FSH Receptor Gene Research Articles

Genetic and Biochemical Markers of Ovarian Function and Their Impact on Women Reproductive Status

Endometriosis is a disease very common nowadays affecting 1-2% of the female population, by estrogen-dependent mechanism. The identification of mutations in the gene encoding for the FSH receptor (FSHR) has been reported since 1995. Physiology teaches us that follicle-stimulating hormone (FSH) is a hormone that is vital in the steroidogenesis regulation mechanisms, while FSH receptor (FSHR) activation helps to promote folliculogenesis and estrogensynthesis. Therefore, studies to show if there are any correlations between endometriosis and FSHR are acquired. Genotyping of FSHR gene polymorphisms were performed using PCR - Restriction Fragment Length Polymorphism (PCR-RFLP) analysis. We analysed a total of 78 patients, 44 infertile patients with endometriosis and 34 controls (non-infertile, pregnant patients). The endometriosis group included women with diagnosis of endo-metriosis confirmed by laparoscopy and /or laparotomy and histological evidence of disease with the endometriosis staging according to American Society for Reproductive Medicine (ASRM). Corroborated with the severity of endometriosis, A919G and A2039G tests found that 71.4% of the M (GG) results were associated with primary infertility, not statistically significant (p=0.994) and 42.9% of the total M results had moderate or severe forms of endometriosis (p = 0.185). The genetic involvement in different pathologies such as endometriosis, has yet to be understood, but knowing more about its mechanism, will help physician target the disease at a more profound level.

Follicle-stimulating hormone receptor gene polymorphisms of ovarian reserve markers in Romanian population.

Association between phenotype and follicle-stimulating hormone (FSH) receptor and FSH beta chain genotype was evaluated in women with ovarian dysfunction. FSH receptor gene single nucleotide polymorphisms (SNPs) were analyzed by restricted fragment length polymorphism (RFLP) technique. Three groups were analyzed: two groups formed of poor responders (women with ovarian dysfunctions caused by endometriosis and patients who underwent ovarian stimulation protocols) and a third good responders group (normal-ovulatory women who gave birth to naturally conceived children). A higher average level of basal FSH values were found in mutants in the A919G/Ala307Thr/rs6165 or A2039G/Asn680Ser/rs6166 tests (7.16±1.09; P=0.659). Anti-mullerian hormone (AMH) below 1.2 ng/ml was associated with a higher frequency of mutations: 33.3% A919G/Ala307Thr and A2039G/Asn680Ser (P=0.137) and also in 66.6% FSH receptor less frequent polymorphism (c.-29G>A) rs 1394205 (P=0.522). The age, day 3 FSH, and AMH levels are widely used to investigate female infertility. However, we have not yet found the ideal biomarker to determine the best outcome and treatment plan for our patients. We cconsider that genetic markers will become the future in the personalization of controlled ovarian stimulation treatment in the upcoming period.

Melatonin regulates the expression of Bone Morphogenetic Protein 15 (Bmp-15), Growth Differentiation Factor 9 (Gdf-9) and LH receptor (Lhr) genes in developing follicles of rats

This study investigated the effects of melatonin supplementation on the daily mRNA expression of Bmp-15, Gdf-9, FSH (Fshr) and LH (Lhr) receptor genes and the effect of gonadotropin supplementation on protein expression of melatonin-synthetizing enzymes (ASMT and AANAT) during in vitro maturation (IMV) of rat follicle-enclosed oocytes. We also studied the effects of pinealectomy on the mRNA expression of Bmp-15, Gdf-9, Fshr and Lhr genes in immature oocytes and cumulus cells and on the protein expression and immunolocalization of AANAT and ASMT in immature ovaries. Ovaries were collected from sham-pinealectomized (INTACT) and pinealectomized (PINX) females during the light and dark phases of the 12h:12h light-dark cycle. Melatonin increased the Bmp-15 and Gdf-9 mRNA expression in follicle-enclosed oocytes during dark phase and Lhr mRNA expression during the light phase. The daily mRNA expression of Lhr was negatively correlated with the daily mRNA expression of Bmp-15 and Gdf-9 in follicle-enclosed oocytes supplemented with melatonin. There were no phase differences in oocytes mRNA expression of Bmp-15 and Gdf-9 from PINX females. Pinealectomy did not altered the mRNA expression of Lhr gene in cumulus cells. There were no phase differences in ASMT and AANAT expression in ovaries from INTACT and PINX females and their immunolocalization was seen in most of ovarian cells. AANAT expression in follicle-enclosed oocytes supplemented with LH from INTACT and PINX females was greater during the dark phase. The results suggest that melatonin regulates the daily Bmp-15, Gdf-9 and Lhr mRNA expression during the rat follicle development. There is also an indicative that melatonin plays an important role in the relationship between the daily expression of Lhr and the daily expression of Bmp-15 and Gdf-9 in follicle-enclosed oocytes during oocyte maturation process.

