[1]李中泰,李彦锋,刘晋祎,等.Fscb/Cabyr双基因敲除雄性小鼠的精子形态学表型特征及运动参数变化[J].第三军医大学学报,2020,42(24):2358-2365.
 LI Zhongta,LI Yanfeng,LIU Jinyi,et al.Morphological phenotypes and changes in motility parameters of sperms in Fscb/Cabyr double knockout mice[J].J Third Mil Med Univ,2020,42(24):2358-2365.
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Fscb/Cabyr双基因敲除雄性小鼠的精子形态学表型特征及运动参数变化(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
42卷
期数:
2020年第24期
页码:
2358-2365
栏目:
基础医学
出版日期:
2020-12-30

文章信息/Info

Title:
Morphological phenotypes and changes in motility parameters of sperms in Fscb/Cabyr double knockout mice
作者:
李中泰李彦锋刘晋祎张勇张军毕罡
陆军军医大学(第三军医大学)大坪医院泌尿外科;陆军军医大学(第三军医大学)预防医学院毒理学研究所;中国人民解放军第990医院泌尿外科
 
Author(s):
LI Zhongta LI Yanfeng1 LIU Jinyi ZHANG Yong ZHANG Jun BI Gang

Department of Urology, Daping Hospital, Army Medical University (Third Military Medical University), Chongqing, 400042; 2Institute of Toxicology, Faculty of Military Preventive Medicine, Army Medical University (Third Military Medical University), Chongqing, 400038; 3Department of Urology, NO. 990 Hospital of PLA, Zhumadian, Henan Province, 463000, China

关键词:
 
Keywords:
gene knockout sperm fibrous sheath morphology motility
分类号:
R322.1; R329.24; R394-33
文献标志码:
A
摘要:

目的探讨Fscb/Cabyr双基因敲除后雄性小鼠精子鞭毛的形态学特征改变及精子运动功能变化情况。方法取不同基因型雄性小鼠睾丸及附睾组织,计算脏体比,睾丸组织行常规HE染色,观察其生精功能状态变化;取不同基因型雄性小鼠附睾精子,应用计算机辅助精液分析系统进行精子运动功能相关参数分析,通过体外培养观察精子获能和顶体反应情况改变,应用光镜及电镜对精子的显微形态学及超微结构进行观察。结果Fscb/Cabyr(++/+-)、Fscb/Cabyr(--/+-)、Fscb/Cabyr(++/++)、Fscb/Cabyr(--/--)、Fscb/Cabyr(++/--)等5种基因型雄性小鼠附睾精子密度差异均无统计学意义(P>0.05)。但Fscb/Cabyr(--/--)、Fscb/Cabyr(++/--)基因型雄性小鼠精子的前向运动精子比率(progressive rate,PR)和运动精子比率(PR+NP)、平均曲线运动速度(curvilinear velocity,VCL)、平均直线运动速度(straight-line velocity,VSL)、平均路径运动速度(average path velocity,VAP)、精子平均鞭打频率(beat cross frequency,BCF)和平均头部侧摆幅值(amplitude of lateral head displacement,ALH)均较野生型雄性小鼠显著降低(P<0.01)。Fscb/Cabyr(--/--)和Fscb/Cabyr(++/--)雄性小鼠正常精子百分率[(49.61±1.45)%和(53.22±3.51%)]显著低于野生型小鼠[(66.46±0.73)%,P<0.01];其精子尾折叠比例[(20.96±0.43)%和(20.73±0.21)%]显著高于野生型小鼠[(5.74±0.72)%,P<0.01]。Fscb/Cabyr(--/--)、Fscb/Cabyr(++/--)两种基因型雄性小鼠精子鞭毛的超微结构观察显示:大部分精子纤维鞘纵行柱状体结构消失,环形肋板结构紊乱,电子密度不均匀降低,呈显著疏松或增厚表现。部分精子鞭毛的双联微管结构排列不规则,存在个别缺失和中心两个微管的分离现象。结论Cabyr单基因和Fscb/Cabyr双基因敲除雄性小鼠均出现显著精子鞭毛形态学异常和运动功能降低。CABYR蛋白对纤维鞘的正确形成和精子鞭毛维持正常的运动功能是必不可少的。

Abstract:
ObjectiveTo investigate the changes in morphological characteristics of sperm flagellum and in sperm motility parameters in Fscb/Cabyr double gene knockout (Fscb/Cabyr--/--) mice. MethodsThe testis and epididymis tissues of male mice with different genotypes were collected and the organ-to-body weight ratio was calculated. The changes of spermatogenesis function in the testicular tissues of the mice were observed using HE staining. The parameters of sperm motility were analyzed using a computer-aided sperm analysis (CASA) system, and sperm capacitation and acrosome reaction were observed with in vitro culture. The morphological characteristics of the sperms under optical microscope and electron microscope were compared among the mice with different genotypes. ResultsNo significant difference was found in sperm density in the epididymis among the mice with 5 different genotypes. The progressive rate (PR) and motility rate (PR+NP), mean curvilinear velocity (VCL), mean straight-line velocity (VSL), mean path velocity (VAP), beat cross frequency (BCF) and amplitude of lateral head displacement (ALH) of the sperms were significantly lower in Fscb/Cabyr--/-- and Fscb/Cabyr++/-- mice than in the wild-type mice (P<0.01). The Fscb/Cabyr--/-- and Fscb/Cabyr++/-- mice showed significantly lowered percentages of normal sperms [(49.61±1.45)% and (53.22±3.51)%] as compared with the wild-type mice [(66.46±0.73)%; P<0.01] and significantly higher rates of sperm tail folding [(20.96±0.43)% and (20.73±0.21)%] than the wild-type mice [(5.74±0.72)%; P<0.01]. Ultrastructural observation of the sperm flagellum in Fscb/Cabyr--/-- and Fscb/Cabyr++/-- mice revealed that the longitudinal columns of the fibrous sheath disappeared in most of the sperms with disorganized circumferential ribs and heterogeneous reduction of the electron density in the sperms; some sperms showed disruption of the axoneme structure in the flagella with occasional disappearance and dissociation of the doublet microtubules at the center. ConclusionBoth Cabyr single gene knockout and Fscb/Cabyr double gene knockout can cause significant abnormalities in flagellum morphology and motility of the sperms in male mice. CABYR protein is essential for maintaining normal fibrous sheath structure and normal motility of the sperm flagella.
 

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更新日期/Last Update: 2020-12-22