FSH Receptor Genotype Research Articles

Luteal phase stimulation in double ovarian stimulation cycles is not affected by the follicle-stimulating hormone (FSH) receptor genotype: double ovarian stimulation is beneficial independently of the genotype at position 680 of the follicle-stimulating hormone receptor.

To determine whether follicle-stimulating hormone receptor (FSHR) genotype influences the outcome of ovarian stimulation treatment in luteal phase. A total of 299 patients were included in a retrospective study between July 2017 and December 2021. These patients carried out a double stimulation protocol and the variant Asn680Ser (rs6166; c.2039A>G) of FSH receptor was genotyped either as part of the pre-treatment fertility tests or for the current study. Patients undergoing a double stimulation treatment who could not be genotyped were excluded from this analysis. The results obtained from ovarian stimulation in luteal phase were better than those obtained in conventional follicular phase. Statistically significant differences ( P < 0.001) were found in the number of retrieved oocytes (5.47 vs. 4.18), retrieved MII (4.52 vs. 3.29) and fertilised oocytes (3.81 vs. 2.20). Furthermore, these differences remained regardless of the FSH receptor genotype for the 680 position in all groups ( P < 0.05). In addition, stimulation in luteal phase lasts longer and requires more gonadotropins than in follicular phase. This is especially noteworthy in patients with Ser/Ser genotype, who required a slightly higher dose of gonadotropins compared to other genotypes in luteal phase, as previously observed in the follicular phase for this genotype. No significant differences in age, anti-Müllerian hormone levels, antral follicle count, BMI and type of trigger used in luteal phase were observed among groups of patients with different FSH receptor genotypes. All patients undergoing IVF seem to benefit from luteal phase ovarian stimulation, regardless of their FSHR genotype.

Spontaneous and iatrogenic ovarian hyperstimulation syndrome in the absence of FSHR mutations: a case report of two unexpected cases

BackgroundOvarian hyperstimulation syndrome (OHSS) is a complication of controlled ovarian hyperstimulation (COH). It is a potentially life-threatening condition that usually occurs either after human chorionic gonadotropins (hCG) administration in susceptible patients or as a result of an implanting pregnancy, regardless of whether it was achieved by natural conception or infertility treatments. Despite many years of clinical experience regarding the adoption of preventive measures and the identification of patients at high risk, the pathophysiology of OHSS is poorly understood and no reliable predictive risk factors have been identified.Cases presentationWe report about two unexpected cases of OHSS following infertility treatments, occurring after freeze-all strategy with embryo cryopreservation approaches. The first case developed spontaneous OHSS (sOHSS), despite efforts to prevent its manifestation by a segmentation approach, including frozen embryo replacement cycle. The second case developed a late form of iatrogenic OHSS (iOHSS), even though the absence of any risk factors. No mutations in the follicle-stimulating hormone (FSH) receptor (FSHR)-encoding gene were detected, suggesting that the high levels of hCG due to the twin implanting pregnancies could be the only triggering factor of OHSS outbreak.ConclusionFreeze-all strategy with embryo cryopreservation cannot entirely prevent the development of OHSS, which may occur in its spontaneous form independently from the FSHR genotype. Although OHSS remains a rare event, all infertile patients requiring ovulation induction or controlled ovarian stimulation (COS) may be at potential risk of OHSS, either in the presence or in the absence of risk factors. We suggest closely monitoring cases of pregnancy following infertility treatments in order to provide early diagnosis and adopt the conservative management.

The Ala307Thr polymorphism of the follicle-stimulating hormone receptor (FSHR) gene is associated with the dose of recombinant FSH received during IVF/ICSI treatment.

Follicle-stimulating hormone (FSH) is essential for folliculogenesis, acting through the follicle-stimulating hormone receptor (FSHR) that is present on the membrane of granulosa cells. Polymorphisms in the FSHR gene may lead to an altered pattern of receptor expression on the cell surface or to changes in affinity for FSH. The aim of this prospective study was to detect any association between the follicle-stimulating hormone receptor (FSHR) gene Ala307Thr polymorphism (rs6165) and ovarian reserve, ovarian response or clinical results in IVF/ICSI treatment. This prospective cohort study included 450 women who underwent IVF/ICSI cycles. DNA was extracted from peripheral blood, and the Ala307Thr FSHR polymorphism (rs6165) was genotyped using the TaqMan SNP genotyping assay. Participants were divided into three groups according to their Ala307Thr FSHR genotype: Thr/Thr (n:141), Thr/Ala (n=213) and Ala/Ala (n=96). The results were tested for associations with age, anti-Mullerian hormone (AMH) levels, antral follicle count (AFC), total dose of r-FSH, follicle size, number of retrieved oocytes, and clinical outcome of IVF/ICSI cycles. The statistical analyses were performed using Fisher's exact test and the Kruskal‒Wallis test. An association between the genotype of the FSHR (Ala307Thr) polymorphism and the dose of r-FSH was observed. Patients with the Ala/Ala genotype received a higher r-FSH dose than patients with the Ala/Thr (p=0.0002) and Thr/Thr (p=0.02) genotypes. No other correlation was observed. The Ala/Ala genotype was associated with the use of higher doses of recombinant FSH (r-FSH), suggesting that homozygosis of this allelic variant (Ala) provides lower sensitivity to r-FSH.

