FSH Receptor Gene Research Articles

Novel method for mass producing genetically sterile fish from surrogate broodstock via spermatogonial transplantation†.

A stable system for producing sterile domesticated fish is required to prevent genetic contamination to native populations caused by aquaculture escapees. The objective of this study was to develop a system to mass produce stock for aquaculture that is genetically sterile by surrogate broodstock via spermatogonial transplantation (SGTP). We previously discovered that female medaka carrying mutations on the follicle-stimulating hormone receptor (fshr) gene become sterile. In this study, we demonstrated that sterile hybrid recipient females that received spermatogonia isolated from sex-reversed XX males (fshr (-/-)) recovered their fertility and produced only donor-derived fshr (-) X eggs. Natural mating between these females and fshr (-/-) sex-reversed XX males successfully produced large numbers of sterile fshr (-/-) female offspring. In conclusion, we established a new strategy for efficient mass production of sterile fish. This system can be applied to any aquaculture species for which SGTP and methods for producing sterile recipients can be established.

Production and validation of a polyclonal serum against bovine FSH receptor

In ovarian granulosa cells, follicle-stimulating hormone (FSH) regulates the proliferation and differentiation events required for follicular growth and oocyte maturation. FSH actions are mediated exclusively through the FSH receptor (FSHR). In cattle, the FSHR gene expression pattern during folliculogenesis and the implications of this receptor in reproductive disorders have been extensively studied. However, the limited availability of specific antibodies against bovine FSHR has restricted FSHR protein analysis. In the present study, we developed an anti-FSHR polyclonal serum by using a 14-kDa peptide conjugated to maltose binding protein. The antiserum obtained was characterized by western blot of protein extracts from bovine follicles, BGC-1 cells and primary cultures of granulosa cells stimulated with testosterone. Also, the blocking effect of serum on estradiol secretion and cell viability after gonadotropin stimulus was characterized in a functional in vitro assay. A 76-kDa protein, consistent with the predicted molecular size of full-length FSHR, was detected in ovarian tissue. Besides, two immunoreactive bands of 60-kDa and 30-kDa (only in cultured cells) were detected. These bands would be related to some of the isoforms of the receptor. Therefore, immunohistochemical assays allowed detecting FSHR in the cytoplasm of granulosa cells and an increase in its expression as follicles progressed from primordial to large preantral follicles. These results suggest that the anti-FSHR serum here developed has good reactivity and specificity against the native FSHR. Therefore, this antiserum may serve as a valuable tool for future studies of the biological function of FSHR in physiological conditions as well as of the molecular mechanism and functional involvement of FSHR in reproductive disorders.

Gene expression of follicle-stimulating hormone receptors in granulosa cells in poor ovarian responders

Women with poor ovarian response do not produce adequate follicles and express low estradiol levels after standard ovarian stimulation protocols; therefore, their chances of becoming pregnant are reduced. The follicle-stimulating hormone (FSH) binds to specific follicle-stimulating hormone receptors (FSHRs) in the ovary, thereby activating adenylyl cyclase/cyclic adenosine monophosphate, a gene expression regulator. The present study aimed to evaluate whether the FSHR levels in poor ovarian responders differed from those in normal ovarian responders. This cross-sectional study was conducted in the Cipto Mangunkusomo’s Hospital. The study cohort included women with poor ovarian responses (n = 17) and control women with normal ovarian responses (n = 17) who underwent IVF procedures. The FSHR levels in ovarian tissues were measured using qPCR and then compared between the two groups. No statistically significant differences were found in the FSHR levels between the groups. A polymorphism mutation may be present in the FSHR gene of poor ovarian responders.

