川陕哲罗鲑卵子和精子的显微与超微结构观察

Microstructure and ultrastructure observation of eggs and sperms of Hucho bleekeri

  • 摘要:目的】观察川陕哲罗鲑正常卵子和精子的形态学结构,并比较不同活力精子的超微结构差异,揭示川陕哲罗鲑配子超微结构损伤与其活力下降的内在关联,为优化其人工繁育技术提供理论依据。【方法】以人工养殖性成熟的雌、雄性川陕哲罗鲑为研究对象,通过组织学结合扫描电子显微镜及透射电子显微镜对川陕哲罗鲑正常卵子和精子的形态结构进行观察,同时结合精子活力检测系统比对不同活力精子的超微结构差异。【结果】川陕哲罗鲑正常卵子的核膜溶解,核仁完全消失,在靠近卵膜位置分布有液泡;卵膜表面平整,壁孔大小均匀,受精孔前庭区平坦,精孔管较深,属于II型。川陕哲罗鲑正常精子的结构属于鞭毛型,由头部、中段和尾部3个部分组成;头部呈椭圆形或近圆形,无顶体结构,细胞核内可见空隙(核泡),其染色质呈颗粒状或均质状;中段(颈部)较短,其主要结构包括中心粒复合体及袖套;尾部(鞭毛)细长光滑,鞭毛轴丝具有典型的“9+2”微管结构。扫描电子显微镜观察发现,无活力川陕哲罗鲑精子的头部轮廓不规则、皱缩塌陷,中段连接结构不明显,鞭毛普遍出现断裂和缺失,无法维持正常形态;高活力精子的形态结构完整,头部呈饱满的卵圆形,中段连接紧密,鞭毛细长且无明显损伤,各部分结构清晰。透射电子显微镜观察结果显示,无活力川陕哲罗鲑精子的线粒体结构被破坏,膜膨胀变大破损,线粒体内部电子密度下降,头部表面褶皱不平滑,核膜破裂或消失,染色质松散甚至解体;高活力精子的细胞质致密、均匀饱满,细胞质膜完整,结构正常。【结论】川陕哲罗鲑正常卵子的卵膜表面平整,其受精孔属于II型,核仁完全消失,在靠近卵膜位置分布有液泡;正常精子由头部、中段及鞭毛3个部分构成,具备硬骨鱼类典型的无顶体结构特征,鞭毛轴丝为“9+2”微管排列模式。质膜、核膜、线粒体、鞭毛结构完整性及成熟度直接关联川陕哲罗鲑的精子活力,是评价川陕哲罗鲑精液质量及优化其人工繁育技术的关键形态学指标。

     

    Abstract:Objective】This study aimed to observe the normal morphological structure of eggs and sperms of Hucho bleekeri, compare differences in ultrastructure of sperms of varying motility, and reveal the intrinsic association between gamete ultrastructure damage and decreased motility in Hucho bleekeri, providing theoretical basis for optimizing its artificial breeding techniques.【Method】Sexually mature female and male Hucho bleekeri reared under artificial culture conditions were used as research subjects. The morphological structure of normal eggs and sperms in Hucho bleekeri was observed through histological analysis with scanning electron microscope and transmission electron microscope. Meanwhile, the differences in ultrastructure of sperms of varying motility were compared using the sperm motility detection system.【Result】In normal eggs of Hucho bleekeri, the nuclear membrane was dissolved, the nucleoli completely disappeared, and vacuoles distributed close to egg membrane. The egg membrane surface was smooth, the pores were uniform in size, the vestibular region of micropyle was flat, and the micropylar canal was relatively deep, belonging to type II. The structure of normal sperms in Hucho bleekeri was of flagellate type, consisting of three parts (head, midpiece, and tail). The head was elliptical or nearly rounded, without acrosomal structure, and intranuclear space (nuclear vacuole) was visible within the nucleus, with granular or homogeneous chromatins. The midpiece (neck) was short, consisting mainly of the centriolar complex and sleeve. The tail (flagellum) was slender and smooth, with the flagellar axoneme exhibiting the typical “9+2” microtubule arrangement. Scanning electron microscope observations revealed that immotile Hucho bleekeri sperms displayed irregular head contours that were shrunken and collapsed, with an inconspicuous midpiece connecting structure, and the flagella were commonly broken and missing, unable to maintain normal morphology; highly motile sperms exhibited intact structure morphology, with plump ovoid heads, a tightly connected midpiece, slender flagella without obvious damage, and clear structures in all parts. Transmission electron microscope observations showed that in immotile Hucho bleekeri sperms, the mitochondrial structure was disrupted, the membranes were swollen, enlarged, and ruptured, and the internal electron density of mitochondria was decreased, with a wrinkled and uneven head surface, disrupted or disappeared nuclear membrane, and loose and even disintegrated chromatins; in highly motile sperms, the cytoplasm was dense, uniform, and compact, and the plasma membrane was intact with normal structure.【Conclusion】The oolemma of normal eggs in Hucho bleekeri is smooth, and the micropyle belongs to type II, with nucleoli completely disappeared and vacuoles distributed adjacent to egg membrane; normal sperms consist of three parts: the head, midpiece, and flagellum, exhibiting the typical acrosome-free structural characteristic of teleost fish, with the flagellar axoneme in a “9+2” microtubule arrangement. The structure integrity and maturity of the plasma membranes, nuclear membranes, mitochondria, and flagellar structure are directly associated with sperm motility in Hucho bleekeri, serving as key morphological indicators for evaluating semen quality and optimizing artificial breeding techniques.

     

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