Polymorphism of the follicle stimulating hormone receptor does not impact reproductive performance or in-vitro embryo production in beef heifers

Assisted reproductive technologies are used to propagate desirable genetics in a shortened timeframe. Selected females undergo ovarian stimulation with the use of follicle stimulating hormone (FSH) to increase embryo recovery for subsequent transfer programs. The FSH receptor (FSHR) c.337 C > G variant was reported to have a reduction in viable embryo numbers in an ovarian stimulation protocol. We, therefore, hypothesized that FSHR c.337 C > G would result in reduced in-vitro blastocyst development. Beef heifers were genotyped and selected based on the c.337 C > G FSHR genotype (CC, CG, GG; n = 15–16/genotype). Estrus was synchronized with a Select Synch protocol and heifers were slaughtered 5 days after induced ovulation. Anterior pituitaries, serum and reproductive tracts were collected at slaughter for analysis. Cumulus oocyte complexes (COCs) were collected and pooled within genotype for in-vitro fertilization (IVF) and subsequent blastocyst development. No differences were observed in carcass weights, anterior pituitary weights, serum progesterone, corpus lutea weight, surface follicle counts, histological follicle counts or follicular fluid estradiol concentration (P > 0.1) due to FSHR genotype. Differences were observed for ovulation rates in the GG FSHR genotype group (P < 0.01). However, cleavage and blastocyst rates were not affected due to FSHR genotype (P > 0.1), following standard IVF protocols. The FSHR variant does not influence antral follicle counts, estradiol production, or in-vitro blastocyst development in beef heifers. The GG FSHR genotype had an increased ovulation rate, which may indicate a greater potential for twinning, but research with a larger population is warranted to support this hypothesis.

FSH receptor genotype and its influence on the results of donor ovarian stimulation using corifollitropin alfa

Research questionDoes the FSH receptor (FSHR) genotype influence the results of donor ovarian stimulation using corifollitropin alfa? DesignA prospective cohort study was performed including 152 oocyte donor ovarian stimulations: group 1 (n = 80) using a single dose of 150 µg of corifollitropin alpha; and group 2 (n = 72) using in addition to corifollitropin alpha, continued stimulation using recombinant FSH 225 IU daily. Allelic discrimination was used to genotype the FSHR p.N680S polymorphism. Linear regression analysis was performed to study the differences between groups. ResultsNo differences in clinical characteristics between genotypes were reported. Overall, the results of ovarian stimulation were better in oocyte donors with SN and NN genotypes compared with SS in terms of the number of retrieved oocytes (15.78 versus 10.83; P = 0.008) and retrieved metaphase II (MII) oocytes (12.34 versus 9.00; P = 0.032). Corresponding differences were also observed in group 1 for the number of retrieved oocytes (13.83 versus 7.50, P = 0.018) and retrieved MII oocytes (10.24 versus 5.42; P = 0.038). However, in group 2 no significant differences were found for oocytes retrieved (17.55 versus 13.06, P = 0.064) or MII oocytes (14.25 versus 11.39; P = 0.12). ConclusionsThis study suggests that ovarian stimulation protocols with corifollitropin alfa in women with the SS genotypes could be associated with fewer oocytes and MII oocytes retrieved. Despite the fact that corifollitropin alfa has a longer half-life, the results for the SS genotype do not match those for the other genotypes, so other factors must be involved. Therefore, to tailor treatments, it would be advisable to genotype women at p.N680S of the FSHR.