The impact of FSH receptor polymorphism on time-to-pregnancy: a cross-sectional single-centre study

BackgroundSingle nucleotide polymorphism of the follicle-stimulating hormone (FSH) receptor (FSHR) is an important marker of ovarian function. However, its role in female fecundity remains debatable. The aim of the study to assess the relationship of FSHR polymorphism of Serine/Serine, Asparagine/Asparagine and Asparagine/Serine variants directly against the time-to-pregnancy (TTP) in women.MethodsData were collected from 291 consecutive selected post-partum Caucasians using this criterion: ethnicity, age between 21 and 34-year-old new mothers and, 0–3 days after delivery of newborns in the Klaipeda University Hospital, Lithuania. Questionnaires on factors associated with conception were given to patients, and blood samples were collected for genomic DNA extractions as well as for analysis of follicle-stimulating hormone receptor gene polymorphism. Odds ratios (OR) and 95% confidence intervals (CI) for time-to-pregnancy were estimated by multivariate logistic regression. Women with unplanned pregnancies and those who received assisted reproductive technologies were not included in the study.ResultsAfter adjustment for other possible factors, increased risk for time-to-pregnancy of 12 or more months was associated with: Serine/Serine polymorphism variant (OR = 1.38, 95% CI 1.56–2.71, p = 0.007), age of 30 or more years (OR = 1.95, 95% CI 1.25–2.71, p = 0.015), gynaecological diseases in the past (OR = 2.21, 95% CI 1.12–5.74, p = 0.027), prior contraception use (OR = 1.87, 95% CI 1.14–3.64, p = 0.016), and fertility problems in the past (OR = 1.57, 95% CI 1.16–4.76, p = 0.019).ConclusionThe results suggest a possible relationship of FSH receptor gene Serine/Serine variant for the lower possibility of conception during the first 12 months of planned conception.

Molecular analysis of FSH receptor gene in Iraqi women with PCOS syndrome

Abstract Polycystic Ovarian Syndrome (PCOS) is a common endocrine disorder in women during their reproductive age. This study was design to establish the relationship between this syndrome and follicle stimulating hormone receptor defect by determination of lethal single nucleotide polymorphism that may play a vital role in this syndrome. A total number of 500 women attending Kamal Al- Samarrai Hospital diagnosed with PCOS were selected and divided according to their age into group one which includes (20–30) years old women, group two included (31–40) years old women, and group three which included (41–50) years old women. Fertility hormones (FSH, LH, and testosterone) were tested for all groups. Results showed that LH increased significantly in groups three with low FSH, whereas testosterone increased significantly in age group two. Molecular analysis of whole FSHR gene amplified using specific primers showed the presence two SNPs rs6166, and rs6165 which are associated with drug response and 9 lethal missense mutation that caused sever effect on FSHR, that probably render this receptor more sensitive to FSH without the possibility of feed back inhibition.

The research of the progesterone and estrogen secretion level and the FSHR, LHR gene quantitative in granular cells cultured in vitro in different time of laying hens

To research the hormone secretion levels of progesterone and estrogen and the gene expression levels of two go-nadotropin receptors follicle stimulating hormone receptor (FSHR) and luteinizing hormone receptor (LHR) in granular cells of laying hen, and the effect of culture time on the levels of hormone secretion and expression of related receptor gene in granulosa cells was inferred. The experiment using the method of cells culture in vitro, the granular cells supernatants of hens were collected at 0 h, 24 h, 48 h, 72 h, 96 h, the progesterone and estrogen concentrations in cell supernatants were determined by ELISA kits, and detected the expression of FSHR and LHR gene in granular cells by real-time fluorescent quantitative PCR. The results showed that the progesterone and estrogen secretion reduced in the early culture of 0 h~48 h(P < 0.05), with the culture time increases to 72 h, the secretion of two hormones began to in-crease, and reaching the level of the initial level of culture. When the cells were cultured to 96 h, the rogesterone and estrogen secretion was reduced again. The lower levels of FSHR and LHR mRNA expression in granular cells appeared with the increase of culture time, compared with the group of cell culture to 0 h, the mRNA expression levels of each groups reduced obviously(P < 0.05). The amount of progesterone and estrogen in the cultured follicular granulosa cells decreased with the increase of in vitro culture time, and then increased. This might be related to the growth state of cells cultured in vitro. But on the whole, with the extension of the training time, the secretion of proges-terone and estrogen in the cells decreased. This may be related to the decreased expression of the FSHR and LHR genes in the two go-nadotropin receptors.

