Eur J Endocrinol
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

DOI: 10.1530/eje.1.02031
European Journal of Endocrinology, Vol 153, Issue 6, 845-852
Copyright © 2005 by European Society of Endocrinology
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via ISI Web of Science (20)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Lanfranco, F.
Right arrow Articles by Simoni, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Lanfranco, F.
Right arrow Articles by Simoni, M.
CLINICAL STUDY

Role of sequence variations of the GnRH receptor and G protein-coupled receptor 54 gene in male idiopathic hypogonadotropic hypogonadism

Fabio Lanfranco1,2, Jörg Gromoll1, Sigrid von Eckardstein1, Eva M Herding1, Eberhard Nieschlag1 and Manuela Simoni1

1 Institute of Reproductive Medicine of the University, D-48129 Münster, Germany and 2 Division of Endocrinology and Metabolism, Department of Internal Medicine, University of Torino, Italy

(Correspondence should be addressed to E Nieschlag; Email: Eberhard.Nieschlag{at}ukmuenster.de)


   Abstract
Top
 Abstract
Introduction
 Subjects and methods
 Results
 GnRHR gene
 GPR54 gene
 Discussion
 References
 
Objective: To determine the frequency of mutations of the gonadotropin-releasing hormone receptor (GnRHR) and of the G protein-coupled receptor 54 (GPR54) genes in normosmic idiopathic hypogonadotropic hypogonadism (IHH).

Methods: In a retrospective study we analyzed the GnRHR and the GPR54 genes of 45 IHH patients and 50 controls. Genomic DNA was amplified by PCR to obtain partially overlapping amplicons encompassing the exon–intron boundaries of the GnRHR and GPR54 genes and analyzed by single-stranded conformation polymorphism gel electrophoresis and/or DNA sequencing.

Results: One heterozygous R262Q mutation of the GnRHR gene was identified in one patient with familial IHH. The silent single-nucleotide polymorphism (SNP) 453C > T occurred at the same frequency in patients and controls. One patient with sporadic IHH and consanguineous parents showed a novel homozygous sequence variation of the GPR54 gene (1001_1002insC) resulting in an open reading frame shift and elongation of 43 amino acids with an increased number of proline residues in the intracellular receptor domain. This patient had delayed puberty, low testosterone (3.4 nmol/l), and low-normal LH and FSH levels responsive to GnRH. Pulsatile GnRH administration normalized testosterone levels and induced spermatogenesis sufficiently to induce a pregnancy with assisted reproduction. Two common SNPs in exon 1 and exon 5 of the GPR54 gene showed similar frequency distribution and hormonal profiles in IHH and controls.

Conclusions: Mutations of the GnRHR and of the GPR54 gene are rare in IHH and should be investigated especially in cases with autosomal recessive transmission. Common SNPs of the GnRHR and GPR54 genes do not play any role in IHH.


   Introduction
Top
 Abstract
 Introduction
Subjects and methods
 Results
 GnRHR gene
 GPR54 gene
 Discussion
 References
 
Idiopathic hypogonadotropic hypogonadism (IHH) is a genetically heterogeneous condition characterized by a functional deficit in hypothalamic gonadotropin-releasing hormone (GnRH) production or action (1–3). Patients with IHH have been classically divided into those with anosmia or hyposmia (Kallmann syndrome) and those with normal olfaction (4, 5). Sporadic and familial cases with different modes of inheritance have been described. Mutations in two distinctive genes located at the short arm of the X-chromosome, KAL1 and DAX1, are responsible for the X-linked forms of IHH (2, 4–8). Mutations of the fibroblast growth factor receptor 1 gene were recently reported in patients with Kallmann syndrome (9, 10). These were the only identified causative genes involved in inherited human IHH, and they were dominant. The molecular basis of the first autosomal recessive form of IHH, affecting both women and men, was determined to be mutations of the GnRH receptor (GnRHR) gene (11, 12).

GnRHR is a G protein-coupled receptor with seven transmembrane segments and an extracellular amino terminus but no intracellular carboxyl terminus; it activates phospholipase C and mobilizes intracellular calcium via G proteins (13, 14). The GnRHR gene (NM_000406 [GenBank] ) is localized on chromosome 4 at band q13.1 (15). The coding sequence comprises three exons and spans over 20 kb. Numerous compound heterozygous (11, 12, 16–18) and four homozygous (19–22) GnRHR mutations have been described, causing subtypes of IHH varying from complete to partial resistance to GnRH.

GnRHR mutations account for less than 50% of familial IHH cases and a small fraction of sporadic cases, indicating that IHH is a heterogeneous condition caused by various genetic defects. A new cause for IHH has been recently described: it consists of mutations of the G protein-coupled receptor 54 (GPR54) gene, which is localized on the short arm of chromosome 19 (19p13). GPR54 (NM_032551 [GenBank] ) is a G protein-coupled receptor expressed mainly in the brain, pituitary and placenta. GPR54 is formed of five exons and encodes a 398 amino acid protein. In the past 2 years homozygozity mapping and candidate gene analysis of large consanguineous pedigrees led two groups to describe loss-of-function mutations in GPR54 in association with autosomal recessive IHH (23, 24). In parallel, the independent generation and characterization of a Gpr54 knockout mouse model, which shows a phenocopy of human isolated IHH, demonstrated that the function of GPR54 is conserved in mammals and that GPR54 is necessary for the normal function of this axis (24, 25). A further case of GPR54 compound heterozygous mutation in a patient with IHH was described recently (26).

