Purpose

To screen novel mutations in LHCGR responsible for empty follicle syndrome and explore the pathological mechanism of mutations.

Methods

Four affected individuals diagnosed with infertility-associated anovulation or oligo-ovulation from three independent families were recruited. Sanger sequencing was used to identify the LHCGR mutations in affected individuals. Western blot was performed to evaluate the effects of mutations on LHCGR protein levels. Immunofluorescence was done to explore the effects of mutations on LHCGR subcellular localization. The ATP levels were measured to infer the functional effects of the mutations on LHCGR.

Results

In the present study, three novel biallelic mutations in LHCGR were identified in four affected individuals from three independent families with empty follicle syndrome or oligo-ovulation. All biallelic mutations were inherited from the proband of their parents. The western blot showed that the identified mutations decreased LHCGR protein level and altered the glycosylation pattern. The immunofluorescence showed an ectopic subcellular localization of LHCGR in cultured HeLa cells. Besides, the mutations in LHCGR also reduced the cellular ATP consumption.

Conclusion

These findings confirm previous studies and expand the mutational spectrum of LHCGR, which will provide genetic diagnostic marker for patients with empty follicle syndrome.

中文翻译：

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LHCGR（促黄体激素/绒毛膜促性腺激素受体）的新突变：扩大导致人类空卵泡综合征的突变谱。

目的

筛选导致空卵泡综合征的LHCGR新突变并探索突变的病理机制。

方法

招募了来自三个独立家庭的四名被诊断患有与不孕症相关的无排卵或少排卵的受影响个体。桑格测序用于鉴定受影响个体的LHCGR突变。进行蛋白质印迹以评估突变对 LHCGR 蛋白水平的影响。进行免疫荧光以探索突变对 LHCGR 亚细胞定位的影响。测量 ATP 水平以推断突变对 LHCGR 的功能影响。

结果

在本研究中，在来自具有空卵泡综合征或少排卵的三个独立家庭的四个受影响个体中发现了LHCGR中的三个新的双等位基因突变。所有双等位基因突变均遗传自其父母的先证者。蛋白质印迹表明，鉴定的突变降低了 LHCGR 蛋白水平并改变了糖基化模式。免疫荧光显示 LHCGR 在培养的 HeLa 细胞中异位亚细胞定位。此外，LHCGR的突变也减少了细胞 ATP 的消耗。

结论

这些发现证实了之前的研究并扩大了LHCGR的突变谱，这将为空卵泡综合征患者提供遗传诊断标志物。