Section of Reproductive Endocrinology, Infertility, and Genetics, Department of Obstetrics and Gynecology, Medical College of Georgia, Augusta, Georgia 30912, USA.
Fertil Steril. 2011 Apr;95(5):1613-20.e1-7. doi: 10.1016/j.fertnstert.2011.01.010. Epub 2011 Feb 15.
To determine if mutations in NELF, a gene isolated from migratory GnRH neurons, cause normosmic idiopathic hypogonadotropic hypogonadism (IHH) and Kallmann syndrome (KS).
Molecular analysis correlated with phenotype.
Academic medical center.
PATIENT(S): A total of 168 IHH/KS patients as well as unrelated control subjects were studied for NELF mutations.
INTERVENTION(S): NELF coding regions/splice junctions were subjected to polymerase chain reaction (PCR)-based DNA sequencing. Eleven additional IHH/KS genes were sequenced in three patients with NELF mutations.
MAIN OUTCOME MEASURE(S): Mutations were confirmed by sorting intolerant from tolerant, reverse-transcription (RT)-PCR, and Western blot analysis.
RESULT(S): Three novel NELF mutations absent in 372 ethnically matched control subjects were identified in 3/168 (1.8%) IHH/KS patients. One IHH patient had compound heterozygous NELF mutations (c.629-21G>C and c.629-23C>G), and he did not have mutations in 11 other known IHH/KS genes. Two unrelated KS patients had heterozygous NELF mutations and mutation in a second gene: NELF/KAL1 (c.757G>A; p.Ala253Thr of NELF and c.488_490delGTT; p.Cys163del of KAL1) and NELF/TACR3 (c.1160-13C>T of NELF and c.824G>A; p.Trp275X of TACR3). In vitro evidence of these NELF mutations included reduced protein expression and splicing defects.
CONCLUSION(S): Our findings suggest that NELF is associated with normosmic IHH and KS, either singly or in combination with a mutation in another gene.
确定从迁徙性 GnRH 神经元中分离出的 NELF 基因突变是否会导致正常嗅觉性特发性促性腺激素低下性性腺功能减退症 (IHH) 和卡尔曼综合征 (KS)。
表型相关的分子分析。
学术医疗中心。
总共研究了 168 名 IHH/KS 患者和无关的对照，以寻找 NELF 突变。
对 NELF 编码区/剪接接头进行基于聚合酶链反应 (PCR) 的 DNA 测序。在 3 名有 NELF 突变的患者中，还对另外 11 个 IHH/KS 基因进行了测序。
通过排序不耐受与耐受、逆转录 (RT)-PCR 和 Western blot 分析来确认突变。
在 168 名 IHH/KS 患者中，有 3 名患者 (1.8%) 发现了 3 个新的 NELF 突变，这些突变不存在于 372 名种族匹配的对照中。1 名 IHH 患者存在复合杂合 NELF 突变 (c.629-21G>C 和 c.629-23C>G)，且他没有其他已知的 IHH/KS 基因的突变。2 名无关的 KS 患者存在杂合 NELF 突变和第二个基因的突变：NELF/KAL1 (c.757G>A;p.Ala253Thr 于 NELF 和 c.488_490delGTT;p.Cys163del 于 KAL1) 和 NELF/TACR3 (c.1160-13C>T 于 NELF 和 c.824G>A;p.Trp275X 于 TACR3)。这些 NELF 突变的体外证据包括蛋白表达减少和剪接缺陷。
我们的研究结果表明，NELF 与正常嗅觉性 IHH 和 KS 相关，无论是单独发生还是与另一个基因突变组合发生。