Вклад эндокринной системы в развитие остеопороза у лиц пожилого и старческого возраста (обзор)

Osteoporosis is increasingly found in the elderly and senile, maintaining its enormous medical and social significance. The effect of hormones on bone metabolism is beyond doubt. However, currently the data on the effect of sex hormones on bone tissue prevails. As for the other hormones, sometimes, there are conflicting opinions. The aim of the study: Based on published data, to study the contribution of the endocrine system to the development of osteoporosis in the elderly. Materials and methods: Literature data was analyzed using the following search words: osteoporosis, bone mineral density, FSH, estrogens, testosterone, cortisol, vitamin D, IGF1 for 1998-2020 in computer databases PubMed, Scopus, Medical- Science, Elibrary, Web of Science, Ceeol. Results: Analysis of the literature showed that the increase of levels of thyroid stimulating hormone (TSH) plays an osteoprotective role; the decrease of levels of estrogen, testosterone, insulin-like growth factor 1 (IGF1) and vitamin D, as well as the increase in the levels of cortisol, parathyroid hormone and follicle-stimulating hormone (FSH) contribute to bone loss in the elderly and senile. In addition, the FSH receptor (FSHR) genotype AA rs6166 is associated with low bone mineral density, regardless of estrogen level. A polyclonal antibody with an FSHR-binding sequence against mouse β-subunit of FSH is likely to be an effective tool for reducing bone loss in mice subjected to ovariectomy. Conclusion: A comprehensive assessment of the hormonal profile in the elderly and senile is needed to identify the causes of osteoporosis and the formation of an individual program of medical diagnostic and rehabilitation measures. Currently, there are all prerequisites for the development of new diagnostic and therapeutic interventions for the correction of low bone density.

P–588 Follicle-stimulating hormone receptor genotype and its influence on the results of double ovarian stimulation in IVF cycles

Abstract Study question Does the follicle-stimulating hormone receptor (FSHR) genotype influence the results of the ovarian stimulation treatment in the luteal phase? Summary answer All patients undergoing in-vitro fertilization benefit from luteal phase ovarian stimulation, regardless of their follicle-stimulating hormone receptor genotype. What is known already Previous studies suggest that FSH receptor polymorphism in position 680 influences the response to ovarian stimulation in the luteal phase. It was observed that patients with SS genotype seems to require a higher dose to obtain an optimal ovarian response. Later, it was reported that, in patients with SS genotype, a better performance seems to be obtained by administering highly purified urinary FSH while, in SN patients, better results were obtained with recombinant FSH. In patients with NN genotype, no differences were found. Our aim was to test whether this concept is applicable to ovarian stimulation in the luteal phase. Study design, size, duration One hundred and thirty-four patients were included in a retrospective study between July 2017 and September 2020. In these patients, a double stimulation protocol was carried out and the FSH receptor was genotyped either as part of the pre-treatment fertility tests or for the current study. Patients with a double stimulation treatment who could not be genotyped were excluded from the analysis. Participants/materials, setting, methods To genotype the 680 position of the FSH receptor, a real-time PCR for allelic discrimination was carried out using StepOnePlus™ Real-Time PCR System (Applied Biosystems™. Ref: 4376600). Non-parametic tests were used to study the differences between the groups. They were performed with the software R Statistical Software, version 4.0.3. Main results and the role of chance The results of ovarian stimulation in the luteal phase were better compared to the conventional follicular phase. Statistically significant differences (p &amp;lt; 0.001) were found in the number of retrieved oocytes (5.06 versus 3.51), retrieved MII (4.13 versus 2.91), fertilized oocytes (3.22 versus 1.81) and blastocysts formed (1.79 versus 0.62). Furthermore, these differences remained regardless of the genotype for the 680 position of the FSH receptor in all groups (p &amp;lt; 0.05). In addition, better results were obtained in the luteal phase in patients who have been stimulated with the type of gonadotropin that already had better performance in the follicular phase for its genotype, that is, highly purified urinary FSH in SS patients and recombinant FSH in SN patients, compared to other types of gonadotropin (p &amp;lt; 0.05). We also observed that stimulation in the luteal phase lasts longer and consume more gonadotropins than in the follicular phase. This is especially notable in the case of patients with SS genotype, who required slightly higher consumption of gonadotropins compared to the other genotypes in the luteal phase, as had previously been observed in the follicular phase for this genotype. Limitations, reasons for caution The retrospective study design and the sample size could be a limitation. Furthermore, we cannot determine whether the improvement in luteal phase performance is related to differences in the physiological environment between phases of the cycle or is caused by a possible activation of the ovary from the previous stimulation. Wider implications of the findings: All patients undergoing in-vitro fertilization seems to benefit from luteal phase ovarian stimulation, regardless of their genotype for FSHR. In addition, the pharmacogenetic recommendation when choosing the type of FSH for ovarian stimulation should be the same both in the follicular phase and in the luteal phase. Trial registration number Not applicable

The Role of FSHR SNPs and AMH in Follicular Fluid and Serum in Ovarian Response during COS: A Pilot Study.

