精子鞭毛多发形态异常患者KLHL10基因突变分析
Detection of KLHL10 gene mutations in patients with multiple morphological abnormalities of the sperm flagella
张慧 1黄吴键 2陈国勇 2梅晓妍 2谢鹏 2郑海英 3兰风华 4张朵4
作者信息
- 1. 中国人民解放军联勤保障部队第九○○医院基础医学实验室,福建福州 350025;福建省移植生物学重点实验室,福建福州 350025
- 2. 中国人民解放军联勤保障部队第九○○医院生殖医学中心,福建福州 350025
- 3. 中国人民解放军联勤保障部队第九○○医院基础医学实验室,福建福州 350025;福建医科大学福总临床医学院,福建福州 350108
- 4. 中国人民解放军联勤保障部队第九○○医院基础医学实验室,福建福州 350025;福建省移植生物学重点实验室,福建福州 350025;福建医科大学福总临床医学院,福建福州 350108
- 折叠
摘要
目的 探寻精子鞭毛多发形态异常患者的致病基因.方法 通过全外显子组测序(WES)对患者样本进行检测,分析可能的致病基因;运用扫描电镜和透射电镜观察患者精子超微结构;通过定量聚合酶链反应(qPCR)分析精子中目的基因mRNA表达水平;通过PubMed、中国知识基础设施工程(CNKI)等数据库检索目的基因突变相关临床病例.结果 在本研究征集的患者中检测到一个KLHL10基因杂合突变:c.317C＞T(p.Pro106Leu),透射电镜结果表明患者精子鞭毛结构缺失或紊乱,未能找见轴丝中典型"9+2"结构.qPCR结果提示患者精子KLHL10 mRNA表达水平降低,只有正常对照组的一半,而与KLHL10相互结合的泛素连接酶CUL3(Cullin-3)的mRNA表达量却较正常对照升高2.5倍.结论 本研究报道了精子鞭毛多发形态异常患者KLHL10基因C.317C＞T突变,该突变位点首次报道可能通过单倍体剂量不足引起精子形态异常.
Abstract
Objective Exploring the possible pathogenic genes in patients with multiple morphological abnormalities of the sperm flagella(MMAF).Methods The samples of MMAF patients were detected by whole exome sequencing(WES)to analyze possible pathogenic genes.Using scanning electron microscopy and transmission electron microscopy to observe the ultrastructure of sperm in MMAF patients.Analyze the mRNA expression of the target gene in the patient's sperm through qPCR.Results A heterozygous mutation in the KLHL10 gene was detected in one MMAF patient:c.317C＞T(p.Pro106Leu).Transmission electron microscopy results showed that the ultrastructure of the patient's sperm flagella was missing or disor-dered,the typical"9+2"structures in the axoneme could not be found competely.The qPCR results indicated that the mRNA expression of KLHL 10 in the patient's sperm decreased,which was only half of the control.However,the expression of CUL3 which binds to KLHL10,was 2.5 times higher than the control.Conclusion This study reported a case of KLHL10 gene C.317C＞T mutation in MMAF patients,which was first reported to cause male MMAF phenotype through insufficient haploid dose.
关键词
精子鞭毛多发形态异常/全外显子组测序/KLHL10基因/男性不育Key words
MMAF/whole exome sequencing/KLHL10 gene/male infertility引用本文复制引用
基金项目
福建省自然科学基金(2020J011149)
军委后勤保障部计划生育专项课题基金(21JSZ12)
出版年
2024
NETL
NSTL
万方数据