The aim of this study was to analyze the occurrence of sequence variants, mutations and single-nucleotide polymorphisms (SNPs) of the GnRHR and GPR54 gene in a cohort of 45 male patients with normosmic IHH. We report the clinical phenotype of a subject with consanguineous parents who showed a novel homozygous mutation in the GPR54 gene and the SNP analysis of the GnRH and GPR54 genes in hypogonadal patients and controls.


   Subjects and methods
Top
 Abstract
 Introduction
 Subjects and methods
Results
 GnRHR gene
 GPR54 gene
 Discussion
 References
 
Subjects

In a retrospective study design, 45 hypogonadal male patients were selected from the database of the outpatient clinic of the Institute of Reproductive Medicine of the University in Münster, Germany. Six of the 45 patients reported at least another case of IHH, defined as lack of spontaneous pubertal maturation requiring medical intervention, in their family (familial cases). However, since detailed pedigrees were not available, the mode of inheritance could not be identified. The other 39 patients were sporadic cases. Hypogonadotropic hypogonadism was defined as testosterone concentrations below 12 nmol/l in the setting of inappropriately low gonadotropin levels (below or around the lowest limit of the normal range: 2 IU/l for luteinizing hormone (LH) and 1 IU/l for follicle-stimulating hormone (FSH) upon repeated determinations. Additional elements for the diagnosis of IHH were: absence of spontaneous pubertal maturation, no evidence of a hypothalamic–pituitary anatomical lesion on imaging, and otherwise normal pituitary function as documented by normal basal and stimulated levels of prolactin, thyroid-stimulating hormone, growth hormone and cortisol. All patients were normosmic, as assessed by a smell test in which the subject was asked to identify odors produced by aromatic essences. Eight patients had bilateral and five patients monolateral undescended testes. Patients provided a blood sample for DNA analysis and clinical data were retrieved from the database and patient charts. The control group consisted of 50 eugonadal men with normal hormone levels and normal semen analysis recruited as volunteers to participate in a male contraception trial. All subjects were of Caucasian origin. Patients and controls gave written informed consent according to the protocol approved by the Ethics Committee of the Medical Faculty and State Medical Board.

Genetic screening

Genomic DNAwas extracted from peripheral blood using the FlexiGene DNA extraction kit (Qiagen, Helden, Germany) according to the manufacturer’s instructions.

Mutation screening of the GnRHR and of the GPR54 gene were performed by single-stranded conformation polymorphism (SSCP) gel electrophoresis as previously described (27, 28). Genomic DNA was amplified by PCR to obtain partially overlapping amplicons encompassing the exon–intron boundaries. The primers used are shown in Tables 1Go and 2Go. The amplicons were further analyzed by SSCP. Fragments showing an abnormal migration pattern were sequenced. In addition, about one-third of the amplicons randomly chosen among those with a normal migration pattern were directly sequenced in order to check the accuracy of the SSCP results. SSCP analysis and subsequent sequencing revealed common SNPs both in the GnRHR gene (453C > T, refSNP ID: rs4986942) and in the GPR54 gene (24A > G, refSNP ID: rs10407968 and 1091A > T, refSNP ID: rs350132). The GnRHR 453C > T SNP was further screened by MaeIII restriction fragment length analysis. The SNPs of the GPR54 gene were analyzed by SSCP and sequencing.


View this table:
[in this window]
[in a new window]
  		Table 1 Primers used in the anlaysis of the GnRHR gene.
 

View this table:
[in this window]
[in a new window]
  		Table 2 Primers used in the analysis of the GPR54 gene.
 
Hormone measurements

Serum hormones were analyzed as described previously (27, 29). Serum LH, FSH, estradiol and sex hormone binding globulin (SHBG) were assayed by fluoroimmunoassay using Autodelfia equipment (Perkin Elmer, Freiburg, Germany). Testosterone levels were determined using a commercial ELISA (DRG, Marburg, Germany). Intra- and inter-assay coefficients of variation were below 10%. The normal range for unextracted serum testosterone is 12–35 nmol/l. All analytical methods were performed and documented in accordance with the ISO9001 certified quality management system applied in our laboratory.

Statistics

After exclusion of the two patients with mutations of the GnRHR and of the GPR54 gene, SNP data of 43 patients were considered. In addition, SNP data were obtained in 50 control subjects for the GnRHR and 48 control subjects for the GPR54 gene. Statistical analysis was performed by applying a commercially available software package (GraphPad Prism; GraphPad Software, Inc., San Diego, CA, USA). Data were analyzed for normal distribution. Data are presented as means ± S.E.M.

Unpaired t-test, one-way ANOVA, Fisher’s exact test and a Chi-square test were used for the analysis of the data. P values <0.05 were considered statistically significant.


   Results
Top
 Abstract
 Introduction
 Subjects and methods
 Results
GnRHR gene
 GPR54 gene
 Discussion
 References
 
The basal hormone levels of patients and controls are shown in Table 3Go. Serum LH, FSH, testosterone and estradiol levels were significantly different between patients and controls (t-test: P < 0.001), while no significant differences were observed for SHBG.


View this table:
[in this window]
[in a new window]
  		Table 3 Serum hormone values (means ± S.E.M.) in the subjects investigated.
 