DNA polymorphism of FSHR gene and its association with infertility traits in Egyptian buffaloes

The objectives of this study were to detect genetic polymorphism in exon 10 of Follicle Stimulating Hormone Receptor gene (FSHR) by using RFLP and direct sequence methods, also to investigate their possible association with infertility traits in Egyptian buffaloes. A total number of 50 female Egyptian buffaloes at Mahallet Mousa experimental farms in Kafr El-sheikh province were investigated. Animals were divided into three groups: normal fertile (20 animals), infertile due to anestrum (15 animals) and infertile due to repeat breeding (15 animals). Genomic DNA was extracted from blood samples, then PCR was performed with annealing temperatures of 57oC to amplify 231 bp of FSHR exon 10 gene. The amplicons were digested with Hin1II (NlaIII) restriction enzyme. The results showed that all the investigated buffaloes were monomorphic and genotyped as A/A genotype. Moreover, the monomorphism of buffaloes FSHR exon 10 gene was confirmed by sequencing three samples from each buffaloes’ group. It seems that FSHR gene are highly conserved in the investigated buffalo’s population.

The 2039 A/G FSH receptor gene polymorphism influences glucose metabolism in healthy men

ObjectiveTo assess the role of c. 2039 A/G (p. Asp680Ser) (rs6166) and c. −29 G/A (rs1394205) follicle-stimulating hormone receptor (FSHR) gene single nucleotide polymorphisms (SNPs) in a cohort of healthy men.MethodsOne-hundred twenty-seven healthy men underwent evaluation of the anthropometric parameters, assessment of metabolic and lipid profile, measurement FSH serum levels, and genotyping of both the aforementioned FSHR SNPs. Data grouped according to the FSHR rs6166 or rs1394205 genotypes underwent to statistical analysis.Main resultsThe three groups of men for each FSHR SNP did not differ statistically significantly for body mass index and serum FSH levels. As for FSHR rs6166 SNP, glucose levels were significantly lower in men with the GG genotype compared with those with the AA genotype. Men with AG had lower insulin levels and HOMA index values compared with those carrying the genotype AA (p < 0.05). The GG group showed a negative correlation between serum FSH levels and insulin and between serum FSH levels and HOMA index (p < 0.05). In contrast, men grouped according to the FSHR rs1394205 genotype showed no significant difference in blood glucose, serum insulin levels, and HOMA index. The AG group showed a negative correlation between FSH insulin and between serum FSH levels and HOMA index (p < 0.05).ConclusionsMen with the genotype GG of the FSHR rs6166 SNP have lower blood glucose levels than those with the AA genotype. Their FSH levels inversely correlated with insulin and HOMA index. In contrast, the genotype FSHR rs6166 A/G did not reveal any role of FSH on glucose metabolism in healthy men. The inverse relationship between FSH and insulin or HOMA index in the group with the genotype GG of the FSHR rs6166 SNP suggests a possible cross-talk between FSH and insulin.

Prospects for FSH Treatment of Male Infertility.

Despite the new opportunities provided by assisted reproductive technology (ART), male infertility treatment is far from being optimized. One possibility, based on pathophysiological evidence, is to stimulate spermatogenesis with gonadotropins. We conducted a comprehensive systematic PubMed literature review, up to January 2020, of studies evaluating the genetic basis of follicle-stimulating hormone (FSH) action, the role of FSH in spermatogenesis, and the effects of its administration in male infertility. Manuscripts evaluating the role of genetic polymorphisms and FSH administration in women undergoing ART were considered whenever relevant. FSH treatment has been successfully used in hypogonadotropic hypogonadism, but with questionable results in idiopathic male infertility. A limitation of this approach is that treatment plans for male infertility have been borrowed from hypogonadism, without daring to overstimulate, as is done in women undergoing ART. FSH effectiveness depends not only on its serum levels, but also on individual genetic variants able to determine hormonal levels, activity, and receptor response. Single-nucleotide polymorphisms in the follicle-stimulating hormone subunit beta (FSHB) and follicle-stimulating hormone receptor (FSHR) genes have been described, with some of them affecting testicular volume and sperm output. The FSHR p.N680S and the FSHB -211G>T variants could be genetic markers to predict FSH response. FSH may be helpful to increase sperm production in infertile men, even if the evidence to recommend the use of FSH in this setting is weak. Placebo-controlled clinical trials, considering the FSHB-FSHR haplotype, are needed to define the most effective dosage, the best treatment length, and the criteria to select candidate responder patients.