Background Several studies have investigated on the polymorphism Ser680Asn of FSHR and its use as a predictive indicator of response to an IVF/ICSI protocol. Furthermore, measurement of AMH in serum and follicular fluid is a useful prognostic indicator for the outcome of an assisted reproduction attempt. The purpose of this study is to examine the FSH receptor Ser680Asn polymorphism in combination with AMH levels in both serum and follicular fluid, on the day of oocyte collection. Materials and Methods A total of 32 women who underwent IVF/ICSI were included. Women were grouped into 2 groups: those who received rFSH (n = 11) and those who received hMG (n = 21). Serum AMH was measured on day 3 of the cycle, and AMH in the follicular fluid on the day of oocyte retrieval; the same day peripheral blood was collected for the genotyping of Ser680Asn. Results No statistical significant difference was found between serum AMH and follicular fluid AMH regarding the FSH receptor genotype for the Ser680Asn polymorphism. Regarding the sAMH/ffAMH ratio in the 3 genotypes, the value was lower in Asn/Asn women than Ser/Ser and Ser/Asn, but no statistical difference was obtained. Women who carry the Ser allele have a higher number of follicles, retrieved oocytes, and mature oocytes than women who do not contain the Ser allele. Women with AMH < 2.22 ng/ml presented lower AMH follicular fluid levels and lower serum AMH/follicular fluid AMH ratio in a statistically significant manner. Concerning the genotype for the polymorphism Ser680Asn of FSHR in relation to AMH levels, no statistically significant differences were found. Conclusions The identification of polymorphisms, such as Ser680Asn of FSHR, along with the determination of endocrine markers in the follicular fluid, such as AMH, could lead at some point, to the personalized therapy setting per woman.

FSHR, a Novel Major Fish Sex-Determining Locus Shows Variable Incomplete Penetrance Across Flathead Grey Mullet Populations

Whole-genome sequence data were produced from a single flathead grey mullet female and assembled into a draft genome sequence, whereas publicly available sequence data were used to obtain a male draft sequence. Two pools, each consisting of 60 unrelated individuals respectively of male and female fish were analysed using Pool-Sequencing. Mapping and analysis of Pool-Seq data against the draft genome(s) revealed >30 loci potentially associated with sex. The most promising of these loci encoding the follicle stimulating hormone receptor (FSHR) and harbouring two missense variants, was genotyped on 245 fish from four Mediterranean populations. Genotype data showed that FSHR represents a previously unknown sex-determining locus. However, the incomplete association pattern between FSHR genotype and sex-phenotype, the variability of such pattern across different natural populations and the presence of other candidate loci suggest a greater complexity underling sex determination in the flathead grey mullet.

FSH Receptor Gene Polymorphism in Indonesian Women with Polycystic Ovarian Syndrome (PCOS)

Polycystic ovary syndrome (PCOS) is a reproductive problem that often occurs among women in reproductive age, but until now, the etiopathogenesis of PCOS is still unclear. One of the alleles for candidate genes associated with the PCOS incident is the FSH receptor (FSHR) gene. This research was aimed at analyzing the genotypes within Asn680Ser FSHR with PCOS pathogenesis. The total research subject was 142 subjects. DNA isolation was used for performing the RFLP analysis toward Asn680Ser FSHR polymorphism. There was no mean value difference of the genotype distribution within Asn680Ser FSHR between the PCOS and non-PCOS groups (p>0.05). There was no difference of the basal FSHR levels based on the Asn680Ser FSHR genotyping variation between the PCOS and the non-PCOS groups (p >0.05). The Asn680Ser FSHR genotype does not determine the individual susceptibility to suffering from PCOS. There was no difference in basal FSH levels between PCOS and non-PCOS groups based on the Asn680Ser FSHR genotype variation. However, there are differences in the frequency distribution pattern of the FSHR genotype at position 680 in the PCOS group compared to the previous research findings in several countries.

Osteoporosis from an Endocrine Perspective: The Role of Hormonal Changes in the Elderly.

Introduction: Osteoporosis is increasingly prevalent in the elderly, with fractures mostly occurring in women and men who are older than 55 and 65 years of age, respectively. The aim of this review was to examine the evidence regarding the influence of hormones on bone metabolism, followed by clinical data of hormonal changes in the elderly, in the attempt to provide possible poorly explored diagnostic and therapeutic candidate targets for the management of primary osteoporosis in the aging population. Material and methods: An extensive Medline search using PubMed, Embase, and Cochrane Library was performed. Results: While the rise in Thyroid-stimulating hormone (TSH) levels has a protective role on bone mass, the decline of estrogen, testosterone, Insulin-like growth factor 1 (IGF1), and vitamin D and the rise of cortisol, parathyroid hormone, and follicle-stimulating hormone (FSH) favor bone loss in the elderly. Particularly, the AA rs6166 FSH receptor (FSHR) genotype, encoding for a more sensitive FSHR than that encoded by the GG one, is associated with low total body mass density (BMD), independently of circulating estrogen. A polyclonal antibody with a FSHR-binding sequence against the β-subunit of murine FSH seems to be effective in ameliorating bone loss in ovariectomized mice. Conclusions: A complete hormonal assessment should be completed for both women and men during bone loss evaluation. Novel possible diagnostic and therapeutic tools might be developed for the management of male and female osteoporosis.