   GnRHR gene
Top
 Abstract
 Introduction
 Subjects and methods
 Results
 GnRHR gene
GPR54 gene
 Discussion
 References
 
One man with familial IHH was found to be heterozygous for a GnRHR mutation (c.785G > A) exchanging arginine for glutamine at position 262 (R262Q). No other mutation could be identified upon complete sequencing of the GnRHR gene. This patient (karyotype 46, XY) was the fourth son of a kindred of 17 siblings and was diagnosed with IHH at the age of 23 years. One of his sisters was reported to have primary amenorrhea and IHH was diagnosed. She was under hormonal treatment and adopted two children. The patient was previously treated with human chorionic gonadotropin (hCG)/human menopausal gonadotrophin (hMG), leading to an increase of testicular volume up to 11 ml and appearance of sperm in the ejaculate (maximal sperm concentration: 5 x 106/ml). He is now 53 years old and is under testosterone replacement therapy. The GPR54 gene was completely sequenced and was normal in this patient. Neither the affected sister, nor other members of his family were available for genetic screening.

No other mutations of the GnRHR gene were found in the other patients. The occurrence of a previously described, synonymous polymorphism exchanging C to T at nucleotide position 453 was investigated by restriction analysis using MaeIII (30). This polymorphism was found to be mostly heterozygous, with the same allelic frequency in both patients and controls (Table 4Go). No significant differences in the hormonal levels depending on the 453C > T SNP were observed within the groups (data not shown).


View this table:
[in this window]
[in a new window]
  		Table 4 Allelic frequency of the SNPs in the GnRHR and GPR54 genes. Complete SNP genotyping was obtained for 43 patients and either 50 (GnRHR) or 48 (GPR54) controls.
 

   GPR54 gene
Top
 Abstract
 Introduction
 Subjects and methods
 Results
 GnRHR gene
 GPR54 gene
Discussion
 References
 
In another patient with isolated IHH a homozygous sequence variant in the GPR54 gene was found (Fig. 1Go). This consisted of a homozygous insertion of a cytosine after nucleotide position 1001 (1001_1002insC). This insertion results in a frame shift in the open reading frame with elongation of the GPR54 protein from 398 to 441 amino acids.



View larger version (18K):
[in this window]
[in a new window]
  		Figure 1 Amino acid structure resulting from the homozygous 1001_1002insC sequence variant of the GPR54 gene detected in one patient with hypogonadotropic hypogonadism. The mutated sequence (mut GPR 54) is depicted below the wild-type sequence (wt GPR 54). Only the carboxyl-terminal part of the receptor, starting from amino acid 301 and including the seventh transmembrane domain (TM7) is shown. An arrow indicates the mutation site. The amino acid sequence resulting from the mutation is shown in italics.

 
The patient (karyotype 46, XY) was the only child of parents of German descent following an uneventful pregnancy. His parents were cousins of second degree. No known family history of hypogonadism or infertility was reported. He suffered from delayed puberty and bilateral undescended testes. Orchidopexy was performed at the age of 18. At the age of 20 he underwent hormonal evaluation, which showed moderately low LH (1.1 IU/l) and FSH (2.2 IU/l) levels along with reduced testosterone (3.4 nmol/l). A normal gonadotropin response to GnRH administration (100 µg i.v.) was recorded (LH peak: 10.5 IU/l; FSH peak 5.7 IU/l). He underwent testosterone enanthate replacement therapy, which induced complete virilization. At the age of 31, pulsatile GnRH therapy (5 µg/120 min s.c.) was started byan external institution in order to stimulate spermatogenesis and achieve pregnancy. The patient was referred to our Institute at the age of 32, while undergoing pulsatile GnRH treatment. He was 1.79 m tall with a body weight of 79 kg and a body mass index of 25.5 kg/m2. He was normally virilized, with mild bilateral lipomastia. Testosterone levels were in the normal range (15.6 nmol/l) while gonadotropins were increased (LH: 11.5 IU/l; FSH 24.9 IU/l). A mild hypospadia was present. Ultrasound scan showed both testes in the scrotal sac with reduced volume (right: 3 ml; left 5.3 ml). Semen analysis revealed a normal semen volume (5 ml) with oligoasthenozoospermia (concentration 0.4 x 106/ml; A + B motility 34%; 30% normal forms). At the age of 33, having been under pulsatile GnRH therapy for 2 years, he fathered a healthy son following an IVF procedure. The patient is currently 46 years old and is under hCG and hMG therapy. The patient’s parents were not available for further examination.

No other mutation was found in the patient group. Two SNPs, in exon 1 and exon 5 were found. The 24G > A variant is a synonymous SNP, while the 1091A > T polymorphism exchanges the amino acid histidine for leucine at codon 364 (H364L) in the intracellular domain. The allelic frequency of both polymorphisms was the same in patients and controls (Table 4Go). No significant differences in the hormonal levels depending on the SNPs were observed within both groups of patients and controls by ANOVA (data not shown). The combination of the two SNPs gives rise to four haplotypes, the distribution pattern of which did not differ in patients and controls (Table 5Go).


View this table:
[in this window]
[in a new window]
  		Table 5 GPR54 haplotype distribution in patients (n = 43) and controls (n = 48).
 
Considering the three SNPs in the GnRHR and GPR54 genes together, 13 combinations were observed, again with similar frequency distribution in both groups (Table 6Go). No difference in hormone concentrations was found in correlation with each combination within both groups of patients and controls by ANOVA (not shown).