The effects of follicle-stimulating hormone receptor (FSHR) -29 and Ser680Asn polymorphisms in IVF/ICSI.

Background The aim of this study was to analyze two different polymorphisms, Ser680Asn and -29 (G>A) promoter polymorphism, of the follicle-stimulating hormone receptor (FSHR) gene, individually but also in combination, in a sample of Greek women undergoing in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). Materials and methods One hundred and forty-one women undergoing IVF or ICSI and 94 controls were genotyped by real-time polymerase chain reaction (RT-PCR) for the two FSHR polymorphisms. The association of the alleles with the clinical, biochemical and other parameters concerning the controlled ovarian stimulation (COS) protocol and outcome was investigated, as well as the pregnancy rate. Results The study of each polymorphism individually revealed a positive correlation of the SerSer genotype (Ser680Asn polymorphism) with higher luteinizing hormone (LH) levels on the third day of the menstrual cycle. On the other hand, the A allele for the -29 (G>A) promoter polymorphism correlated with the increased number and quality of cumulus-oocyte complexes (COCs). No differences were detected when the different genotypes of the two polymorphisms were combined - the population study was grouped according to the number of polymorphic alleles they carried (0-4 alleles). Women who presented all polymorphic alleles, AsnAsn/AA, exhibited the lowest LH levels (2.62 ± 0.68 mIU/L), but were rarely detected (n = 2, 1.4% of the studied population). Conclusions The data from this study reflect that the investigation of the combination of polymorphisms, such as FSHR -29 and Ser680Asn, could offer a valuable tool in order to evaluate and anticipate the outcome of the ovulation induction protocols, especially in the group of patients with failed attempts.

Target prediction and validation of microRNAs expressed from FSHR and aromatase genes in human ovarian granulosa cells

MicroRNAs (miRNAs) are known post-transcriptional regulators of various biological processes including ovarian follicle development. We have previously identified miRNAs from human pre-ovulatory ovarian granulosa cells that are expressed from the intronic regions of two key genes in normal follicular development: FSH receptor (FSHR) and CYP19A1, the latter encoding the aromatase enzyme. The present study aims to identify the target genes regulated by these miRNAs: hsa-miR-548ba and hsa-miR-7973, respectively. The miRNAs of interest were transfected into KGN cell line and the gene expression changes were analyzed by Affymetrix microarray. Potential miRNA-regulated genes were further filtered by bioinformatic target prediction algorithms and validated for direct miRNA:mRNA binding by luciferase reporter assay. LIFR, PTEN, NEO1 and SP110 were confirmed as targets for hsa-miR-548ba. Hsa-miR-7973 target genes ADAM19, PXDN and FMNL3 also passed all verification steps. Additionally, the expression pattern of the miRNAs was studied in human primary cumulus granulosa cell culture in relation to the expression of their host genes and FSH stimulation. Based on our findings we propose the involvement of hsa-miR-548ba in the regulation of follicle growth and activation via LIFR and PTEN. Hsa-miR-7973 may be implicated in the modulation of extracellular matrix and cell-cell interactions by regulating the expression of its identified targets.

Single Nucleotide Polymorphism of Follicle Stimulating Hormone Receptor Gene in Iraqi Infertile Men

The study was directed to determine Single Nucleotide Polymorphism (rs6166 A&gt;G) of follicle stimulating hormone receptor gene and their association as a risk factor with male infertility in Iraqi sterile patient. In the present study after seminal and serum analysis 50 patient (25 Azoospermia and 25 oligozoospermia) and 50 healthy fertile control were joined. The samples were collected from the Biotechnology Department/college of science/Baghdad university and Kamal Al-Samarie IVF Hospital, Baghdad, Iraq. through the period of two months from November 2018 to January 2019. The SNP (rs 6166) determination was carried out by using real-time polymerase chain reaction (qPCR) of blood samples. The difference in the mean of genotype showed a significantly different (p&lt;0.05) in infertile patients group likened to corresponding means infertile control group. Survey of follicle stimulating hormone receptor gene SNP genotypes and allele frequencies in Azoospermia and oligozoospermia patient groups with the control group, showed that there was a significant variation in the heterozygous (AG) and homozygous mutant (GG) genotype frequencies in (rs 6166). It concluded that this SNP may have a role in an Azoospermia and oligozoospermia Iraqi patients complaining from idiopathic infertility.