The relationship between FSH receptor polymorphism status and IVF cycle outcome: a retrospective observational study.

What is the association between FSH receptor (FSHR) gene polymorphism at position 680 and live birth in women undergoing IVF and intracytoplasmic sperm injection (ICSI). In this retrospective cohort study, data were collected from the Electronic Patient Database of the VU University Medical Centre, Amsterdam, The Netherlands. Women undergoing their first IVF/ICSI cycle between January 2008 and March 2012, of whom the FSHR genotype was determined, were included. The main outcome was live birth rate. Secondary outcomes were ongoing pregnancy, total number of follicles, oocytes and embryos. The FSHR genotype distribution was as follows: 334 women in the Asn/Asn group (28.2%), 617 in the Asn/Ser group (52.1%) and 234 in the Ser/Ser group (19.7%). Basal FSH concentration was highest in the Ser/Ser group (P = 0.006). The number of oocytes (P = 0.01) and number of embryos (P = 0.02) were lowest in the Ser/Ser group. The Asn/Asn group showed a significantly lower live birth rate. Live birth rates were 21.9% versus 31.1% and 27.6% (P = 0.009), for Asn/Asn, Asn/Ser and Ser/Ser, respectively. Logistic regression analysis, however, showed no significant difference on cumulative live birth rate between the three genotypes either unadjusted or when adjusted for age. The homozygous Ser/Ser genotype of FSHR polymorphism at position 680 is associated with a reduced ovarian response to ovarian stimulation in IVF/ICSI. No difference in cumulative live birth rate was found.

Association of Thr307Ala and Asn680Ser of Follicle-Stimulating Hormone Receptor Gene Polymorphisms with Gonadotropin Administration during Controlled Ovarian Hyperstimulation

Objective: This study is to investigate the effect of different single-nucleotide polymorphisms of follicle-stimulating hormone receptor (FSHR) gene on gonadotropin (Gn) administration dosage during controlled ovarian hyperstimulation (COH) protocol of in vitro fertilization and embryo transfer. Methods: This retrospective study included 184 Chinese infertile women in Center for Reproduction and Genetics of Suzhou Municipal Hospital from June 2012 to 2014. All of the enrolled patients were homogeneous in some basal characteristics, and they all met the eligibility criteria. Blood tests were conducted on day 3 of menstrual cycle or the day of human chorionic gonadotropin administration for hormonal profile analysis and DNA extraction. DNA sequencing was performed for polymorphism analysis. The participants were classified into threonine (Thr)/Thr, Thr/alanine (Ala), and Ala/Ala groups according to genotype at position 307, and asparagine/asparagine (Asn/Asn), Asn/serine (Ser), and Ser/Ser groups according to genotype at position 680. Logistic regression and correlation analyses were performed to identify the effect of these two polymorphisms on Gn consumption. Results: The frequency of Thr307Ala and Asn680Ser distribution was consistent with Hardy–Weinberg equilibrium (P > 0.05). No significant difference was found in age, basal hormone levels for different genotype groups. Logistic regression analysis results revealed that patients with Ser680Ser genotype have a higher risk of requiring a high dose of Gn compared with patients with Asn680Asn genotype, while polymorphism of Thr307 Ala has no such effect. Conclusion: This study suggested that FSHR genotype Asn680Ser would be helpful in determining the dosage of Gn in COH; patients with Ser680Ser genotype may require higher dose of Gn.

Treatment with human, recombinant FSH improves sperm DNA fragmentation in idiopathic infertile men depending on the FSH receptor polymorphism p.N680S: a pharmacogenetic study