Association of estrogen, progesterone and follicle stimulating hormone receptor polymorphisms with in vitro fertilization outcomes

ABSTRACTDespite the advances in in vitro fertilization (IVF), the implantation success rate for infertile women remains approximately only 15%. In this study, we sought to determine whether implantation failure after repeated IVF treatments is influenced by the presence of common variants in estrogen α, progesterone and follicle stimulating hormone receptor genes. The study population included three groups of women: group 1 were 50 women who had the transfer of ≥3 high-quality embryos during the IVF procedure without ever having had a clinical pregnancy; group 2 were 50 women who achieved a clinical pregnancy after ≤3 high-quality embryos transfers and group 3 were 50 control subjects who achieved a clinical pregnancy without any fertility therapy that resulted in a one live-born infant. Genotype analysis was performed using polymerase chain reaction and Sanger sequencing for rs6165, rs6166, rs2234693, rs9340799. While progesterone receptor single nucleotide polymorphism (SNP) was genotyped based on the amplicon size, the repeats for the ESR1 TA-repeat polymorphism were calculated based on the fragment length. A higher frequency of the heterozygote AG genotype was observed in the infertile groups when compared to controls. Significantly, an allele combination of T of rs2234693, A of rs9340799; S of ESR1 (TA), A of rs6166, G of rs6165 and del of PROGINS had a higher frequency in women who had a successful IVF outcome compared to women who had an unsuccessful IVF outcome, indicating a possible protective combined genotype that could reduce a negative outcome during IVF. This study has demonstrated that combining several candidate genes is needed to assess which may play a role in fertility.Abbreviations: CI: confidence interval; COH: controlled ovarian hyperstimulation; DNA: deoxyribonucleic acid; ESR: estrogen receptors; FSH: follicle stimulating hormones; FSHR: FSH receptor; IVF: in vitro fertilization; PGR: progesterone receptors; SNP: single nucleotide polymorphism

Impact of Lactational Exposure to Polychlorinated Biphenyl Causes Epigenetic Modification and Impairs Sertoli Cells Functional Regulators in F1 Progeny

Polychlorinated biphenyl (PCB) is an endocrine-disrupting chemical. Sertoli cells (SCs) provide physical and nutritional support for developing germ cells. Dysfunction in SCs has adverse effects on spermatogenesis. Previously, we found that the lactational exposure of PCBs (1, 2, and 5 mg/kg birth weight/day, orally from postnatal days 1 to 20) decreased the follicle-stimulating hormone receptor (FSHR) and androgen receptor (AR) expression in SCs of F1 progeny. Transcription factors initiate and regulate the transcription of genes. DNA methylation plays an important role in epigenetic gene regulation. Hence, this study was aimed to identify the level of transcription factors regulating FSHR, AR gene expression, and DNA methylation in the promoter of these genes in SCs of both F1 prepuberal and puberal offspring. DNA methylation in the promoter of FSHR and AR genes was examined by sodium bisulfite conversion technique. The protein levels of transcription factors (steroidogenic factor 1 [SF1], upstream stimulatory factors 1 and 2, c-fos, c-jun, and CREB-binding protein) and enzymes DNA methyltransferases (Dnmt1, Dnmt3ab, Dnmt3l, and histone deacetylase 1 [HDAC1]) were analyzed by Western blotting. The transcription factors that regulate the FSHR and AR gene in SCs were decreased in both the PCB-exposed F1 progeny. Methylation was observed in the promoter of FSHR, AR, and SF1. The protein levels of Dnmt1, Dnmt3ab, Dnmt3l, and HDAC1 were increased in the PCBs-treated groups. Subsequently, it leads to transcriptional repression of the genes in SCs. Our finding suggests that PCBs caused epigenetic change in SCs, thereby it impaired SCs function in F1 progeny.

Polymorphism FSHR (-29G/A) as a genetic agent together with ESRI (XbaIG/A) in women with poor response to controlled ovarian hyperstimulation.

