{"title":"IQCN 的功能缺失突变会导致人类和小鼠完全受精失败,从而导致雄性不育。","authors":"Yulin Wang, Guoyong Chen, Zhenzhen Tang, Xiaoyan Mei, Chunli Lin, Jingyi Kang, Jianqing Lian, Jun Lu, Yun Liu, Fenghua Lan, Wujian Huang, Duo Zhang","doi":"10.1093/molehr/gaad018","DOIUrl":null,"url":null,"abstract":"<p><p>Fertilization failure is a significant manifestation of unexplained male infertility. Previous work has suggested a genetic origin. In this study, we report on a man with unexplained infertility from a large consanguineous marriage family. Whole-exome sequencing and Sanger sequencing identified a homozygous frameshift variation of the IQ motif containing N (IQCN; GenBank: NM_001145304.1; c.1061_1062delAT; p.Y354Sfs*13) in the proband and one of his two brothers, who also remained infertile. Analyses of spermatozoa by quantitative RT-PCR indicated that the level of IQCN mRNA was significantly reduced compared to fertile men and the protein could not be detected by western blotting and immunofluorescent staining in the proband. Immunofluorescent staining of spermatozoa from fertile men showed that IQCN was located in the acrosomal region and translocated to the equatorial segment after the acrosome reaction. The proband spermatozoa had abnormal morphology and function. Finally, the proband couple underwent IVF with donor sperm and a healthy baby was born. Furthermore, we developed an Iqcn-KO mouse model using the CRISPR/Cas9 technique. Sperm quality, except for sperm motility, and the fertility of male Iqcn-/- mice were consistent with those of the proband. In conclusion, the findings in humans and mice demonstrate that the homozygous frameshift variant of IQCN causes male infertility owing to autosomal-recessive fertilization failure.</p>","PeriodicalId":18759,"journal":{"name":"Molecular human reproduction","volume":"29 7","pages":""},"PeriodicalIF":3.6000,"publicationDate":"2023-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Loss-of-function mutations in IQCN cause male infertility in humans and mice owing to total fertilization failure.\",\"authors\":\"Yulin Wang, Guoyong Chen, Zhenzhen Tang, Xiaoyan Mei, Chunli Lin, Jingyi Kang, Jianqing Lian, Jun Lu, Yun Liu, Fenghua Lan, Wujian Huang, Duo Zhang\",\"doi\":\"10.1093/molehr/gaad018\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Fertilization failure is a significant manifestation of unexplained male infertility. Previous work has suggested a genetic origin. In this study, we report on a man with unexplained infertility from a large consanguineous marriage family. Whole-exome sequencing and Sanger sequencing identified a homozygous frameshift variation of the IQ motif containing N (IQCN; GenBank: NM_001145304.1; c.1061_1062delAT; p.Y354Sfs*13) in the proband and one of his two brothers, who also remained infertile. Analyses of spermatozoa by quantitative RT-PCR indicated that the level of IQCN mRNA was significantly reduced compared to fertile men and the protein could not be detected by western blotting and immunofluorescent staining in the proband. Immunofluorescent staining of spermatozoa from fertile men showed that IQCN was located in the acrosomal region and translocated to the equatorial segment after the acrosome reaction. The proband spermatozoa had abnormal morphology and function. Finally, the proband couple underwent IVF with donor sperm and a healthy baby was born. Furthermore, we developed an Iqcn-KO mouse model using the CRISPR/Cas9 technique. Sperm quality, except for sperm motility, and the fertility of male Iqcn-/- mice were consistent with those of the proband. In conclusion, the findings in humans and mice demonstrate that the homozygous frameshift variant of IQCN causes male infertility owing to autosomal-recessive fertilization failure.</p>\",\"PeriodicalId\":18759,\"journal\":{\"name\":\"Molecular human reproduction\",\"volume\":\"29 7\",\"pages\":\"\"},\"PeriodicalIF\":3.6000,\"publicationDate\":\"2023-06-30\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Molecular human reproduction\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1093/molehr/gaad018\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"DEVELOPMENTAL BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Molecular human reproduction","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1093/molehr/gaad018","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"DEVELOPMENTAL BIOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
受精失败是原因不明的男性不育症的一个重要表现。以往的研究表明这与遗传有关。在本研究中,我们报告了一名来自近亲结婚大家庭的男性不明原因不育症患者。通过全外显子组测序和桑格测序发现,该男性和他的两个兄弟中的一个存在含有 N 的 IQ 标记(IQCN;GenBank:NM_001145304.1;c.1061_1062delAT;p.Y354Sfs*13)的同基因框移位变异,而这两个兄弟也仍然不育。通过定量 RT-PCR 对精子进行的分析表明,IQCN mRNA 的水平与可育男性相比明显降低,而且在该受试者体内,通过免疫印迹和免疫荧光染色无法检测到该蛋白。对可育男性精子的免疫荧光染色显示,IQCN位于顶体区域,并在顶体反应后转位至赤道段。该疑似患者的精子在形态和功能上均出现异常。最后,这对疑似夫妇接受了捐赠精子的体外受精,生下了一个健康的婴儿。此外,我们还利用 CRISPR/Cas9 技术开发了 Iqcn-KO 小鼠模型。除精子活力外,雄性Iqcn-/-小鼠的精子质量和生育能力与疑似患者一致。总之,对人类和小鼠的研究结果表明,IQCN的同基因框移变体会导致常染色体隐性受精失败,从而导致男性不育。
Loss-of-function mutations in IQCN cause male infertility in humans and mice owing to total fertilization failure.
Fertilization failure is a significant manifestation of unexplained male infertility. Previous work has suggested a genetic origin. In this study, we report on a man with unexplained infertility from a large consanguineous marriage family. Whole-exome sequencing and Sanger sequencing identified a homozygous frameshift variation of the IQ motif containing N (IQCN; GenBank: NM_001145304.1; c.1061_1062delAT; p.Y354Sfs*13) in the proband and one of his two brothers, who also remained infertile. Analyses of spermatozoa by quantitative RT-PCR indicated that the level of IQCN mRNA was significantly reduced compared to fertile men and the protein could not be detected by western blotting and immunofluorescent staining in the proband. Immunofluorescent staining of spermatozoa from fertile men showed that IQCN was located in the acrosomal region and translocated to the equatorial segment after the acrosome reaction. The proband spermatozoa had abnormal morphology and function. Finally, the proband couple underwent IVF with donor sperm and a healthy baby was born. Furthermore, we developed an Iqcn-KO mouse model using the CRISPR/Cas9 technique. Sperm quality, except for sperm motility, and the fertility of male Iqcn-/- mice were consistent with those of the proband. In conclusion, the findings in humans and mice demonstrate that the homozygous frameshift variant of IQCN causes male infertility owing to autosomal-recessive fertilization failure.
期刊介绍:
MHR publishes original research reports, commentaries and reviews on topics in the basic science of reproduction, including: reproductive tract physiology and pathology; gonad function and gametogenesis; fertilization; embryo development; implantation; and pregnancy and parturition. Irrespective of the study subject, research papers should have a mechanistic aspect.