Does the sperm DNA fragmentation index (DFI) improve depending on the FSH receptor (FSHR) genotype as assessed by the nonsynonymous polymorphisms rs6166 (p.N680S) after 3 months of recombinant FSH treatment in men with idiopathic infertility? FSH treatment significantly improves sperm DFI only in idiopathic infertile men with the p.N680S homozygous N FSHR. FSH, fundamental for spermatogenesis, is empirically used to treat male idiopathic infertility and several studies suggest that DFI could be a candidate predictor of response to FSH treatment, in terms of probability to conceive. Furthermore, it is known that the FSHR single nucleotide polymorphism (SNP) rs6166 (p.N680S) influences ovarian response in women and testicular volume in men. A multicenter, longitudinal, prospective, open-label, two-arm clinical trial was performed. Subjects enrolled were idiopathic infertile men who received 150 IU recombinant human FSH s.c. every other day for 12 weeks and were followed-up for a further 12 weeks after FSH withdrawal. Patients were evaluated at baseline, at the end of treatment and at the end of follow-up. Eighty-nine men with idiopathic infertility carrier of the FSHR p.N680S homozygous N or S genotype, FSH ≤ 8 IU/l and DFI >15%, were enrolled. A total of 66 patients had DFI analysis completed on at least two visits. DFI was evaluated in one laboratory by TUNEL/PI (propidium iodide) assay coupled to flow cytometry, resolving two different fractions of sperm, namely the 'brighter' and 'dimmer' sperm DFI fractions. Thirty-eight men (57.6%) were carriers of the p.N680S homozygous N and 28 (42.4%) of the homozygous S FSHR. Sperm concentration/number was highly heterogeneous and both groups included men ranging from severe oligozoospermia to normozoospermia. Total DFI was significantly lower at the end of the study in homozygous carriers of the p.N680S N versus p.N680S S allele (P = 0.008). Total DFI decreased significantly from baseline to the end of the study (P = 0.021) only in carriers of the p.N680S homozygous N polymorphism, and this decrease involved the sperm population containing vital sperm (i.e. brighter sperm) (P = 0.008). The dimmer sperm DFI fraction, including only nonvital sperm, was significantly larger in p.N680S S homozygous patients than in homozygous N men (P = 0.018). Total DFI was inversely related to total sperm number (P = 0.020) and progressive sperm motility (P = 0.014). When patients were further stratified according to sperm concentration (normoozospermic versus oligozoospermic) or -211G>T polymorphism in the FSHB gene (rs10835638) (homozygous G versus others), the significant improvement of sperm DFI in FSHR p.N680S homozygous N men was independent of sperm concentration and associated with the homozygous FSHB -211G>T homozygous G genotype. The statistical power of the study is 86.9% with alpha error 0.05. This is the first pharmacogenetic study suggesting that FSH treatment induces a significant improvement of total DFI in men carriers of the p.N680S homozygous N FSHR; however, the results need to be confirmed in larger studies using a personalized FSH dosage and treatment duration. The evaluation of sperm DFI as a surrogate marker of sperm quality, and of the FSHR SNP rs6166 (p.N680S), might be useful to predict the response to FSH treatment in men with idiopathic infertility. The study was supported by an unrestricted grant to M.S. and H.M.B. from Merck Serono that provided the drug used in the study. MS received additional grants from Merck Serono and IBSA as well as honoraria from Merck Serono. The remaining authors declare that no conflicts of interest are present. EudraCT number 2010-020240-35.

FSHR gene Thr307Ala and Asn680Ser polymorphisms in infertile men: an association study in North China and meta-analysis

Male infertility is a complex multifactorial and polygenic disease, and genetic factors play an important role in its formation and development. Recently, the association between follicle stimulating hormone receptor (FSHR) gene polymorphisms and male infertility risk has attracted widespread attention due to the unique biological functions of FSH. The aim of this study was to further explore the associations between the Thr307Ala and Asn680Ser polymorphisms of the FSHR gene and male infertility. A case-control study of 212 infertile and 164 fertile men from North China was performed. FSHR polymorphism genotypes were obtained through direct DNA sequencing. A meta-analysis was also performed. In the single-site association analysis, no significant associations were identified between FSHR Thr307Ala and Asn680Ser polymorphisms and male infertility (P > 0.05). However, we found that the combined genotypic frequency of Thr/Ala + Asn/Asn was higher in infertile patients than in controls (6.6 vs 1.8%; odds ratio (OR) = 3.795; 95% confidence interval (CI): 1.072-13.434, P = 0.027). In the meta-analysis, there was also no evidence of FSHR polymorphism (rs 6165 and rs 6168) association with male infertility (P > 0.05). However, we found that the combined genotypes Thr/Thr + Asn/Asn had an increased risk of male infertility (OR = 1.238; 95%CI: 1.001-1.537, P = 0.049). Our studies further confirmed reports that there were no significant associations between the FSHR Thr307Ala and Asn680Ser polymorphisms and male infertility risk. However, a combined FSHR genotype showed significant association with male infertility.

FSHR genotype affects estrogen levels but not pregnancy rates in Luxi cattle subjected to embryo transfer

Follicle-stimulating hormone receptor (FSHR), which mediates the functioning of FSH, plays a central role in reproduction. We investigated bovine FSHR gene polymorphisms and analyzed their relationships with pregnancy rates after embryo transfer and with hormone concentrations on the day of embryo transfer. One reported SNP of FSHR, G-278A, located in the 5'-upstream region, was analyzed and three genotypes (GG, GA and AA) were detected in 132 Luxi cattle recipients. Statistical analysis revealed that recipients with the GG genotype had significantly higher estrogen levels on the day of embryo transfer than did GA and AA genotypes. There were no significant differences in pregnancy rates among genotypes, after embryo transfer. We conclude that variation at these loci of the FSHR gene has no significant effect on pregnancy rates in Luxi cattle.

