulfhydryl groups on FSH-receptor interaction, we studied the effect of N-ethylmaleimide (NEM) and glutathione on FSH binding. The results indicate that free sulfhydryl groups, probably derived from the FSH receptor, are involved in ligand-receptor interaction.

Animals↗
PubMed1991

FSH- and TSH-binding cells in the ovary of the developing chick embryo.

The numerical density (Nv) and location of thyroid stimulating hormone (TSH)- and follicle stimulating hormone (FSH)-binding cells in the left ovary of the chick embryo were determined on Days 6.5 through 19.5 of incubation. From Days 6.5 to 11.5, TSH- and FSH-positive cells were located in the medullary cords and there was no statistically significant difference in the Nv of these two cellular populations. However, beginning on Day 12.5 and continuing through Day 19.5, TSH and FSH were bound principally to cells of the cortical cords and there was a significant difference (P less than 0.05) between the Nv of TSH-positive and FSH-positive cells. Results are discussed in terms of a prevalent hypothesis that throughout chick embryo development the left ovary lacks specific receptors for TSH and FSH.

Animals↗
PubMed2004

Characterization of biologically active bovine pituitary FSH purified by immunoaffinity chromatography using a monoclonal antibody.

A substantial amount of highly purified, biologically active bovine FSH was isolated from pituitary extracts by immunoaffinity chromatography based on a novel anti-bovine FSH beta-subunit monoclonal antibody. The biological activity was assessed in vitro using a steroidogenic granulosa cell line constitutively expressing the FSH receptor. Amino acid analysis, N-terminal amino acid sequencing, and peptide mass mapping demonstrated that primary structure modifications do not contribute to the heterogeneity of bovine FSH. The monosaccharide composition of the N-linked oligosaccharides was quantified and remarkably two distinct forms of sialic acids, N-acetyl- and N-glycolyl-neuraminic acids were found. In conclusion, we showed that isoform differences in bovine FSH is likely due only to sugar chain heterogeneity, and we give the first evidence that two substituted sialic acids contribute to the diversity of mammalian glycoprotein hormone isoforms.

Animals↗
PubMed2006

FSH receptor gene polymorphisms in fertile and infertile Italian men.

In women, single nucleotide polymorphisms (SNP) of the FSH receptor (FSHR) gene influence FSH concentrations and the sensitivity of the FSHR to FSH in vivo. In contrast, the significance of FSHR R gene SNP in the male is poorly understood. To this aim, the possible role of three FSHR SNP was evaluated in male infertility. SNP in exon 10 (codon 307 and 680) and in the core promoter region (at position -29) of the FSHR gene were analysed by polymerase chain reaction-restriction fragment length polymorphism technique in 150 men representative of the general population, 107 proven fathers, 92 normozoospermic controls, and 215 infertile patients classified according to sperm parameters (38 azoospermia, 53 severe oligozoospermia, 48 moderate oligozoospermia, and 76 slight oligozoospermia). Reproductive hormones were measured in infertile males and normozoospermic controls. No significant difference was found in allelic variants frequency and genotype distribution between each category of subjects when analysing the FSHR exon 10 SNP alone and in combination with the SNP at position -29. Serum FSH concentrations and other andrological parameters did not differ between subjects with different genotype within each group. The data showed that in the Italian population, FSHR genotypes have no influence on FSH concentrations both in normal and infertile males and do not associate with spermatogenetic impairment.

Alleles↗
PubMedDate not supplied

Intact follicle culture: what it can tell us about the roles of FSH glycoforms during follicle development.

An important limiting factor in assisted reproduction treatment success rates is oocyte quality. In spite of improved results through several important innovations, the pregnancy rate per collected oocyte remains far too low. In order to improve this situation, it is necessary to learn more about fundamental factors modulating follicular development patterns. FSH is known to be the driving force for follicle development, but it is not yet understood how its multifarious functions are controlled and modulated. Evidence is accumulating that FSH glycoforms may be the key to this mystery. Intact follicle culture is a useful tool for the clarification of the actions of the different isoforms because the follicle unit is maintained and allowed to develop through several critical stages. Additionally important is the availability of the oocyte for functional evaluation. Because of these features, relationships can be uncovered that are not revealed with single cell test systems. The results so far obtained with this system suggest that follicle development pattern and oocyte quality is strongly influenced by FSH glycoform range, and that the requirements of the follicle may shift during progress through different stages of development. More studies are required, but these findings already suggest that the physiological shifts of circulating FSH glycoforms may indeed be important, and that attention should be paid to the glycoform distribution of exogenously applied FSH.

