Luteinizing Hormone/choriogonadotropin Receptor Gene Research Articles

Gene expression by equine oocytes and cumulus cells, a window into the unknown

In vitro maturation (IVM) of oocytes (OC) is a critical step for the in vitro production of equine embryos. However, the regulatory mechanisms, biological processes, cellular components, and molecular functions involved in IVM are not entirely understood. We previously analyzed the gene expression dynamics of gonadotropin receptors in cumulus cells (CC) during IVM, demonstrating an upregulation of the follicle-stimulating hormone receptor gene (FSHR) and downregulation of the luteinizing hormone choriogonadotropin receptor gene (LHCGR) in mature compared to immature CC (de la Fuente.et.al. Journal of Equine Veterinary Science 2022; 113). Moreover, we found that CC from in vivo maturing cumulus-oocyte complexes (COC) obtained from dominant gonadotropin-stimulated follicles (DF-CC) had a lower expression of FSHR and a higher expression of LHCGR compared to their in vitro counterparts. However, the effect of IVM medium composition on the expression of gonadotropin receptors has not been investigated. Thus, the aim of this study was to determine differences in the gene expression of OC and CC following IVM of three reported IVM media: A) EQ-IVM (IVF-Bioscience) a commercially available medium, B) DMEM/F12-based medium (Meyers.et.al. Reproduction, Fertility and Development. 2019; 31:1874-1884.) and C) M199-based medium (Choi.et.al. Theriogenology. 2002; 58:771-774) and their difference with DF-CC samples. Extracted total RNA from OC and CC was sequenced and raw reads were trimmed (trimmgalore) and mapped (STAR) to the current equine referencegenome. Mapped reads were quantified (featurecount) and compared between groups, using the DESeq2 package (FDR cutoff 0.1; Min (log2) fold change 1.5). Overall, there was a mapping rate of >85%, and an average of 14,000 expressed genes in OC and CC samples with ∼68% categorized as protein-coding genes. Based on gene expression dis-similarities, OC and CC samples were partially clustered according to their respective maturation media, but all samples showed a different gene expression profile compared to DF-CC samples. Comparing the gene expression of CC between the A and B media, we found 607 differentially expressed genes (DEGs), 1905 DEGs between A and C, and 957 DEGs between B and C. All three media showed a similar DEGs number when compared to DF-CC samples (∼10K DEGs). FSHR showed a higher expression in all IVM samples compared to DF-CC. However, LHCGR presented a lower expression only in two of the IVM media used (B & C) compared to DF-CC, showing no significant expression difference in the commercial IVM media tested. These data highlight the dynamics of gonadotropin receptors during IVM, which may contribute to the understanding of the physiology of oocyte maturation and allow optimization of IVM conditions in horses. Funding: The Theriogenology Foundation, the Population Health and Reproduction Department Seed Grant, and the Center for Equine Health.

A rare cause of primary amenorrhea: LHCGR gene mutations

IntroductionThe luteinizing hormone/choriogonadotropin receptor (LHCGR) plays a critical role in sexual differentiation and reproductive functions in men and women. Inactivating mutations in this gene lead to Leydig cell hypoplasia (LCH), and cause disorders of sex development (DSD) in patients with 46,XY. In this study, it was aimed to discuss the clinical, laboratory and molecular genetic analysis results of nine patients with 46,XY karyotype who had mutations in the LHCGR gene. Materials and methodsThe ages, complaints, anthropometric measurements and hormonal results (follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone) of the patients at the time of admission were recorded retrospectively from their medical records. The mutations in the LHCGR gene were investigated using the Sanger sequencing method. FindingsIn this study, LHCGR gene mutations were detected in a total of nine patients as a result of the analysis of the index patients presenting with primary amenorrhea from four different families and the examination of the families. In the first three families with no consanguinity between, the same mutation was detected in seven patients in total (Homozygous c.161 + 4A > G). A different mutation was detected in the fourth family (Homozygous p.A483D c.1448C > A). ConclusionIn this study, nine patients with karyotype 46,XY, most of whom presented with the complaint of delayed puberty/primary amenorrhea, were diagnosed with LCH. Especially in patients, in whom the elevation of LH is pronounced and there is no testosterone synthesis, LCH should be considered.