One of the major problems in IVF is the poor response of the ovary to gonadotropins. ESRI and FSHR are two effective genes on controlled ovarian hyperstimulation (COH). Evaluating the correlation of alleles and genotypes of polymorphism (-29G/A) located in the FSH receptor gene and polymorphism (XbaI G/A) located in Estrogen receptor genes with the ovary's response would help to anticipate the results of ovulation in IVF cycles. In the present study, two hundred (200) blood samples were taken from infertile women aged 20 to 39 who were under IVF therapy. After DNA extraction from the samples, real-time PCR was performed using a specific probe-primer. Statistical analysis revealed that the frequency of alleles and genotypes of polymorphisms (-29G/A) and (XbaI A/G) in women with normal to poor response did not have significant correlation.

The Impact of FSHR Gene Polymorphisms Ala307Thr and Asn680Ser in the Endometriosis Development

Endometriosis is an estrogen-dependent inflammatory disease that affects a large number of women in reproductive age. Follicle-stimulating hormone (FSH) plays a role in steroidogenesis and acts through a transmembrane glycoprotein, FSH receptor (FSHR). Polymorphisms in FSHR gene were previously associated with variability in FSH serum level and reproductive outcomes, but its relation with endometriosis has not been clarified and demonstrated conflicting results, ranging from strong links to no association to endometriosis. Inspired by these findings, we aimed to investigate the influence of FSHR Ala307Thr and Asn680Ser polymorphisms in the risk of endometriosis development and/or progression and the status of fertility in 352 women with endometriosis and 510 fertile controls. Single-marker analysis revealed no significant difference for both Ala307Thr and Asn680Ser polymorphisms between overall endometriosis and control group. However, when the endometriosis group was subdivided according to fertility status and disease stage, a positive association was found between 680Ser/Ser or GG genotype of the Asn680Ser polymorphism and fertile women with endometriosis (p = 0.004). Combined alleles of FSHR polymorphisms revealed that "GG/307Ala680Ser" was more frequently found in fertile women with endometriosis (haplotype frequency of 45.4% in fertile women with endometriosis and 38.3% in controls, p = 0.041). The combined alleles of FSHR polymorphisms disclosed that "GG/307Ala680Ser" was more frequently found in fertile women with endometriosis (haplotype frequency of 45.4% in fertile women with endometriosis and 38.3% in controls, p = 0.049), while "GA/307Ala680Asn" haplotype was less frequently found in endometriosis group (haplotype frequency of 6.5% in cases and 11.9% in controls, p = < 0.001), regardless of fertility status and stage of the disease. The findings suggest that 680Ser-Ser/GG genotype and "GG/307Ala680Ser" haplotype increase the risk of endometriosis in fertile women, while "GA/307Ala680Asn" haplotype decreases the risk of endometriosis development and progression.

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.

Vitrification and in vitro culture had no adverse effect on the follicular development and gene expression of stimulated human ovarian tissue.

The study assesses the effect of the vitrification procedure on the integrity, morphology, follicular development and gene expression of stimulated human ovarian tissue after warming and two weeks of in vitro culture. Ovarian specimens were divided into non-vitrified and vitrified groups and were cultured for two weeks. Morphological analysis and immunohistochemistry were performed. The 17-β estradiol and anti-Müllerian hormone levels in collected media were assessed. Gene expression was analyzed using real-time reverse transcription polymerase chain reaction. The morphology and immunohistochemistry of bcl-2-like protein 4 and B-cell lymphoma 2 of human stimulated ovarian tissue were similar in both groups. There was no significant difference in the percentage of normal follicles between the groups before and after in vitro culture. In spite of an increase in the percentage of growing follicles in cultured tissues compared to the non-cultured groups, the rate of normal follicles was significantly decreased in both cultured groups (P < 0.05). Gene expression was no different in vitrified tissues compared to the control; however, the expression of growth differentiation factor 9 and follicle stimulating hormone receptor genes were increased and factor in germ line alpha and kit ligand genes were decreased during in vitro culture (P < 0.05). In the two cultured groups, the level of 17-β estradiol was increased (P < 0.05), but the anti-Müllerian hormone concentration was not statistically altered. These results showed that the integrity of stimulated human ovarian tissue after vitrification/warming was well preserved; however, the in vitro culture condition needs improvement.

