应用透射电镜技术研究了超低温冷冻保存对中间球海胆Strongylocentrotus intermedius精子超微结构的影响.结果表明:不加抗冻剂的情况下,精子线粒体和顶体结构不完整,质膜、鞭毛破损.加入保护剂后,多数精子结构完好,部分精子细胞器受损,表现为质膜褶皱或膨胀举起,与核膜的间隙明显增大；顶体肿胀、破裂或脱落；线粒体嵴间隙增大,内部出现空泡,甚至线粒体内膜破损,内嵴减少,呈弥散状；鞭毛被膜肿胀,呈波浪状,“9+2”型结构模糊；细胞核结构没有明显变化.应用单细胞凝胶电泳技术研究了精子冷冻保存前后DNA的损伤.结果表明,不加抗冻剂的冷冻组及添加抗冻剂的试验组精子DNA损伤情况与鲜精相比均无显著差异(P＞0.05),冷冻对精子结构的损伤是造成精子活力及受精率下降的主要原因.%Transmission electron microscopy was used to study the effects of cryopreservation on ultrastructure of sperm in sea urchin Strongylocentrotus intermedius. The results showed that the sperm was found to be disruption of mitochondria, acrosome and damage of plasma membrane, and flagellum of all the post-thawed sperm without addition of cryoprotectant. The perfect structure was observed in most of the post-thawed sperm exposed to the cryopro-tectant, only a few sperm being damaged, showing the following syndromes: the plasma membrane was swelled and broke off from the nuclear membrane; the acrosome swelled or lost; mitochondria cristae space enlarged, or the external membrane of mitochondrion broke, mitochondrial cristae declined; the membrane of flagellum swelled, the structure of "9+2" dimed in the unthawn damaged sperm; and the unchange nuclei. The single cell gel electropho-resis(SCGE) of DNA damage in the sea urchin sperm before and after cryopreservation revealed that there were no significant differences in the DNA damage of the fresh sperm between the control group in which the cryoprotectant was not added and the experimental group which the cryoprotectant was added (P>0. 05). The decline in sperm motility and fertilization rate was heavily attributed to the structural damage of the sperm in frozen states.
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