Effect of LHCGR and FSHR gene polymorphisms on fertility traits and milk yield of cross-bred dairy cows in Sri Lanka

Single nucleotide polymorphism (SNP) analysis of fertility genes will be useful in improving the genetic potential of dairy cows. A population of 142 cross-bred dairy cows was screened for SNPs in bovine luteinizing hormone choriogonadotropin receptor (LHCGR) and bovine follicle stimulating hormone receptor (FSHR) genes. The effect of reported SNPs on selected fertility traits (average calving interval, average number of services per conception and age at first calving) and milk yield was determined. Altogether six SNPs were detected in animals screened under the present study. Among them, four SNPs (rs41256848, rs41256850, rs465790244, and rs45463781) were located in the exon 11 region of the LHCGR gene and two SNPs (rs43676359 and G-278-A (GU253337) were located in the five upstream region of the FSHR gene. Minor alleles identified for SNPs in the FSHR gene in the studied small population of cows differed from those reported in other research. In this study, only rs45463781 SNP at exon 11 of the LHCGR gene significantly affected the average milk yield in cross-bred cows. The nucleotide change from “G” to “A” negatively affected the average milk yield. Further investigations are needed to confirm the reported association with a larger sample size.

P–534 A novel heterozygous mutation in the luteinizing hormone/choriogonadotropin receptor (LHCGR) gene in a patient with ‘genuine’ empty follicle syndrome

Abstract Study question Whether empty follicle syndrome (EFS) in a patient has a genetic basis. Summary answer Our findings would expand the mutational spectrum of LHCGR, in patients with GEFS What is known already The LHCGR gene (OMIM #52790) is located on chromosome 2p21 has 11 exons The LHCGRs present in gonadal cells; granulosa, theca and luteal cells in women and Leydig cells in menandplays a critical role in male sexual differentiation, female ovarian development and fertility (folliculogenesis, ovulation, corpus luteum formation and progesterone secretion) Inactivating mutations in males can lead to Leydig cell hypoplasia, which causes disorders in sexual development (MIM #238 320). Phenotype of women is less severe and variable and has no effect on the secondary sex characteristics, but it could cause amenorrhoea and infertility Study design, size, duration: In the context of clinical genetics, Next Generation Sequencing libraries were prepared in line with the manufacturer’s orders using QIASeq™ Targeted DNA Custom Panel (Qiagen) targeting exons and 20 bp exon-intron boundaries of selected genes (AR, BMP15, CATSPER1, CFTR, CYP21A2, FSHB, FSHR, HESX1, LHB, LHCGR, NR5A1, POU1F1, SRY, ZP1). The variant detected in NGS analysis was further confirmed using Sanger sequencing (MiSeq, Illumina, San Diego, CA). Participants/materials, setting, methods A 27 years old Turkish women with 6 year history of primary unexplained infertility underwent controlled ovarian hyperstimulation and IVF with an antagonist protocole in Ankara City Hospital. Although normal follicular development, E2 levels, and bioavailable β-hCG plasma levels, no oocytes or cumulus-corona complexes were retrieved by follicular aspiration. Main results and the role of chance A novel heterozygous mutation on Exon 5 of LHCGR (NM_000233.4):c.453C>G (p.Phe151Leu). This variant has not been reported in GnomAD database and is a novel variant as per controlled from ClinVar and HGMD mutation databases Also coagulation tests were done Patient was heterozygote for the prothrombin mutation (FXIII) and homozygote for MTHFR C677T mutation. The patient has normal pubertal development and female karyotype (46,XX). Gonadotropin and E2 levels were normal, nor any history of anosmia, primary amenorrhoea, polycystic ovaries, hyperandrogenism, systemic disorder, or neurologic defect. According to ACMG 2015 and HGMD detected variant was classified as unknown clinical significance (VUS) variant In 22nd World Congress COGI in 2015 we have presented another recurrent GEFS case with compound heterozygous frameshift mutations in exon 5 and 11 of LHCGRgene(c.1764_1765insT) and (c.430G>T, p.V144F) who developed premature ovarian failure at the end The different types of LHCGR mutations would lead to the different functional effects and clinical phenotypes in different ages; primary amenorrhea with high levels of LH, poor ovarian response (Poesidon group1), GEFS and secondary amenorrhea (premature ovarian failure) GEFS may be a gradual biological occurrence related to ovarian ageing. Long term follow up is important, since some clinical manifestations appear later in life. Limitations, reasons for caution LH resistance in females were always found due to their effected brothers and only 10 GEFS cases of 46, XX females with LHCGR gene defect reported in literature. Patient was heterozygous for indicated mutation; therefore segregation study should be done in family members and in vitro studies should be performed. Wider implications of the findings: Screening for mutations in the LHCGR gene in Poseidon group 1 and in patients GEFS especially the recurrent ones will provide valuable information and time for clinical management, corresponding treatment and give the opportunity of wise and cost effective counseling of patients about their future reproductive choices. Trial registration number it is a case report