Leutinizing hormone/choriogonadotropin receptor and follicle stimulating hormone receptor gene variants and risk of recurrent pregnancy loss: A case control study

We investigated the association of luteinizing hormone/chorionic gonadotropin receptor (LHCGR) and follicle stimulating hormone receptor (FSHR) polymorphisms with recurrent pregnancy loss (RPL). This was a retrospective case-control study involving 308 women with RPL and 310 control women. FSHR and LHCGR genotyping was done by allelic exclusion method. Lower minor allele frequency (MAF) of LHCGR rs7371084, and rs4597581, and FSHR rs6166, and higher MAF of rs1007541, and rs2055571 FSHR variants were seen in RPL cases. Significantly lower frequencies of heterozygous LHCGR rs2293275, rs7371084, and rs4597581, and homozygous FSHR rs6166, and higher frequency of homozygous FSHR rs2055571 carriers were seen in RPL cases, after controlling for key covariates. LHCGR rs7371084 and rs4597581 were negatively associated with RPL in non-obese subjects. On the other hand, FSHR rs1007541 and rs2055571 were positively, while FSHR rs6166 was negatively associated with RPL in obese subjects. Limited linkage disequilibrium was seen only among FSHR variants; higher frequency of GTAAA and AAGGA, and lower frequency of GTGGG FSHR haplotypes was seen in RPL cases; the frequencies of remaining FSHR haplotypes, and all LHCGR haplotypes were similar between cases and controls. This is the first study to demonstrate association of novel LHCGR (rs7371084 and rs4597581) and FSHR (rs6166, rs1007541 and rs2055571) SNPs with RPL, particularly according to obesity.

Expression of follicle-stimulating hormone receptor (FSHR), protein kinase B-2 (AKT2) and adapter protein with PH domain, PTB domain, and leucine zipper (APPL1) in pig ovaries.

Follicle-stimulating hormone (FSH) regulates oogenesis and spermatogenesis by binding to its receptor (FSHR) on target cells in the ovary and testis, respectively. The signaling cascades activated after ligand binding are extremely complex and have been shown to include protein kinase A and phosphatidylinositol 3-kinase/protein kinase. The adapter protein APPL1 (adapter protein with PH domain, PTB domain, and leucine zipper), which is an assortment of other signaling proteins, was previously identified to interact with the FSH receptor (FSHR) and the protein kinase B (AKT) pathway. APPL1 plays an important role in promoting cell survival within the preovulatory follicle granulosa layer. Here, we aimed to evaluate the FSHR, AKT2, and APPL1 gene and protein expression levels in the ovaries of different prolific porcine breeds (Wannan Black [WB] and Large White [LW] pigs) using immunohistochemistry and qRT-PCR, respectively. Our results showed that FSHR, AKT2, and APPL1 mRNA levels were significantly higher (P < 0.05) in the ovaries of WB pigs than in the ovaries of LW pigs. Additionally, the FSHR, AKT2, and APPL1 proteins were mainly found distributed in the granulosa cells and oocytes. This study showed that high levels of FSHR, AKT2, and APPL1 were expressed in the ovaries of high prolific breed pigs.

A novel homozygous mutation in the FSHR gene is causative for primary ovarian insufficiency

To identify the potential FSHR mutation in a Chinese woman with primary ovarian insufficiency (POI). Genetic and functional studies. University-based reproductive medicine center. A POI patient, her family members, and another 192 control women with regular menstruation. Ovarian biopsy was performed in the patient. Sanger sequencing was carried out for the patient, her sister, and parents. The novel variant identified was further confirmed with the use of control subjects. Sanger sequencing and genotype analysis to identify the potential variant of the FSHR gene; hematoxylin and eosin staining of the ovarian section to observe the follicular development; Western blotting and immunofluorescence to detect FSH receptor (FSHR) expression; and cyclic adenosine monophosphate (cAMP) assay to monitor FSH-induced signaling. Histologic examination of the ovaries in the patient revealed follicular development up to the early antral stage. Mutational screening and genotype analysis of the FSHR gene identified a novel homozygous mutation c.175C>T (p.R59X) in exon 2, which was inherited in the autosomal recessive mode from her heterozygous parents but was absent in her sister and the 192 control women. Functional studies demonstrated that invitro the nonsense mutation caused the loss of full-length FSHR expression and that p.R59X mutant showed no response to FSH stimulation in the cAMP level. The mutation p.R59X in FSHR is causative for POI by means of arresting folliculogenesis.