FSH receptor genotype does not predict metaphase-II oocyte output or fertilization rates in ICSI patients

The objective of this study was to assess the role of the variant p.Asn680Ser in the FSH receptor gene (FSHR) in determining oocyte maturity. It also assessed the relationship between this FSHR variant with metaphase-II oocyte output rate (MOR) and the fertilization rate. This was a prospective observational study based at a tertiary referral centre for reproductive medicine. Women (n=212) undergoing their first cycle of ovarian stimulation for IVF with intracytoplasmic sperm injection (ICSI) were included in the study. Baseline pelvic ultrasound and blood tests were taken on day 2 or 3 of the cycle for assessment of baseline hormones and for DNA extraction. Genotypes for FSHR p.Asn680Ser was determined using TaqMan allelic discrimination assay. The outcome measures were the total dose of exogenous gonadotrophins used, antral follicle count (AFC), number of mature (metaphase-II) oocytes retrieved, MOR and fertilization rate. No statistically significant differences were found between the number of mature oocytes retrieved, MOR or fertilization rates among the patients with different p.Asn680Ser FSHR genotypes. No significant difference was noted in the clinical pregnancy rates per transfer. There is no evidence that the p.Asn680Ser FSHR genotype predicts oocyte maturity.IVF is a complex treatment for infertility requiring costly drugs and carrying significant risk of complications. The procedure aims to stimulate the ovaries to produce more eggs, and conventional methods include a combination of hormones to induce follicle growth, from which eggs are collected. Before beginning the first IVF cycle, knowledge of individual ovarian activity may help predict the response to stimulation and allow clinicians to tailor treatments. Over the last 50years, it has become evident that studying the genetic mechanisms behind variations in drug response in can help control and minimise adverse effects of drugs and their related actions. Hypothetically, a genetic test looking at the variations in the FSH receptor gene may help identify the dosage required and can predict the quality of egg and embryo. One of these variants (at position 680 of exon 10) has been linked to differences in FSH action during ovarian stimulation. None of the studies to date have looked at the role of these genetic markers in the prediction of the quality of eggs and fertilization. We undertook a large prospective study and did not see a relationship between a FSH receptor gene variation and the dosage of gonadotrophins used, quality of eggs or fertilization rates. We surmise that the outcome of ovarian stimulation could be dependent on several factors – both genetic and non-genetic – in a complex multi-factorial model. In the future, genome-wide screening strategies may allow the discovery of further genetic variants associated with ovarian response to stimulation.

Association of rs6166 polymorphism with FSH receptor transcript variants and steroid production in human granulosa cell cultures

Follicle stimulating hormone receptor (FSHR) genetic variation at position 2039 A > G (rs6166, p.Asn680Ser) was repetitively shown to correspond to the measures of ovarian sensitivity and the outcomes of gonadotropin stimulation. However, to date, there has been no study revealing the mechanisms behind the associations observed. The aim of the present research was to investigate the relationship between rs6166 and mRNA expression of FSHR-dependent genes such as LHCGR, CYP19A1, and FSHR itself with particular reference to the FSHR transcript variants (deletions of exon 2, 6, and 9 and insertion of a novel exon between exons 8 and 9) in the cell culture model. Steroid production and its dependency on FSHR genotype were also assessed. A total of 22 normoovulatory patients undergoing IVF treatment were recruited. Granulosa cells were obtained by ovary puncture and cultured for 7 days to regain responsiveness to FSH in a gonadotropin-free medium. Stimulation was carried out for 24 hours in a serum-depleted environment using 0.5 UI/l rhFSH. Gene expression was assessed by real-time PCR and genotype was determined by allele-specific PCR. The distribution of p.Asn680Ser genotypes was as follows: 10 homozygotes Asn/Asn, 8 heterozygotes Asn/Ser, and 4 homozygotes Ser/Ser. Expression of total FSHR in all samples studied increased by a mean factor of 1.9 (95% CI: 0.39–11.53, p < 0.001) upon stimulation. All of the analyzed FSHR transcript variants were detectable in non-stimulated and stimulated cells. The only distinct transcript that followed up-regulation was deletion of exon 2. Homozygotes Asn/Asn tended to have higher rhFSH-induced expression of FSHR as compared to the carriers of Ser/Ser genotype. Relative expression of LHCGR and CYP19A1 although up-regulated showed no significant difference with respect to the FSHR genotype. Variable modulation of FSHR expression by its own ligand is likely to explain different clinical behavior of patients with FSHR genetic variants. The putative contribution of rs6166 requires further investigation.