Association between Luteinizing Hormone/Choriogonadotropin Receptor Ins18LQ Gene Polymorphism and Polycystic Ovary Syndrome

BACKGROUND: Polycystic ovary syndrome (PCOS) is one of the most common endocrine disorders of women in the childbearing period. However, its pathophysiology is still unclear. Certain polymorphisms of the luteinizing hormone/choriogonadotropin receptor (LHCGR) genes may lead to changes in the bioactivity of this hormone. The important functional role of LHCGR in the metabolism of androgen and ovulation, the LHCGR gene variant, may be related to the risk of PCOS.
 AIM: The aim of this study was to evaluate the association between LHCGR Ins18LQ gene polymorphism and PCOS.
 METHODS: A case–control study was performed in women with PCOS and non-PCOS from May 2019 to October 2019 in HFC IVF Center. We included 50 women with PCOS and 50 healthy controls. Polymorphism of the LHCGR (ins18LQ) gene was genotyped using polymerase chain reaction-restriction fragment length polymorphism.
 RESULTS: From this study, we found that there was no significant difference in the proportion of ages between the groups (p > 0.05). There were significant differences in the characteristics of body mass index, FSH level, LH level, and LH/FSH ratio between the PCOS and control groups (p < 0.05). We also found that the proportion of heterozygote variant non-ins/ins was higher in the PCOS group compared to the control group, but there was no significant difference between the polymorphisms of the non-ins and non-nonins variants between the PCOS and control groups (p = 0.269). The frequency of ins alleles was higher in the PCOS group compared to the control group.
 CONCLUSION: There was no significant association between LHCGR ins18LQ gene polymorphism and PCOS.

Frequencies of genotypes and alleles of rs41256848 in the LHCGR gene in the population of black-and-white holsteinized cattle

Embryo transfer technique is one of the key in accelerated reproduction of cattle. One of the most important stages is the selection of donor cows that are most sensitive to the procedure of hormonal stimulation of ovulation. One of the promising genetic markers of the reproductive status of cattle is currently the gene encoding the luteinizing hormone / choriogonadotropin receptor (LHCGR). One of the SNP in the LHCGR gene of cattle has already been described in the literature as associated with the number of oocytes and the quality of embryos - rs41256848 (c.1401G> T, p.Trp467Cys). The purpose of this work was to estimate the frequencies of genotypes and alleles of this substitution in the population of Black-and-White holsteinized cattle (190 cows). Genotyping was performed by PCR-RFLP method. In the studied population of cattle, the frequency of the G allele associated with higher rates in the total number of oocytes and the number of embryos survived after transplantation, as well as with the least number of unfertilized oocytes, is 63.2%.

