4℃液态保存对奶山羊精子超微结构的影响

Effect of 4℃Liquid Storage on Ultrastructure of Dairy Goat Spermatozoa

旨在探究4℃液态保存中奶山羊精子超微结构变化,为阐明影响奶山羊精子液态保存质量的相关机制提供形态学依据。采集关中奶山羊精液进行4℃液态保存,伊红染色检测保存至0、24、48、72、96、120和144 h的精子活率,扫描电镜和透射电镜观察0、48、96 h的精子超微结构。伊红染色检测结果发现,精子活率随液态保存时间延长逐渐降低。超微结构显示,0 h时,精子形态结构正常、顶体完整光滑、精子尾部线粒体鞘结构完整,线粒体排列规则、形态大小正常;48 h时,部分精子出现尾部断裂卷曲、核膜膨胀折叠和线粒体肿胀等结构损伤;96 h时,部分精子出现顶体破损、主段断裂、核崩解碎裂、线粒体缺失和排列混乱等损伤。48与96 h均有部分精子出现典型凋亡形态特征,如:核表面针状突起、囊性空泡和凋亡小体。以上结果表明,4℃液态保存过程中,奶山羊精子出现结构损伤,部分精子具有典型凋亡形态特征,精子活率逐渐下降,提示奶山羊精子结构损伤导致活率下降。

To explore the ultrastructure changes of dairy goat spermatozoa during liquid storage at 4℃,and provide a morphological basis for elucidating the relevant mechanisms that affect the quality of dairy goat spermatozoa during liquid storage.The semen of Guanzhong dairy goats was collected and stored in a liquid state at 4℃,sperm viability at 0,24,48,72,96,120 and 144 h of storage was detected by eosin staining,the ultrastructure of spermatozoa at 0,48,and 96 h was observed by scanning electron microscope and transmission electron microscope.The results of eosin staining showed that sperm viability gradually decreased with the prolongation of liquid storage time.The ultrastructure showed that at 0 h,the sperm morphology and structure were normal,the acrosome was intact and smooth,the mitochondrial sheath of sperm tail were intact,and the mitochondria were arranged regularly,and the morphology and size were normal;at 48 h,some spermatozoa showed structural damage such as tail breakage and curling,nuclear membrane expansion and folding,and mitochondrial swelling;at 96 h,some spermatozoa suffered damages such as acrosome damage,main segment fracture,nuclear disintegration and fragmentation,mitochondrial deletion and disordered arrangement.At 48 and 96 h,some spermatozoa exhibited typical apoptotic morphological features,such as needle-like protrusions on the nuclear surface,cystic vacuoles and apoptotic bodies.The above results showed that during the liquid storage at 4℃,the sperm of dairy goats suffered structural damage,some spermatozoa had typical apoptotic morphological characteristics,sperm viability decreased gradually,suggesting that the sperm structure damage of dairy goats led to the decrease of viability.