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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Morphological phenotypes and changes in motility parameters of sperms in
Fscb/Cabyr double knockout mice- 第三军医大学学报 2020年42卷第24期 页码：2358-2365
- 作者机构：
1.400042 重庆，陆军军医大学(第三军医大学)大坪医院泌尿外科
2..400038 重庆，陆军军医大学(第三军医大学)预防医学院毒理学研究所
3.463000 河南 驻马店，中国人民解放军第990医院泌尿外科
- 作者简介：
[ 李彦锋，E-mail:1738401063@qq.com LI Yanfeng, E-mail:1738401063@qq.com
- 基金信息：国家自然科学基金面上项目(81571491)
DOI：10.16016/j.1000-5404.202007083
中图分类号： R322.1;R329.24;R394-33收稿：2020-05-06，
修回：2020-06-05，
纸质出版：2020-12-30
- 稿件说明：
移动端阅览
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.Zhongtai LI, Yanfeng LI, Jinyi LIU, et al. Morphological phenotypes and changes in motility parameters of sperms in
Fscb/Cabyr double knockout mice[J]. Journal of Third Military Medical University, 2020, 42(24): 2358-2365.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. DOI： 10.16016/j.1000-5404.202007083.Zhongtai LI, Yanfeng LI, Jinyi LIU, et al. Morphological phenotypes and changes in motility parameters of sperms in
Fscb/Cabyr double knockout mice[J]. Journal of Third Military Medical University, 2020, 42(24): 2358-2365. DOI： 10.16016/j.1000-5404.202007083.
摘要
目的
2
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
2
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
2
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显著低于野生型小鼠；其精子尾折叠比例显著高于野生型小鼠。
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
2
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
Objective
2
To investigate the changes in morphological characteristics of sperm flagellum and in sperm motility parameters in
Fscb/Cabyr
double gene knockout (
Fscb/Cabyr
--/--
) mice.
Methods
2
The 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 acroso
me 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.
Results
2
No 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 1.45)% and (53.22±3.51)%
]
as compared with the wild-type mice and significantly higher rates of sperm tail folding
]
than the wild-type mice. 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.
Conclusion
2
Both 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.
关键词
Keywords
references
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