SUN-LB052 Sexual Precocity in a Male Toddler Caused by Inadvertent Prolonged Exposure to Exogenous Testosterone

Background: Unintentional exposure to cutaneous androgen products leading to virilization has previously been described in children, however, continues to be an overlooked cause of precocious sexual development. Clinical Case: A 16-month-old boy presented for a second opinion consultation regarding sexual precocity. Parents noticed enlargement of his penis and appearance of sparse pubic hair at about 4 months of age, prompting referral to an endocrinologist by 10 months. A comprehensive evaluation was performed. The bone age was advanced at 13-15 months and his total testosterone was 243 ng/dL (normal < 20 ng/dL). An ACTH stimulation test was negative for disorders of steroid biosynthesis. Ultrasound examination showed normal testes and no evidence of adrenal or hepatic masses. A Lupron stimulation test, performed at 14 months of age, showed a peak LH of 1.41 mIU/mL (normal < 5 mIU/mL) and peak total testosterone of 21 ng/dL (reference range ≤ 5 ng/dL). The patient was diagnosed with familial gonadotropin-independent male-limited sexual precocity, a rare condition caused by activating mutations of the LHCGR (luteinizing hormone/choriogonadotropin receptor) gene. Treatment with bicalutamide and anastrozole was prescribed. Owing to parental concern about side effects of the prescribed medications, they did not start the recommended treatment and sought a second opinion. On exam, his length was > 99th percentile for age, stretched penile length 7.5 cm (mean ± standard deviation for this age: 4.82 ± 0.44 cm), mid-shaft diameter 2 cm, and testes 2 mL bilaterally. The scrotum was thin and pendulous and there were a few pubic hairs at the base of the penis. His total testosterone was < 10 ng/dL, LH < 0.1 mIU/mL, and hCG < 0.1 mIU/mL (all normal). Ultrasound of the testes was normal. After the visit, a more thorough history was obtained from the mother. She remembered that the child’s maternal grandfather, who provided direct care to the patient in the first year of life while she attended school, used transdermal testosterone gel daily for treatment of hypogonadism. Since the grandfather’s absence after the patient’s first birthday, the child has not had any further progression of his secondary sex characteristics. On follow-up visit at 17 months of age, patient’s exam was stable. Genetic testing for mutations of the LHCGR gene was negative. Conclusion: Exogenous exposure to transdermal androgens should be kept high on the differential diagnosis for children presenting with evidence of peripheral sexual precocity as it can save patients unnecessary and costly work-up and intervention. Physicians should also be aware of their own anchoring biases, especially when evaluating patients for a second opinion of rare diagnoses. Lastly, adult endocrinologists should be mindful in counseling patients prescribed topical testosterone agents about the risk of cutaneous exposure to female partners and children. Unless otherwise noted, all abstracts presented at ENDO are embargoed until the date and time of presentation. For oral presentations, the abstracts are embargoed until the session begins. s presented at a news conference are embargoed until the date and time of the news conference. The Endocrine Society reserves the right to lift the embargo on specific abstracts that are selected for promotion prior to or during ENDO.

Association of luteinizing hormone/choriogonadotropin receptor gene polymorphisms with polycystic ovary syndrome risk: a meta-analysis

To investigate the association between Luteinizing hormone/choriogonadotropin receptor (LHCGR) gene polymorphisms and polycystic ovary syndrome (PCOS). A systematic literature search and meta-analysis using STATA software for included studies. Fourteen case-control studies containing rs13405728, rs4539842, and rs2293275 of LHCGR gene were included, which was comprised of 11,738 PCOS cases and 35,329 controls. Results of the meta-analysis showed a significant association between PCOS and rs13405728 (for G vs. A: OR = 0.735, 95% CI = 0.699–0.773, p<.001; For GG vs. AG + AA: OR = 0.578, 95% CI = 0.436–0.767, p<.001; For GG + AG vs. AA: OR = 0.817, 95% CI = 0.741–0.901, p<.001) in Asian populations, and rs4539842 (for ins/ins vs. ins/non + non/non: OR = 0.686, 95% CI = 0.483–0.974, p=.035) and rs2293275 (for AA vs. AG + GG: OR = 4.115, 95% CI = 1.033–16.38, p=.045) in Caucasian populations, respectively. LHCGR gene variations are population specifically associated with PCOS, which indicated these SNPs in LHCGR may contribute to the pathogenesis of PCOS and could be used as potential biomarkers to predict the risk of PCOS.

