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Two Novel Mutations in the Gonadotropin-Releasing Hormone Receptor Gene in Brazilian Patients with Hypogonadotropic Hypogonadism and Normal Olfaction¹

Unidade de Endocrinologia do Desenvolvimento e Laboratório de Hormônios e Genética Molecular LIM/42, da Disciplina de Endocrinologia do Hospital das Clínicas da Faculdade de Medicina da Universidade de Sao Paulo (E.M.F.C., B.B.M., I.J.P.A., A.C.L.), Sao Paulo 01060–970, Brazil; and Endocrine Hypertension Division, Department of Medicine, Brigham and Women’s Hospital and Harvard Medical School (U.B.K., G.Y.B.), Boston, Massachusetts 02115

Address all correspondence and requests for reprints to: Dr. Ana Claudia Latronico, Hospital das Clínicas, Faculdade de Medicina da Universidade de Sao Paulo, Disciplina de Endocrinologia, Universidade de Sao Paulo. Caixa Postal 3671, Sao Paulo 01060-970, Brazil. E-mail: anacl{at}usp.br

Abstract

Several point mutations in the GnRH receptor gene have been described in an autosomal recessive form of congenital isolated hypogonadotropic hypogonadism (HH). We investigated 17 Brazilian patients (10 males and 7 females) from 14 different families, with HH and normal olfaction. The diagnosis of HH was based on absent or incomplete sexual development after 17 yr of age associated with low or normal levels of LH in both sexes and low levels of testosterone in males and of estradiol in females. All patients presented with a normal sense of smell in an olfactory specific test.

The coding region of the GnRH receptor gene was amplified by PCR and directly sequenced. A novel missense mutation, Arg¹³⁹His, located in the conserved DRS motif at the junction of the third transmembrane and the second intracellular loop of the GnRH receptor was identified in the homozygous state in one female with complete HH. The Arg¹³⁹His mutation completely eliminated detectable GnRH-binding activity and prevented GnRH-induced stimulation of inositol phosphate accumulation in vitro.

In another family, a new compound heterozygous mutation (Asn¹⁰Lys and Gln¹⁰⁶Arg) was identified in four siblings (two males and two females) with partial HH. The Gln¹⁰⁶Arg mutation, located in the first extracellular loop, has been previously described, and in vitro analysis indicated that the mutant receptor was able to bind GnRH, but with a reduced affinity. The Asn¹⁰Lys mutation in the extracellular amino-terminal domain of the receptor also reduced the affinity for GnRH in vitro. In this family we also identified a previously described silent polymorphism at amino acid residue 151 in the second intracellular loop that segregated with the two inactivating mutations of the GnRH receptor. This polymorphism was also found in two unrelated patients with sporadic HH without GnRH receptor loss of function mutations. No mutations were identified in the remaining cases.

A good correlation between genotype and phenotype was found in our patients. The woman, who is homozygous for the completely inactivating Arg¹³⁹His mutation, has complete HH with undetectable serum basal LH and FSH levels that failed to respond to GnRH stimulation. In addition, the affected patients who are compound heterozygotes for the Asn¹⁰Lys/Gln¹⁰⁶Arg mutations, have partial HH with low serum basal LH levels that were responsive to GnRH stimulation.

No clinical or hormonal differences were found between HH patients with and without mutations in the GnRH receptor gene, indicating that these data do not contribute to the identification of HH patients with GnRH receptor mutations. In conclusion, we report the first naturally occurring mutation within the conserved DRS motif of the GnRH receptor in a female with complete HH and a novel compound heterozygous mutation (Asn¹⁰Lys and Gln¹⁰⁶Arg) in a family with partial HH, increasing the repertoire of the inactivating mutations of the GnRH receptor.

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