Animals↗
PubMed1982

Effects of induced hypoprolactinaemia in the ram: plasma gonadotrophin levels, LH and FSH receptors and histology of the testis.

In order to investigate the effects of induced hypoprolactinaemia on gonadotrophin levels and testicular function in the ram, CB 154 was administered either in summer under natural photoperiod or in winter combined with a light regime stimulating prolactin release. Under natural photoperiod in summer, plasma FSH levels increased from 2.5 to 4 ng/ml in controls and from 2.5 to 8 ng/ml in CB 154-treated rams. Control and treated groups differed significantly (P less than 0.05) from the second week of treatment onwards. In the animals photostimulated in winter, treatment with CB 154 did not change plasma FSH levels; no effect of CB 154 on plasma LH levels was detected in either experiment. Treatment with CB 154 in summer led to an increase in the volumes of peritubular blood vessels and interstitial tissue. The number of Leydig cells per testis was significantly (P less than 0.05) increased in CB 154-treated rams. However, at the time of measurement, there was no change in the structure of the seminiferous tubules or in the production of germ cells. In winter in photostimulated animals, no effect of treatment with CB 154 was detected either on the structure of intestitial and tubular compartments or on spermatogenesis. The change in prolactin levels affected neither the number of LH receptors, expressed per Leydig cell, nor the number of FSH receptors, expressed per Sertoli cell, in the experiments. It is concluded that induced hypoprolactinaemia can modify the release of FSH in the ram and change the structure of the intertubular tissue but that it does not affect the number of LH and FSH receptors or impair spermatogenesis.

Animals↗
PubMedDate not supplied

Screening of FSH receptor gene mutation (C566T) in azoospermic men in Japan.

To study the genetic mutation, which could cause problems in spermatogenesis, we screened the point mutations of the FSH receptor gene (C566T) in idiopathic azoospermic men in Japan. We performed mutational analysis of the FSH receptor in 54 Japanese patients diagnosed for azoospermia with Sertoli cell-only (SCO) syndrome (n = 33), hypospermatogenesis (n = 11) and maturation arrest (n = 10). For mutation screening of the FSH receptor, polymerase chain reaction (PCR) amplification from genomic DNA with flanking intronic primers were used. On BsmI digestion, all patients demonstrated homozygous, normal exon 7 alleles with 51 and 27 bp fragments. The absence of any 78 bp fragments demonstrated that no heterozygous or homozygous mutant alleles were present in any patients. None of the 54 patients showed a C566T FSH receptor mutation. We could not confirm that the genomic mutation of the FSH receptor (C566T) is a common cause in Japanese azoospermic patients.

Adult↗
PubMed2002

Genetic and functional analyses of polymorphisms in the human FSH receptor gene.

To determine the influence of FSH receptor variants Thr307-Asn680 (TN) and Ala307-Ser680 (AS) on ovarian function, we investigated the frequency of these gene polymorphisms by using restriction fragment length polymorphism analysis and observed their effects on clinical manifestations. In a population of 522 Japanese women, the overall frequency of TN/TN (NN), TN/AS (NS), and AS/AS (SS) was 41.0, 46.9 and 12.1% respectively. In polycystic ovary patients, the NS population was significantly larger when compared with the spontaneously ovulating group (66.7 versus 43.5%, P < 0.05). In the SS group, a significantly higher (46%) basal level of serum FSH was observed as compared with that in the NS group (P < 0.05). A higher dose of the exogenous gonadotrophin was required to achieve ovulation induction in the SS group as compared with the NS group (P < 0.05). At the time of hCG administration, estradiol levels per oocyte retrieved for IVF in the SS group were significantly lower as compared with the levels in the NS and NN groups (P < 0.05). There were no significant differences in FSH-stimulated cAMP production and PI turnover as well as ligand-binding affinity between the two receptor isoforms when overexpressed in transfected 293T cells. These results suggest that although FSH receptor polymorphisms have no discernible effect on FSH receptor function in vitro, there are associations between the genotype and some aspects of patient status.