View this table:
[in this window]
[in a new window]
  		Table 6 Frequency distribution of the genotype combinations resulting from the three SNPs considered in patients (n = 43) and controls (n = 48).
 

   Discussion
Top
 Abstract
 Introduction
 Subjects and methods
 Results
 GnRHR gene
 GPR54 gene
 Discussion
References
 
The pathogenesis of IHH has not yet been clarified in all cases. One possible mechanism responsible for the clinical picture of IHH is represented by mutations in the GnRHR gene. In the present study, screening for GnRHR mutations in a group of 45 hypogonadal IHH patients resulted in the detection of only one heterozygous R262Q mutation in one patient with familial IHH. From our data it is not possible to conclude that this heterozygous sequence variant plays any role in the observed phenotype. The same heterozygous mutation was reported in one patient with IHH by Beranova et al. (31) in the absence of other sequence abnormalities. However, the same heterozygous R262Q mutation was associated with normal gonadal function in other subjects (11).

The R262Q mutation in the third intracellular loop has been repeatedly described in IHH patients and, in in vitro studies, results in only partial impairment of signal transduction with decreased activation of phospholipase C without modifying ligand binding (17, 32). In our patient other sequence variants were found neither in the GnRHR nor in the GPR54 gene, leaving the pathogenesis of normosmic IHH unresolved. The heterozygous R262Q mutation might be unrelated to the IHH of our patient. Alternatively, a second mutation occurring in genomic regions not analyzed in the present study, such as the promoter or the introns, perhaps influencing gene transcription and/or splicing, might be involved. Unfortunately, other members of the family were not available for the analysis. Sixteen different GnRHR gene mutations have been described in the literature so far (4, 11, 12, 15–17, 19, 20, 22, 31, 33–36). It is worth noting that there is a certain variability in the clinical manifestation, and identical GnRHR mutations do not lead to identical phenotypes (32, 37). In accord with the literature, our data suggest that mutations of the GnRHR gene are rare in non-familial cases of IHH and mutation analysis is principally indicated in patients with normosmic autosomal recessive familial IHH.

We describe the occurrence of a novel homozygous mutation of the GPR54 gene in a male patient with IHH. Three previous studies have reported seven different mutations in five families or individuals with IHH (23, 24, 26). The sequence variant 1001_1002insC described in the present study is the first case of an insertional mutation leading to a frame shift. The insertion is located immediately 3' of the seventh transmembrane segment, in the intracellular domain of the receptor, and results in a significant alteration of the protein structure with a remarkable stretch of proline residues conferring high hydrophobicity. Although functional studies were not performed, such a drastic structural change is compatible with impairment of signal transduction, most probably resulting in the clinical phenotype. Alternatively, mRNAs containing a premature stop codon or lacking an in-frame termination codon are thought to be eliminated by ad hoc surveillance cellular mechanisms called nonsense-mediated decay and non-stop decay (38, 39) and it is therefore possible that the aberrant mRNA originating from the mutation is eliminated before transcription. In any case, an important functional impairment of the GPR54 gene in our patient should be expected. Of particular clinical interest is the fact that the patient was successfully treated with pulsatile GnRH therapy, achieving normal virilization and sperm maturation sufficient to induce a pregnancy with assisted fertilization. This is a clear, direct demonstration in the human that the KISS1/GPR54 system is operative above the GnRH/GnRHR level, possibly stimulating GnRH processing or secretion, as shown previously in rodents (40–42) and non-human primates (43).

The previously described mutations of the GPR54 gene in patients with IHH were found in a family with a homozygous 155 bp deletion at the intron 4–exon 5 junction (23), two families with homozygous missense mutations, L102P and L148S respectively (23, 24) and two isolated, non-familial cases with compound heterozygosity, i.e. R331X plus X399 (24) and C223R plus R297L (26). Altogether, our data and that of the literature now report data on 128 patients with IHH without anosmia and normal GnRHR. Although the methodology used in the different studies was different, these data suggest a higher prevalence of GPR54 mutations among familial (2 out of 13: 15.4%) than non-familial cases (3 out of 115: 2.6%). Our patient represents the first case of a homozygous sequence variant in a consanguineous family occurring as a non-familial IHH case.

Considering both genes together, according to the literature the probability of detecting mutations in either the GnRHR or the GPR54 gene in familial cases of IHH without anosmia can be as high as 50% (26, 31) so that genetic screening of these two genes is strongly indicated in such cases. The much lower incidence of the non-familial cases (<10% according to the literature, 1 case out of 39 (2.6%) detected in the present investigation) reflects the autosomal recessive nature of the disease. Unfortunately, the response to functional tests (GnRH stimulation test, monitoring of gonadotropin pulsatile secretion) in cases with mutations is rather heterogeneous and has no predictive value for identifying the carriers of mutations. The large majority of sporadic cases of IHH without anosmia remain idiopathic and other, still unrecognized factors are obviously involved in the pathogenesis of the disease. The KISS1 gene represents an obvious candidate to screen for, given its clear functional role in the GPR54–GnRHR axis (40, 41). No mutations of the KISS1 gene were found in 30 patients with IHH or constitutional delay of puberty so far (26) but more cases should be investigated in the future in order to understand the role of the putative GPR54 ligand in the activation of the hypothalamo–pituitary–gonadal axis and in the pathogenesis of IHH.