FSH receptor polymorphism predicts outcome of ovulation induction in WHO-II anovulatory subfertility

The classical treatment algorithm in normogonadotropic (WHO-II) anovulatory infertility constitutes ovulation induction using clomiphene citrate as first-line and exogenous gonadotrophins (exFSH) as second-line therapy. Applying the 2003 Rotterdam criteria about 91% of WHO-II anovulatory patients are diagnosed with polycystic ovary syndrome (PCOS). A polymorphic variant of the FSH receptor (rs6166, Asn680Ser) has been associated with changes in the phenotype of women with PCOS i.e., higher basal FSH and LH levels and higher Testosterone levels. The present study aims to assess the influence of this polymorphic variant of the FSH receptor on the outcome of ovulation induction using CC and exFSH. The study was designed as a prospective longitudinal study (academic fertility unit) among 240 consecutive women with WHO-II anovulatory infertility. Results were replicated in an independent retrospective cohort of 185 PCOS patients (Rotterdam criteria). Patients were treated with Clomiphene, followed by second-line treatment with exFSH if necessary. Genotyping was performed in 159 of 240 (first cohort) and 185 (second cohort) women. Main outcome parameters were ovulation rate and ongoing pregnancy rate. Genotyped patients were similar to ungenotyped patients regarding clinical characteristics and outcome of ovulation induction. FSH receptor polymorphism (rs6166) was associated with clomiphene resistant anovulation. A pooled analysis of both cohorts showed 89% higher chance of anovulation (OR 1.9 [95%CI 1.1-3.3]) in homozygous carriers of the FSH receptor variant (Ser680). The pooled analysis did not show an association of ongoing pregnancy rates with FSH receptor genotype. FSH receptor polymorphism is associated with anovulation during treatment with clomiphene citrate. Therefore, FSH receptor genotyping may find its way into clinical practice in order to devise treatment algorithms that are better tailored to the individual patient.

Evaluating the role of the FSH receptor gene Thr307‐Ala and Asn680‐Ser polymorphisms in male infertility and their association with semen quality and reproductive hormones

To determine whether Thr(307)-Asn(680) and Ala(307)-Ser(680) polymorphisms of the follicle-stimulating hormone receptor (FSH-R) gene are associated with male infertility, semen quality, and reproductive hormones. The FSH-R polymorphisms at codons 680 and 307 were analysed by restriction-fragment-length polymorphism (RFLP) in 172 infertile men and in an equal number of age-matched healthy fertile men. Genotyping of the FSH-R gene was performed using the polymerase chain reaction RFLP technique. All of the participants underwent semen analysis, and reproductive hormones were also measured. Allelic frequencies were 29.7% serine (Ser) and 70.3% asparagine (Asn) for fertile men (the control group), and 33.1% Ser and 66.9% Asn for infertile men (P > 0.05). The FSH-R genotype at position 680 was 49.4% (Asn/Asn), 41.9% (Asn/Ser), and 8.7% (Ser/Ser) in the control group and 40.1% (Asn/Asn), 46.5% (Asn/Ser), and 13.4% (Ser/Ser) in infertile men, respectively (P > 0.05, chi-squared test). Allelic frequencies were 33.1% alanine (Ala) and 66.9% threonine (Thr) for the control group, and 37.8% Ala and 62.2% Thr for the infertile men. The frequencies of genotypes at position 307 were 45.5% Thr/Thr, 43% Thr/Ala, and 11.6% Ala/Ala for the control group and 36.1% Thr/Thr, 52.3% Thr/Ala, and 11.6% Ala/Ala for infertile men. No significant association between codon 680 and codon 307 genotypes and infertility was observed (P = 0.076 and P = 0.073, respectively). The odds ratio (OR) values indicated that individuals with the Thr/Thr + Asn/Ser combined genotypes had a > 50% decreased risk for developing infertility (OR = 0.44; 95% confidence interval [CI]: 0.22-0.77; P = 0.006). The patients with heterozygous Thr/Ala + Asn/Ser combined genotype were 2.65 times more susceptible to infertility than the control group (OR = 2.65; 95% CI: 1.74-3.82; P = 0.0053). The FSH-R codon 680 and codon 307 genotypes did not result in different serum FSH levels either in men with normal spermatogenesis (the control group) or in men with oligoasthenoteratozoospermia (infertile men). We did not observe any significant association of FSH-R genotype frequencies with any of the sperm characteristics analysed in either group. No significant correlation between serum FSH levels and semen characteristics, or fertility status and FSH-R gene polymorphisms was found. The combination of heterozygous Thr/Ala + Asn/Ser genotypes increases the risk for male infertility.