Characterization of polymorphism in the FSH receptor gene and its impact on some reproductive indices in dairy cows

Follicle stimulating hormone (FSH) is released from the anterior pituitary gland and has an important role in female fertility. As FSH is a glycoprotein polypeptide hormone which cannot pass through the cell membrane, its influence on target cells must be mediated by the FSH receptor (FSHR). Accordingly, any kind of mutation in FSHR can affect reproduction in dairy cows. In this study, the polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) technique was used for recognition of a point mutation (A/G: position −278) located in the FSHR gene in Iranian dairy cows. The association was evaluated of this mutation with reproductive performance. Blood samples were collected from 79 cows in a dairy farm in Iran and genotyped based on this single nucleotide polymorphism (SNP). The 5′-flanking regions of FSHR gene were successfully amplified and produced a fragment of 211bp in all cases. Three different patterns were, however, produced following restriction digestion with FaqI enzyme. The molecular results showed the existence of three different genotypes of AA, AG and GG among examined cows. In this study percentages of genotypes were 51.9%, 43.2% and 4.9% for AA, AG and GG genotypes, respectively. Allele frequencies were 73.5% and 26.5% for A and G, respectively. Results indicate that cows lacking allele G had desirable fertility in which a greater percentage (53.7%) of cows lacking Allele G (AA) had services per conception (SPC) of <2 in the previous lactation; while a lesser percentage of cows with Allele G (28.9%) had SPC of <2 (P<0.05). There was no difference in the days non-pregnant (DNP) and calving to first service interval among cows with these genotypes (P>0.05). Calving to first service interval was 69.9 ±12.3 in cows with Allele G and 74.73±13.9 in cows without Allele G (P>0.05). Percentage of cows with repeat breeder syndrome (SPC >3) was also 15.6% and 27.6% in cows without Allele G and with Allele G, respectively, but these values were not different (P>0.05). It can be concluded that the A to G mutation within the upstream region of FSHR gene (position −278) may affect some reproductive variables in Holstein dairy cows.

Association of Exon 10A and 10B inactivating mutation of follicle stimulating hormone receptor gene (FSHR) and Polycystic Ovarian Syndrome in Vellore cohort

Polycystic ovarian syndrome is the most common heterogenous endocrine disorder in women. Follicle stimulating hormone receptor is associated with normal development as well as maturation of follicles and triggers estrogen production in granulosa cells of the ovary. Inactivating mutation in FSHR gene correlated with reduction of ovarian function in women is due to damage to receptor function. This study aims to investigate whether inactivating mutations, in follicle stimulating hormone receptor gene is related to polycystic ovarian morphology in women with PCOS. Genomic DNA isolated from 15 subjects from Sandhya Hospital, Vellore (10 patients with PCOS and 5 healthy controls) was taken for this study. Patient data included a clinical report, hormonal levels, and ovarian morphological details. DNA isolation was followed by DNA amplification by polymerase chain reaction using Exon 10 A and Exon 10 B primers. The PCR-RFLP analysis was performed using Dde1 restriction enzyme. Here we discuss inactivating mutation found in Exon 10 of FSHR gene in patients with PCOS.The absence of inactivating mutation was observed through PCR-RFLP study on Exon 10A and Exon 10B.

Effects of silencing INHα gene by RNAi on the mRNA expressions of StAR, FST, INHβB, and FSHR genes in cultured sheep granulosa cells

The aim of the study was to investigate the effects of silencing the inhibin α-subunit (INHα) gene by RNAi on the mRNA levels of steroidogenic acute regulatory protein (StAR), follistatin (FST), inhibin β-subunit (INHβB), and FSH receptor (FSHR) in cultured sheep granulosa cells (GCs). The antral follicles, 3–7mm in diameter, were dissected from the ovaries of 15 Thin-tailed Han sheep (1-1.5 years old), and then GCs were isolated from antral follicles. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to analyze the mRNA levels of StAR, FST, INHβB, and FSHR in INHα-specific small RNA interference (siRNA) − treated GCs. Knockdown of INHα mRNA by siRNA decreased the mRNA expressions of StAR and FST by 76% and 23%, respectively (P<0.05), and increased the mRNA expressions of FSHR and INHβB by 81% and 50% in cultured sheep GCs, respectively (P<0.05). These results indicated that silencing INHα gene by RNAi influenced the mRNA expressions of StAR, FST, INHβB and FSHR in cultured sheep GCs.