Follicle –Stimulating Hormone Receptor Polymorphisms in Iraqi Women with Primary Amenorrhea

The study was carried to determine Single Nucleotide Polymorphism (rs6165) of Follicle Stimulating Hormone Receptor (FSHR) gene in blood samples of 62 Iraqi women with primary amenorrhea and 40 healthy control females. The research included chromosomal study and serum analysis of 62 patient samples. The samples were collected from Educational laboratories, City of Medicine, Baghdad and Biotechnology Dept. College of Science- Baghdad University through the period from October 2018 to March 2019. The determinations of SNP (rs6165) were carried out by real-time PCR. Results of rs6165 genotyping showed significant variations between PA patients and controls, Inspecting FSHR gene genotypes and allele frequencies in PA patients groups with the control group, revealed that there was significant variation in the heterozygous (AG) and homozygous mutant type(AA) genotype frequencies in (rs6165). the SNP of target gene may have a role in PA patients complaining from idiopathic puberty problems.

Profile of follicle-stimulating hormone and polymorphism of follicle-stimulating hormone receptor in Madrasin cattle with ovarian hypofunction.

Background and Aim:The follicle-stimulating hormone (FSH) gene is an essential regulator of fertility in livestock. This study aims to provide information on the genetic makeup of Madrasin cattle experiencing hypofunction by the FSH profile and FSH receptors (FSHR) polymorphism.Materials and Methods:Blood samples were collected from the Bangkalan regency in Indonesia. DNA was isolated and purified following the extraction protocol of polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism.Results:Our results showed that the FSH gene had a band length of 310 bp and produce two alleles (A and B) with restriction enzymes at 250 bp, 230 bp, and 145 bp. Furthermore, the FSHR gene had a band length of 303 bp and produced two homozygous genotypes: GG at bp 239 and CC at bp 188.Conclusion:Based on these differences, there was no change in allele frequency and genotype between Madura and Madrasin cattle due to crossbreeding with Limousin cattle. Thus, further detailed investigations of Madrasin cattle are required to elucidate the profile of the LH and LHR genes.

THE FREQUENCY OF FOLLICLE-STIMULATING HORMONE RECEPTOR 2039A>G GENE POLYMORPHISM AND THE RISK OF MALE INFERTILITY IN ALBANIAN POPULATION.

SUMMARYThe purpose of this study was to determine the prevalence of allele and genotype variants of the follicle-stimulating hormone receptor (FSHR) gene polymorphic region at position Asn680Ser in the Albanian male population and associate them with the clinical parameters of infertility. The study included 114 infertile men (mean age 35.04±5.85 years) stratified according to the level of spermatogenetic impairment (oligoasthenozoospermia, asthenozoospermia and normospermia) and 112 fertile men (mean age 36.44±7.05 years) with normal semen parameters. Genotyping of the FSHR gene at position 680 was performed by TaqMan genotyping assay. All the participants underwent semen analysis, and serum reproductive hormones (FSH, luteinizing hormone, prolactin and testosterone) were also measured. The FSHR Asn680Ser genotype frequencies were as follows: Asn/Ser 42%, Ser/Ser 33.9% and Asn/Asn 24.1% in the control group, and Asn/Ser 56.1%, Ser/Ser 22.8% and Asn/Asn 21.1% in the whole group of infertile men (χ2-test: P=0.08). There was no statistically significant correlation between serum hormone levels and semen characteristics or between fertility status and FSHR Asn680Ser gene variants in the control group and the group of infertile men. However, adjusted logistic regression analysis (age, body mass index, smoking and alcohol as covariates) revealed increased odds ratio for male infertility among heterozygous Asn/Ser genotype carriers associated with lower values of semen parameters (normal morphology, concentration, total sperm count and motility). In conclusion, our case-control study further confirmed previous reports on no significant association between the FSHR Asn680Ser polymorphisms and male infertility. Nevertheless, the data presented herein indicate that the Asn/Ser genotype may increase the risk of male infertility in Albanian population.