Cosegregation of a novel mutation in the sixth transmembrane segment of the luteinizing/choriogonadotropin hormone receptor with two Brazilian siblings with severe testotoxicosis

ABSTRACTPurpose: Testotoxicosis is an autosomal dominant form of gonadotropin-independent precocious puberty caused by heterozygous constitutively activating mutations of the luteinizing hormone/choriogonadotropin receptor (LHCGR) gene. The aim of this study was to describe two Brazilian siblings with testotoxicosis, to confirm the molecular diagnosis, and to perform an in silico analysis of a novel mutation in the hot spot of the LHCGR gene. Materials and methods: Molecular analysis of the mutation on the LHCGR gene was performed by direct Sanger sequencing, followed by an in silico analysis using HOPE bioinformatics tool to predict a functional defect of the mutant. Results: Both patients presented with gonadotropin-independent precocious puberty before the age of four years. Genetic analysis revealed a novel non-maternally inherited p.Asp578Val mutation of the LHCGR gene. An in silico analysis showed that the p.Asp578Val mutation disturbed amino acid physicochemical features regarding its size, charge, and hydrophobicity value. Conclusions: Clinical and hormonal profile of the siblings here evaluated was not different while compared to those patients previously described. An in silico mutation analysis reinforced the causative role of recurrent activating mutations in the intracellular loop and transmembrane helices of the LHCGR. The segregation of this mutation with the offsprings’ phenotype indicated that it is causative.

Association of the luteinizing hormone/choriogonadotropin receptor gene polymorphism with polycystic ovary syndrome

This study aimed at evaluating possible associations of the single nucleotide polymorphism (SNP) in luteinizing hormone/choriogonadotropin receptor (LHCGR) gene G935A and polycystic ovary syndrome (PCOS) phenotype. The study included 100 PCOS female patients and 60 healthy female control subjects. The patients were recruited from the Gynecology out-patient clinic, Kasr Al-Aini Hospital, Cairo University. All candidates underwent full history taking and clinical examination with calculation of body mass index. Serum and EDTA samples were collected from each patient after a written consent. A hormonal profile was done for each patient as well as DNA analysis of the G935A polymorphism of LHCGR gene. In PCOS group, 26% were homozygous (AA), 27% were heterozygous (GA) and 47% were wild genotype (GG), while in controls 30% were heterozygous and 70% were wild genotype (OR: 2.25; CI: 1.16–4.386; p value: 0.012). The homozygous 935A individuals were at higher risk to develop PCOS than controls (OR: 1.80; CI: 1.54–2.09; p value < 0.001).We found a genetic variant, which is associated with PCOS in a sample of the Egyptian population. These results may provide an opportunity to test this SNP at the LHCGR gene in fertile or infertile women with family history to assess their risk of PCOS.

Effect of luteinizing hormone/choriogonadotropin receptor (LHCGR) gene on chicken reproductive traits