In this study we characterized SNPs in both the GnRHR and the GPR54 genes in hypogonadal and control subjects. The 453C > T synonymous SNP of the GnRHR gene showed an allele frequency of 0.080 and 0.092 in controls and patients respectively, similar to the estimated allele frequency available in the NCBI SNP database (0.091). The two other SNPs in the coding region of the GnRHR gene reported in the database, i.e. the 549G > C (rs13130501) and the 550C > G (rs13149772), were not observed in the present investigation and must have an allele frequency <0.005, since they were not found in 198 chromosomes analyzed. From our data it does not appear that common SNPs in the coding region of the GnRHR gene are associated with IHH, in agreement with a recent comprehensive study which characterized the role of GnRHR and the GnRH1 haplotypes in pubertal timing (44).

As for the GPR54 gene, the synonymous 24A > G SNP was analyzed for the first time in this study and showed a similar allele frequency in patients and controls. Similarly, no difference in the frequency distribution of the non-synonymous SNP 1091A > T was observed between the two groups, in accord with the data previously presented by Semple et al. (26). Even when the GPR54 haplotypes or the genotype combinations of both genes were considered together, no significant difference was found between patients and controls. The two additional exonic SNPs of the GPR54 gene reported in the database, i.e. the 615C > A (rs8111938) and the 155A > G (rs3746147) were not found, confirming the very low allele frequency reported by Semple et al. (26). Our data suggest that SNPs of the GnRHRand the GPR54 gene are not associated with IHH.

In conclusion, we found a novel homozygous sequence variant of the GPR54 gene in a patient with consanguineous parents and non-familial IHH. The effective stimulation of gonadotropin secretion, testosterone and spermatogenesis achieved by pulsatile GnRH therapy, showed that the KISS1/GPR54 system is necessary to stimulate GnRH secretion and/or action. The role of GPR54 in the regulation of the release of GnRH, as recently investigated in various experimental models (40, 42, 43, 45) has implications for the understanding of puberty and its disorders, as well as for possible development of ligands to manipulate the function of this axis for therapeutic purposes, e.g. suppression of the gonadal axis in patients with metastatic prostate cancer. Mutations of the GnRHR and of the GPR54 gene should be searched for in patients with IHH, especially in familial cases and when an autosomal recessive mode of inheritance is suspected.


   Acknowledgements
 
The technical assistance of Martina Niemeier and Sabine Borchert is gratefully acknowledged. We thank Susan Nieschlag for language editing of the manuscript.


   References
Top
 Abstract
 Introduction
 Subjects and methods
 Results
 GnRHR gene
 GPR54 gene
 Discussion
 References
 

    1. Seminara SB, Hayes FJ & Crowley WF. Gonadotropin-releasing hormone deficiency in the human (idiopathic hypogonadotropic hypogonadism and Kallmann’s syndrome): pathophysiological and genetic considerations. Endocrine Reviews 1998 19 521–539.[Abstract/Free Full Text]

    2. Behre HM, Nieschlag E, Meschede D & Partsch CJ. Diseases of the hypothalamus and the pituitary gland. In Andrology. Male Reproductive Health and Dysfunction, edn. 2, pp 125–142. Eds E Nieschlag & HM Behre. Berlin: Springer, 2000.

    3. Iovane A, Aumas C & de Roux N. New insights in the genetics of isolated hypogonadotropic hypogonadism. European Journal of Endocrinology 2004; 151: (Suppl 3) U83–U88.[Abstract]

    4. Silveira LF, MacColl GS & Bouloux PM. Related hypogonadotropic hypogonadism. Seminars in Reproductive Medicine 2002 20 327–338.[Web of Science][Medline]

    5. Bouloux PM, Hu Y & MacColl G. Recent advances in the pathogenesis of Kallmann’s syndrome. Progress in Brain Research 2002 141 79–83.[Web of Science][Medline]

    6. Bick D, Franco B, Sherins RJ, Heye B, Pike L, Crawford J, Maddalena A, Incerti B, Pragliola A, Meitinger T & Ballabio A. Intragenic deletion of the Kalig 1 gene in Kallmann’s syndrome. New England Journal of Medicine 1992 326 1752–1755.[Web of Science][Medline]

    7. Hardelin JP, Levilliers J, del Castillo I, Cohen-Salmon M, Legouis R, Blanchard S, Compain S, Bouloux P, Kirk J, Moraine C, Chaussain J, Weissenbach J & Petit C. X chromosome-linked Kallmann’s syndrome: stop mutations validate the candidate gene. PNAS 1992 89 8190–8194.[Abstract/Free Full Text]

    8. Zanaria E, Muscatelli F, Bardoni B, Strom TM, Guioli S, Guo W, Lalli E, Moser C, Walker AP, McCabe ER, Meitinger T, Monaco AP, Sassone-Corsi P & Camerino G. An unusual member of the nuclear hormone receptor superfamily responsible for X-linked adrenal hypoplasia congenita. Nature 1994 372 635–641.[CrossRef][Medline]