Alleles↗
PubMed2006

Targeted and reversible disruption of the blood-testis barrier by an FSH mutant-occludin peptide conjugate.

The blood-testis barrier (BTB) is one of the tightest blood-tissue barriers in mammals. As such, it poses a challenge to deliver any drugs to the seminiferous epithelium of the testis, such as a nonhormonal male contraceptive. To circumvent this problem, a genetically engineered follicle-stimulating hormone (FSH) mutant protein was produced in Spodoptera furgiperda (Sf)-9 insect cells to serve as a testis-specific carrier. Subsequently, a 22-amino acid peptide corresponding to the second extracellular loop of occludin, which was known to disrupt BTB integrity in vivo, was inserted to the FSH mutant by polymerase chain reaction (PCR), as well as chemical cross-linking. This molecule was found to have negligible hormonal activity but was still capable of binding to FSH receptors, which are restricted to Sertoli cells in mammals. When this FSH mutant-occludin peptide conjugate was administered to adult rats at 40 microg/adult rat (approximately 300 gm b.w.) via intraperitoneally (i.p.) injection, it induced transient and reversible disruption of the BTB, while at 150 microg/rat, it induced partial germ cell loss from the testis, particularly elongating/elongate spermatids. Most importantly, this effect was limited to the BTB without compromising the TJ-barrier integrity or cell adhesion in epithelia of other organs, such as kidney, liver, and small intestine. In summary, the use of an FSH mutant-occludin peptide conjugate is a feasible nanodevice to transiently compromise the BTB.

Animals↗
PubMed2006

Identification of differential gene expression in in vitro FSH treated pig granulosa cells using suppression subtractive hybridization.

FSH, which binds to specific receptors on granulosa cells in mammals, plays a key role in folliculogenesis. Its biological activity involves stimulation of intercellular communication and upregulation of steroidogenesis, but the entire spectrum of the genes regulated by FSH has yet to be fully characterized. In order to find new regulated transcripts, however rare, we have used a Suppression Subtractive Hybridization approach (SSH) on pig granulosa cells in primary culture treated or not with FSH. Two SSH libraries were generated and 76 clones were sequenced after selection by differential screening. Sixty four different sequences were identified, including 3 novel sequences. Experiments demonstrated the presence of 25 regulated transcripts.A gene ontology analysis of these 25 genes revealed (1) catalytic; (2) transport; (3) signal transducer; (4) binding; (5) anti-oxidant and (6) structural activities. These findings may deepen our understanding of FSH's effects. Particularly, they suggest that FSH is involved in the modulation of peroxidase activity and remodelling of chromatin.

Animals↗
PubMed1997

Immunization of male bonnet monkeys (M. radiata) with a recombinant FSH receptor preparation affects testicular function and fertility.

Immunization of proven fertile adult male monkeys (n=3) with a recombinant FSH receptor protein preparation (oFSHR-P) (representing amino acids 1-134 of the extracellular domain of the receptor Mr approximately 15KDa) resulted in production of receptor blocking antibodies. The ability of the antibody to bind a particulate FSH receptor preparation and receptors in intact granulosa cells was markedly (by 30-80%) inhibited by FSH. Serum T levels and LH receptor function following immunization remained unchanged. The immunized monkeys showed a 50% reduction (p<0.001) in transformation of spermatogonia(2C) to primary spermatocytes (4C) as determined by flow cytometry and the 4C:2C ratio showed a correlative change (R 0.81, p<0.0007) with reduction in fertility index (sperm counts X motility score). Breeding studies indicated that monkeys became infertile between 242-368 days of immunization when the fertility index was in the range of 123+/-76 to 354+/-42 (compared to a value of 1602+/-384 on day 0). As the effects observed are near identical to that seen following immunization with FSH it is suggestive that oFSHR-P can substitute for FSH in the development of a contraceptive vaccine.

Animals↗