Role of Follicle-stimulating Hormone Receptor Gene (ser680asn) Polymorphism in Patients with Polycystic Ovarian Syndrome

Role of Follicle-stimulating Hormone Receptor Gene (ser680asn) Polymorphism in Patients with Polycystic Ovarian Syndrome

FSH RECEPTOR AND FSH BETA CHAIN POLYMORPHISM INVOLVEMENT IN INFERTILITY AND ENDOMETRIOSIS DISEASE.

The purpose of this study was to evaluate the association between the follicle-stimulating hormone (FSH) receptor (c.-29G>A) and FSH beta chain (c.-280G>T) polymorphisms and endometriosis in Romanian women. We performed the polymorphic analysis of the FSH receptor gene and FSH beta chain in 44 patients with endometriosis and 34 controls. Genomic DNA was obtained from peripheral blood and polymorphisms were investigated using restriction fragment length polymorphism analysis (RFLP). There were no significant differences in genotype frequencies of FSH receptor gene between endometriosis patients and controls. For the heterozygous type of the FSH receptor polymorphism (c.-29G>A) we did not find a significant difference in its frequency between patients with minimal/mild and moderate/severe endometriosis (p = 0.136). Also, the FSH beta chain (c.-280G> T) polymorphism frequency was not significantly associated with the severity of endometriosis (p = 0.966). FSH receptor and FSH beta chain polymorphisms do not seem to influence the severity of endometriosis, but they could be correlated with female infertility (primary or secondary), therefore further studies are required to debate this topic.

The regulatory effect of genistein on P450 aromatase and follicle-stimulating hormone receptor in mouse experimental model of menopausal metabolic syndrome.

In order to investigate the mechanism of genistein (Gen) in the treatment of climacteric syndrome, an in vivo study was performed to investigate the beneficial effects of genistein on the expression of P450 aromatase (P450 arom) and follicle-stimulating hormone receptor (FSHR) in the mouse ovary and uterus. Fifty female ICR mice (45±5g, n=50), aged 12months, were divided into the following five groups with 10 animals in each: blank control group (CG), low-dose genistein group (L-Gen), middle-dose genistein group (M-Gen) and high-dose genistein group (H-Gen) (received 15, 30 and 60mg/kg of genistein, respectively), and oestrogen group (EG; received 0.5mg/kg diethylstilbestrol). The expression levels of the FSHR protein were determined by an immunohistochemical staining method. The expression of P450 arom, Cytochrome P450 19 (CYP19) and FSHR was quantified by real-time PCR. Immunohistochemical results showed that the expression levels of the FSHR protein in the M-Gen (average stained area: 20.79) and the H-Gen (average stained area: 21.21) groups were significantly stronger than in the CG (average area was 17.24) group (p<.05). The expression levels of CYP19 mRNA and P450 arom were positively correlated with the dose of genistein. Specifically, the relative expression levels in the H-Gen and EG groups were more than 1.5 times higher than in the CG group (p<.05). Genistein played a significant role in regulating aromatase and FSHR gene expression to improve perimenopausal ovarian and uterine function.

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.

Relationship of follicle stimulating hormone receptor Gene Thr307Ala polymorphism with polycystic ovary syndrome in Iraqi women

Polycystic ovary syndrome (PCOS) is a polygenic multifactorial status affecting millions of females worldwide. It is a common cause of anovulatory subfertility. Because of follicle stimulating hormone (FSH), is an important agent in human reproduction. Therefore, the correlation between follicle stimulating hormone receptor (FSHR) gene polymorphisms and polycystic ovary syndrome attracts broad attention. The objective of this study is to investigate the potential association between the follicle stimulating hormone receptor gene Thr307Ala polymorphism with polycystic ovary syndrome in the Iraqi women. A case-control study including 135 Iraqi women of Arab ethnicity (75 PCOS patients and 60 age-matched control women). The age of subjects ranged from 18 to 38 years. PCOS diagnosis was established by Rotterdam consensus criteria. The FSHR (Thr307Ala) variant was tested by conventional polymerase chain reaction (PCR)–restriction fragment length polymorphism (RFLP) followed by deoxyribonucleic acid (DNA) sequencing. The heterozygote Thr/Ala (AG) genotype of Thr307Ala (rs6165) polymorphism of follicle stimulating hormone receptor gene was giving a significant risk (odds ratio=19.4, 95%CI=1.14-30.40, P value=0.002) of developing PCOS in Iraqi women compared with control group. Sequencing analysis of DNA confirms RFLP analysis. In conclusion; the variant Thr307Ala (rs6165) of the follicle stimulating hormone receptor gene is associated with polycystic ovary syndrome and may consider as the causal factor of polycystic ovary syndrome in Iraqi women.