    9. Dode C, Levilliers J, Dupont JM, De Paepe A, Le Du N, Soussi-Yanicostas N, Coimbra RS, Delmaghani S, Compain-Nouaille S, Baverel F, Pecheux C, Le Tessier D, Cruaud C, Delpech M, Speleman F, Vermeulen S, Amalfitano A, Bachelot Y, Bouchard P, Cabrol S, Carel JC, Delemarre-van de Waal H, Goulet-Salmon B, Kottler ML, Richard O, Sanchez-Franco F, Saura R, Young J, Petit C & Hardelin JP. Loss-of-function mutations in FGFR1 cause autosomal dominant Kallmann syndrome. Nature Genetics 2003 33 463–465.[CrossRef][Web of Science][Medline]

    10. Sato N, Katsumata N, Kagami M, Hasegawa T, Hori N, Kawakita S, Minowada S, Shimotsuka A, Shishiba Y, Yokozawa M, Yasuda T, Nagasaki K, Hasegawa D, Hasegawa Y, Tachibana K, Naiki Y, Horikawa R, Tanaka T & Ogata T. Clinical assessment and mutation analysis of Kallmann syndrome 1 (KAL1) and fibroblast growth factor receptor 1 (FGFR1, or KAL2) in five families and 18 sporadic patients. Journal of Clinical Endocrinology and Metabolism 2004 89 1079–1088.[Abstract/Free Full Text]

    11. de Roux N, Young J, Misrahi M, Genet R, Chanson P, Schaison G & Milgrom E. A family with hypogonadotropic hypogonadism and mutations in the gonadotropin-releasing hormone receptor. New England Journal of Medicine 1997 337 1597–1602.[Free Full Text]

    12. Layman LC, Cohen DP, Jin M, Xie J, Li Z, Reindollar RH, Bolbolan S, Bick DP, Sherins RR, Duck LW, Musgrove LC, Sellers JC & Neill JD. Mutations in gonadotropin-releasing hormone receptor gene cause hypogonadotropic hypogonadism. Nature Genetics 1998 18 14–15.[CrossRef][Web of Science][Medline]

    13. Stojilkovic SS, Reinhar J & Catt KJ. Gonadotropin-releasing hormone receptor: structure and signal transduction pathways. Endocrine Reviews 1994 15 462–499.[Abstract/Free Full Text]

    14. Cheng CK & Leung PC. Molecular biology of gonadotropin-releasing hormone (GnRH)-I, GnRH-II, and their receptors in humans. Endocrine Reviews 2005 26 283–306.[Abstract/Free Full Text]

    15. Kottler ML, Lorenzo F, Bergametti F, Commercon P, Souchier C & Counis R. Subregional mapping of the human gonadotropin-releasing hormone receptor (GnRH) gene to 4q between the markers D4S392 and D4S409. Human Genetics 1995 96 477–480.[Web of Science][Medline]

    16. Caron P, Chauvin S, Christin-Maitre S, Bennet A, Lahlou N, Counis R, Bouchard P & Kottler ML. Resistance of hypogonadic patients with mutated GnRH receptor genes to pulsatile GnRH administration. Journal of Clinical Endocrinology and Metabolism 1999 84 990–996.[Abstract/Free Full Text]

    17. de Roux N, Young J, Brailly-Tabard S, Misrahi M, Milgrom E & Schaison G. The same molecular defects of the gonadotropin-releasing hormone receptor determine a variable degree of hypogonadism in affected kindred. Journal of Clinical Endocrinology and Metabolism 1999 84 567–572.[Abstract/Free Full Text]

    18. Kottler ML, Chauvin S, Lahlou N, Harris CE, Johnston CJ, Lagarde JP, Bouchard P, Farid NR & Counis R. A new compound heterozygous mutation of the gonadotropin-releasing hormone receptor (L314X, Q106R) in a woman with complete hypogonadotropic hypogonadism: chronic estrogen administration amplifies the gonadotropin defect. Journal of Clinical Endocrinology and Metabolism 2000 85 3002–3008.[Abstract/Free Full Text]

    19. Pralong FP, Gomez F, Castillo E, Cotecchia S, Abuin L, Aubert ML & Portmann L. Complete hypogonadotropic hypogonadism associated with a novel inactivating mutation of the gonadotropin-releasing hormone receptor. Journal of Clinical Endocrinology and Metabolism 1999 84 3811–3816.[Abstract/Free Full Text]

    20. Pitteloud N, Boepple PA, DeCruz S, Valkenburgh SB, Crowley WF Jr & Hayes FJ. The fertile eunuch variant of idiopathic hypogonadotropic hypogonadism: spontaneous reversal associated with a homozygous mutation in the gonadotropin-releasing hormone receptor. Journal of Clinical Endocrinology and Metabolism 2001 86 2470–2475.[Abstract/Free Full Text]

    21. Silveira LF, Stewart PM, Thomas M, Clark DA, Bouloux PM & MacColl GS. Novel homozygous splice acceptor site GnRH receptor (GnRHR) mutation: human GnRHR ‘knockout’. Journal of Clinical Endocrinology and Metabolism 2002 87 2973–2977.[Abstract/Free Full Text]

    22. Wolczynski S, Laudanski P, Jarzabek K, Mittre H, Lagarde JP & Kottler ML. A case of complete hypogonadotropic hypogonadism with a mutation in the gonadotropin-releasing hormone receptor gene. Fertility and Sterility 2003 79 442–444.[Web of Science][Medline]

    23. de Roux N, Genin E, Carel JC, Matsuda F, Chaussain JL & Milgrom E. Hypogonadotropic hypogonadism due to loss of function of the KiSS1-derived peptide receptor GPR54. PNAS 2003 100 10972–10976.[Abstract/Free Full Text]