Luteinizing hormone/choriogonadotropin receptor (LHCGR) gene, potentially related to reproductive traits in chickens, was genotyped by using the Pooled DNA Sequencing, PCR-SSCP and Directing Sequencing techniques. 306 Erlang Mountain chickens form one line (SD03, a line that has been selected for egg quality from a local chicken breed in Sichuan province, China) were genotyped in this study. The associations between LHCGR polymorphisms and six reproductive traits [body weight at first egg (BWAFE), weight of first egg, age at first egg (AFE), number of eggs at 300days of age (EN), body weight at 300days of age and egg weight at 300days of age (EWTA)] were estimated using the one-way analysis of variance method. Results showed that SNP +G4058A and SNP +T4099G of the LHCGR gene were significantly associated with BWFE and AFE. Birds with the AG genotype for the +G4058A SNP exhibited shorter AFE (P<0.05) and greater EN than those of the GG and AA genotypes, suggesting a balancing selection (overdominance); the effect of allele C in SNP +C3021T and allele C in SNP +T4490C on EN and AFE is additive and may reflect the influence of positive selection. These alleles have promise as genetic markers for future marker-assisted selection.

Association of a missense mutation in the luteinizing hormone/choriogonadotropin receptor gene (LHCGR) with superovulation traits in Chinese Holstein heifers

BackgroundUpon binding luteinizing hormone in the ovary, the luteinizing hormone/choriogonadotropin receptor (LHCGR) is necessary for follicular maturation and ovulation, as well as luteal function. We detected mutations in the LHCGR gene and evaluated their association with superovulation.MethodsUsing polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and DNA sequencing, we examined polymorphisms in LHCGR and the genotypes associated with superovulation traits in 127 Chinese Holstein heifers.ResultsA G/T polymorphism (ss52050737) in exon 11 was significantly associated with the total number of ova and the number of transferable embryos.ConclusionsLHCGR may be a new predictor for superovulation in Chinese Holstein heifers.

Polymorphisms of the bovine luteinizing hormone/choriogonadotropin receptor (LHCGR) gene and its association with superovulation traits

The major limitation to the development of embryo transfer technique in cattle is the highly variable between individuals in ovulatory response to FSH-induced superovulation. The objective of this study was to identify a predictor to forecast superovulation response on the basis of associations between superovulation performance and gene polymorphism, variation in the bovine luteinizing hormone/choriogonadotropin receptor (LHCGR) gene was investigated using PCR-single-strand conformational (PCR-SSCP) and DNA sequencing. Four single nucleotide polymorphisms (SNPs) of G51656T, A51703G, A51726G and G51737A were identified at the intron 9 of the LHCGR gene in 171 Chinese Holstein cows treated for superovulation, and evaluated its associations with superovulatory response. Association analysis showed that these four SNPs had significant effects on the total number of ova (TNO) (P < 0.05). Moreover, the A51703G and A51726G polymorphisms significantly associated with the number of transferable embryos (NTE) (P < 0.05). In addition, significant additive effect on TNO was detected in polymorphisms of G51656T (P < 0.05) and A51703G (P < 0.01), and the A51703G polymorphism also had significant additive effects on NTE (P < 0.01). These results indicate that LHCGR gene is a potential marker for superovulation response and can be used to predict the most appropriate dose of FSH for superovulation in Chinese Holstein cows.

A Case of Male-Limited Precocious Puberty Caused by a Point Mutation in the Second Transmembrane Domain of the Luteinizing Hormone Choriogonadotropin Receptor Gene

We describe a Japanese patient with male-limited precocious puberty who has a heterozygous thymine to cytosine (T to C) transition at nucleotide 1193; the mutation encodes a methionine to threonine substitution in residue 398 (M398T) of transmembrane helix 2 of the luteinizing hormone/choriogonadotropin receptor. Transfected into COS-7 cells, M398T exhibited constitutively high basal cAMP levels but retained an agonist-induced cAMP response. The constitutively higher cAMP levels caused by M398T are consistent with Leydig cell activation and precocious puberty in the patient. By comparison to wild type receptor, M398T transfectants have significantly lower agonist-induced inositol phosphate (IP) levels at >10 −10M hCG concentrations and a higher apparent affinity for binding hCG. These data suggest that the M398T substitution alters Gq coupling and phospholipase-C activation, as well as G scoupling and adenylyl cyclase activity, and changes the conformation of the extracellular domain of the receptor.