Pilot study: Alternative spliced variants of the Follicle-Stimulating Hormone Receptor gene in Greek women undergoing In Vitro Fertilisation (IVF)

Assisted Reproduction Technology (ART) has proven to be a valuable tool for infertile couples. Unfortunately, no perfect ovarian stimulation protocol has been designed and ovarian response shows great variability among women. Pharmacogenomics aims at detecting genetic markers so as to individualize protocols in order to maximize ovarian response to treatment. Follicle Stimulation hormone receptor (FSHR) mutations and single nucleotide polymorphisms have been extensively studied. Splice variants of the FSHR have been detected in women undergoing in vitro fertilisation (IVF). This study aims to determine the presence FSH splice variants in Greek women undergoing IVF. RNA was extracted from cumulus cells from 35 women and analysed by real time PCR. Splice variants were detected by gel electrophoresis. Three cases of deletion of exon 9 and 2 cases of insertion of intron 8 were detected in our study group. No association between the presence of splice variants and response to ovarian stimulation was detected. Two live births were detected one in each variant group. Even though these two types of splice variant detected do not show any clinical correlation it is believed that variants of the FSHR may be associated with poor or high response to exogenous gonadotrophin so further research is necessary.

Cloning and expression analysis of the follicle-stimulating hormone receptor (FSHR) gene in the reproductive axis of female yaks (Bos grunniens)

Follicle-stimulating hormone receptor (FSHR) plays a central role in promoting follicle maturation through the follicle-stimulating hormone (FSH)-mediated cAMP pathway in animals. The objectives of the present study were to clone the FSHR gene of yaks (Bos grunniens) and compare differences in FSHR mRNA expression in the reproductive axis between yaks and cattle. Hypothalamus, anterior pituitary, oviduct, ovary, and uterus tissue samples were collected from adult female yaks (n = 5) and cattle (n = 5) during the follicular phase. Using reverse transcriptase-polymerase chain reaction (RT-PCR), we found that the FSHR coding region of the yak is 2088 bp and encodes 695 amino acids. Its amino acid sequence showed 99.38%–72.22% similarity to the homologous genes of cattle, goats, sheep, cats, donkeys, horses, humans, chickens, monkeys, mice, rats, and wild boar. Real-time PCR analysis revealed that the FSHR gene was expressed in all tissues examined. Expression of the FSHR gene in the yak was higher in the uterus than other tissues (P < 0.05) but, in cattle, was higher in the ovary than other tissues (P < 0.05). The FSHR gene expression level in the cattle ovary was significantly higher than that in the yak ovary (P < 0.01). These results indicate that the FSHR gene is relatively conserved in the course of animal evolution. The variation in sequence and expression level of FSHR between the two species might be associated with the difference in their reproduction.

Follicle stimulating hormone receptor gene polymorphism and its association with fecundity in goats

Pluriparous goats (73) in between second to fifth parity with history of breeding 30 days before were selected for present study. A real time B-mode ultrasonography machine equipped with a linear array 7.5 MHz transrectal scanner was used for diagnosis of early pregnancy and twinning in goats. Out of 73 goats, 40 were detected pregnant with 22 goats with single embryo and 18 goats with twin embryos. Genomic DNA was extracted from the blood of all pregnant goats and polymerase chain reaction (PCR) with FSHR gene specific primers was carried out. The PCR product for FSHR gene was digested by restriction enzyme MSCI. Two genotypes AA and AB having amplification size (214 bp, 90 bp) and (214 bp, 90 bp and 304 bp) were detected, respectively. In AA genotype, 21 single and 9 twins were observed while 1 single and 9 twin embryos were observed in AB genotype. The percentage of single and twins was 70% and 30% in AA genotype, and 10% and 90% in AB genotype. In AA genotype, the average litter size was 1.3 while in AB genotype it was 1.9 and the difference was statistially significant. The AB genotype for the FSHR gene is responsible for more twining percentage.