    24. Seminara SB, Messager S, Chatzidaki EE, Thresher RR, Acierno JS Jr, Shagoury JK, Bo-Abbas Y, Kuohung W, Schwinof KM, Hendrick AG, Zahn D, Dixon J, Kaiser UB, Slaugenhaupt SA, Gusella JF, O’Rahilly S, Carlton MB, Crowley WF Jr, Aparicio SA & Colledge WH. The GPR54 gene as a regulator of puberty. New England Journal of Medicine 2003 349 1614–1627.[Abstract/Free Full Text]

    25. Colledge WH. GPR54 and puberty. Trends in Endocrinology and Metabolism 2004 15 448–453.[CrossRef][Web of Science][Medline]

    26. Semple RK, Achermann JC, Ellery J, Farooqi IS, Karet FE, Stanhope RG, O’Rahilly S & Aparicio SA. Two novel missense mutations in G protein-coupled receptor 54 in a patient with hypogonadotropic hypogonadism. Journal of Clinical Endocrinology and Metabolism 2005 90 1849–1855.[Abstract/Free Full Text]

    27. Simoni M, Gromoll J, Höppner W, Kamischke A, Krafft T, Stähle D & Nieschlag E. Mutational analysis of the follicle-stimulating hormone (FSH) receptor in normal and infertile men: identification and characterization of two discrete FSH receptor isoforms. Journal of Clinical Endocrinology and Metabolism 1999 84 751–755.[Abstract/Free Full Text]

    28. Gromoll J, Brocker M, Derwahl M & Hoppner W. Detection of mutations in glycoprotein hormone receptors. Methods 2000 21 83–97.[CrossRef][Web of Science][Medline]

    29. von Eckardstein S, Schmidt A, Kamischke A, Simoni M, Gromoll J & Nieschlag E. CAG repeat length in the androgen receptor gene and gonadotrophin suppression influence the effectiveness of hormonal male contraception. Clinical Endocrinology 2002 57 647–655.[CrossRef][Medline]

    30. Kottler ML, Seret-Begue D, Lahlou N, Assayag M, Carre MC, Lagarde JP, Ajzenberg C, Christin-Maitre S, Bouchard P, Mikol J, Counis R & Warnet A. The GnRH receptor gene is preferentially expressed in functioning gonadotroph adenomas and displays a MaeIII polymorphism site. Clinical Endocrinology 1998 49115–123.[CrossRef][Medline]

    31. Beranova M, Oliveira LMB, Bédécarrats GY, Schipani E, Vallejo M, Ammini AC, Quintos JB, Hall JE, Martin KA, Hayes FJ, Pitteloud N, Kaiser UB, Crowley WF Jr & Seminara SB. Prevalence, phenotypic spectrum, and modes of inheritance of gonadotropin-releasing hormone receptor mutations in idiopathic hypogonadotropic hypogonadism. Journal of Clinical Endocrinology and Metabolism 2001 86 1580–1588.[Abstract/Free Full Text]

    32. Karges B, Karges W & de Roux N. Clinical and molecular genetics of the human GnRH receptor. Human Reproduction Update 2003 9 523–530.[Abstract/Free Full Text]

    33. Seminara SB, Oliveira LM, Beranova M, Hayes FJ & Crowley WF Jr. Genetics of hypogonadotropic hypogonadism. Journal of Endocrinological Investigation 2000 23 560–565.[Web of Science][Medline]

    34. Soderlund D, Canto P, de la Chesnaye E, Ulloa-Aguirre A & Mendez JP. A novel homozygous mutation in the second transmembrane domain of the gonadotrophin releasing hormone receptor gene. Clinical Endocrinology 2001 54 493–498.[CrossRef][Medline]

    35. Costa EM, Bedecarrats GY, Mendonca BB, Arnhold IJ, Kaiser UB & Latronico AC. Two novel mutations in the gonadotropin-releasing hormone receptor gene in Brazilian patients with hypogonadotropic hypogonadism and normal olfaction. Journal of Clinical Endocrinology and Metabolism 2001 86 2680–2686.[Abstract/Free Full Text]

    36. Karges B, Karges W, Mine M, Ludwig L, Kuhne R, Milgrom E & de Roux N. Mutation Ala(171)Thr stabilizes the gonadotropin-releasing hormone receptor in its inactive conformation, causing familial hypogonadotropic hypogonadism. Journal of Clinical Endocrinology and Metabolism 2003 88 1873–1879.[Abstract/Free Full Text]

    37. Karges B & de Roux N. Molecular genetics of isolated hypogonadotropic hypogonadism and Kallmann syndrome. Endocrine Development 2005 8 67–80.[Medline]

    38. Wagner E & Lykke-Andersen J. mRNA surveillance: the perfect persist. Journal of Cell Science 2002 115 3033–3038.[Abstract/Free Full Text]

    39. Chatr-Aryamontri A, Angelini M, Garelli E, Tchernia G, Ramenghi U, Dianzani I & Loreni F. Nonsense-mediated and nonstop decay of ribosomal protein S19 mRNA in Diamond-Black-fan anemia. Human Mutation 2004 24 526–533.[CrossRef][Web of Science][Medline]