Long Noncoding RNAs: Potential Regulators Involved in the Pathogenesis of Polycystic Ovary Syndrome.

Polycystic ovary syndrome (PCOS) is the most common cause of anovulatory infertility in women of reproductive age, and its etiology remains poorly understood. Altered activities of long noncoding RNAs (lncRNAs) have been associated with human diseases and development. However, the roles of lncRNAs are unknown in reproductive medicine. We investigated the potential role of lncRNAs in the pathogenesis of PCOS, using human granulosa cells (GCs) and the KGN cell line. We used microarrays to compare lncRNA expression profiles in GCs from seven patients with PCOS and seven matched women. GC samples were collected during 2014 to 2016 from infertile women in Guangzhou, China. Quantitative real-time polymerase chain reaction was used to measure levels of the lncRNA HCG26 in GCs from 53 patients with PCOS and 50 controls. HCG26 was knocked down with locked nucleic acid GapmeRs in KGN cells to examine its role in cell proliferation, aromatase and follicle-stimulating hormone receptor gene expression, and estradiol production. A total of 862 lncRNA transcripts and 998 messenger RNA transcripts were differentially expressed (greater than or equal to twofold change; P < 0.05) in PCOS GCs compared with those of controls. HCG26 levels were upregulated in patients with PCOS and were associated with antral follicle count. HCG26 knockdown in KGN cells inhibited cell proliferation and cell-cycle progression and increased aromatase gene expression and estradiol production. Our study reports the lncRNA profiles in GCs from patients who have PCOS and those from healthy women and suggests that dysregulated lncRNAs may play vital roles in GC proliferation and steroidogenesis, providing insights into the pathogenesis of PCOS.

The susceptibility of FSHB -211G\xa0>\xa0T and FSHR G-29A, 919A\xa0>\xa0G, 2039A\xa0>\xa0G polymorphisms to men infertility: an association study and meta-analysis

BackgroundMale infertility is a complex disorder caused by genetic, developmental, endocrine, or environmental factors as well as unknown etiology. Polymorphisms in the follicle stimulating hormone beta subunit (FSHB) (rs10835638, c.-211G > T) and follicle stimulating hormone receptor (FSHR) (rs1394205, c.-29G > A; rs6165, c.919A > G; rs6166, c.2039 A > G) genes might disturb normal spermatogenesis and affect male reproductive ability.MethodsTo further ascertain the aforementioned effects, we conducted a case-control study of 255 infertile men and 340 fertile controls from South China using the Mass ARRAY method, which was analyzed by the t-tests and logistic regression analysis using SPSS for Windows 14.0. In addition, a meta-analysis was performed by combining our results with previous reports using STATA 12.0.ResultsIn the FSHB or FSHR gene single nucleotide polymorphism (SNP) evaluation, no statistically-significant difference was found in the frequency of allelic variants or in genotype distribution between cases and controls. However, a significant association for the comparison of GAA (P: 0.022, OR: 0.63, 95%CI: 0.43–0.94) was seen between the oligozoospermia and controls in haplotype analysis of rs1394205/rs6165/rs6166. In the meta-analysis, rs6165G allele and rs6166 GG genotype were associated with increased risk of the male infertility.ConclusionsThis study suggested that FSHR GAA haplotype would exert protective effects against male sterility, which indicated that the combination of three SNP genotypes of FSHR was predicted to have a much stronger impact than either one alone. Then in the meta-analysis, a significant association was seen between FSHR rs6165, rs6166 polymorphisms and male infertility. In terms of male infertility with multifactorial etiology, further studies with larger sample sizes and different ethnic backgrounds or other risk factors are warranted to clarify the potential role of FSHB and FSHR polymorphisms in the pathogenesis of male infertility.