    40. Irwig MS, Fraley GS, Smith JT, Acohido BV, Popa SM, Cunningham MJ, Gottsch ML, Clifton DK & Steiner RA. Kisspeptin activation of gonadotropin releasing hormone neurons and regulation of KiSS-1 mRNA in the male rat. Neuroendocrinology 2004 80 264–272.[CrossRef][Web of Science][Medline]

    41. Brailoiu GC, Dun SL, Ohsawa M, Yin D, Yang J, Chang JK, Brailoiu E & Dun NJ. KiSS-1 expression and metastin-like immunoreactivity in the rat brain. Journal of Comparative Neurology 2005 481 314–329.[CrossRef][Web of Science][Medline]

    42. Messager S, Chatzidaki EE, Ma D, Hendrick AG, Zahn D, Dixon J, Thresher RR, Malinge I, Lomet D, Carlton MB, Colledge WH, Caraty A & Aparicio SA. Kisspeptin directly stimulates gonadotropin-releasing hormone release via G protein-coupled receptor 54. PNAS 2005 102 1761–1766.[Abstract/Free Full Text]

    43. Shahab M, Mastronardi C, Seminara SB, Crowley WF, Ojeda SR & Plant TM. Increased hypothalamic GPR54 signaling: a potential mechanism for initiation of puberty in primates. PNAS 2005 102 2129–2134.[Abstract/Free Full Text]

    44. Sedlmeyer IL, Pearce CL, Trueman JA, Butler JL, Bersaglieri T, Read AP, Clayton PE, Kolonel LN, Henderson BE, Hirschhorn JN & Palmert MR. Determination of sequence variation and haplo-type structure for the gonadotropin-releasing hormone (GnRH) and GnRH receptor genes: investigation of role in pubertal timing. Journal of Clinical Endocrinology and Metabolism 2005 90 1091–1099.[Abstract/Free Full Text]

    45. Navarro VM, Castellano JM, Fernandez-Fernandez R, Tovar S, Roa J, Mayen A, Barreiro ML, Casanueva FF, Aguilar E, Dieguez C, Pinilla L & Tena-Sempere M. Effects of KiSS-1 peptide, the natural ligand of GPR54, on follicle-stimulating hormone secretion in the rat. Endocrinology 2005 146 1689–1697.[Abstract/Free Full Text]

Received 1 June 2005
Accepted 1 September 2005




This article has been cited by other articles:


Home page
 ReproductionHome page
R. M Reynolds, J. J Logie, A. K Roseweir, A. J McKnight, and R. P Millar
A role for kisspeptins in pregnancy: facts and speculations
Reproduction, July 1, 2009; 138(1): 1 - 7.
[Abstract] [Full Text] [PDF]

Home page
 Hum Reprod UpdateHome page
A.K. Roseweir and R.P. Millar
The role of kisspeptin in the control of gonadotrophin secretion
Hum. Reprod. Update, March 1, 2009; 15(2): 203 - 212.
[Abstract] [Full Text] [PDF]

Home page
 Biol. Reprod.Home page
A. S. Mechaly, J. Vinas, and F. Piferrer
Identification of Two Isoforms of the Kisspeptin-1 Receptor (kiss1r) Generated by Alternative Splicing in a Modern Teleost, the Senegalese Sole (Solea senegalensis)
Biol Reprod, January 1, 2009; 80(1): 60 - 69.
[Abstract] [Full Text] [PDF]

Home page
 ReproductionHome page
E. Gianetti and S. Seminara
Kisspeptin and KISS1R: a critical pathway in the reproductive system
Reproduction, September 1, 2008; 136(3): 295 - 301.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
X. d'Anglemont de Tassigny, L. A. Fagg, J. P. C. Dixon, K. Day, H. G. Leitch, A. G. Hendrick, D. Zahn, I. Franceschini, A. Caraty, M. B. L. Carlton, et al.
Hypogonadotropic hypogonadism in mice lacking a functional Kiss1 gene
PNAS, June 19, 2007; 104(25): 10714 - 10719.
[Abstract] [Full Text] [PDF]

Home page
 J. Clin. Endocrinol. Metab.Home page
Y. Tenenbaum-Rakover, M. Commenges-Ducos, A. Iovane, C. Aumas, O. Admoni, and N. de Roux
Neuroendocrine Phenotype Analysis in Five Patients with Isolated Hypogonadotropic Hypogonadism due to a L102P Inactivating Mutation of GPR54
J. Clin. Endocrinol. Metab., March 1, 2007; 92(3): 1137 - 1144.
[Abstract] [Full Text] [PDF]

Home page
 Eur J EndocrinolHome page
F. Cerrato, J. Shagoury, M. Kralickova, A. Dwyer, J. Falardeau, M. Ozata, G. Van Vliet, P. Bouloux, J. E Hall, F. J Hayes, et al.
Coding sequence analysis of GNRHR and GPR54 in patients with congenital and adult-onset forms of hypogonadotropic hypogonadism
Eur. J. Endocrinol., November 1, 2006; 155(suppl_1): S3 - S10.
[Abstract] [Full Text] [PDF]

Home page
 Hum Reprod UpdateHome page
M. Tena-Sempere
GPR54 and kisspeptin in reproduction
Hum. Reprod. Update, September 1, 2006; 12(5): 631 - 639.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via ISI Web of Science (20)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Lanfranco, F.
Right arrow Articles by Simoni, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Lanfranco, F.
Right arrow Articles by Simoni, M.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS