波尔山羊精子体外获能及获能前后超微结构的变化

【作者】 斛智莲；

【导师】 岳文斌；

【作者基本信息】 山西农业大学 ， 动物遗传育种与繁殖， 2001， 硕士

【摘要】 本文就国内新引进的波尔山羊精子体外获能及获能前后超微结构的变化进行研究。获能处理前先进行活精子分离，然后按实验之需，用含有肝素的培养液获能处理，获能结束后用溶血磷脂酰胆碱（LC）诱导顶体反应，TG染色法测得有顶体反应的活精子作为评价精子获能的指标。肝素添加剂量设5个水平，即0μg/ml、5μg/ml、10μg/ml、15μg/ml、20μg/ml；处理时间分4h、3h、2h、1h、15min5个水平；LC处理时间设0min、3min、6min、9min、12min、15min、20min7个水平。电镜样品用常规方法固定、脱水、包埋、聚合，LKB-2088V切片机切片，铀铅片染，JEM-100XII观察，拍照。 结果表明：三种培养液中，从获能效果、精子活力和活精子百分率来考虑，S培养液效果最好，4h的顶体反应率达60.88％，与D、B培养液的获能效果相比，差异极显著（P＜0.01），获能4h和6h的效果差异不显著（P＞0.05）；肝素的添加剂量以15μg/ml和20μg/ml的效果最好，与其它处理组的效果相比差异显著（P＜0.05），但这两个水平间的效果差异不显著（P＞0.05）；LC诱导顶体反应的时间在15min时效果极显著高于15min以下处理组的效果（P＜0.01）。羊精子的超微结构与其它哺乳动物的类似，分头、颈、尾三部分。头部主要由细胞核构成，含有遗传物质；颈部脆弱，尾部易从此脱落；尾部由中段、主段和末段组成。精子获能后，顶体质膜膨胀、断裂和丢失；顶体外膜囊泡化，进而顶体内膜裸露。更多还原

【Abstract】 IN VITRO CAPACITAT ION OF BOER SPERM AND ITS ULTRU?STRACTURE BEFORE OR AFTER CAPACITAT ION Abstract : The objectives of this study were to select the excellent culture media lii vitro capacitafion and observe the fine structure of Boer sperm before or after capacitatiom Three cultwe media used are 5, D and B. Separate the motile spennatozoa before capacitation, and lhen exposure capacitated sperm to lysophosphatidyl-choline(LC) to induce acmsome reaction. Capacitation was evaluated by determining the number of motile aciosome-reacted sperm using TG staining. Sperm were incubated 4h by S media of0 g/ml 5p.g/mU. 10 jig/mb 15 jig/mi-. 2Ojig/mlheparin,andheparinwasaddedat 0h 1W. 2W. 3W. 3.75hafterthe start of incubation. Atthe end of incubation, Oor3Op.gIml LC was added, and the percentage of sperm acrosome-reacted was determined by TG staining after a thither 1 5mm incubation. Obtain the sperm capacitated or non-capacitated and process according to routine procedures for TEM. Results indicated S media was the best culture media allowing for its effects on capacitation, motility and viability of sperm. The percentage of acrosome-reacted sperm at 4h was significantly different from other t media (P < 0.01) . Supplements of 1 5jig-l or 20 jig/mI heparin had apparent effect on acrosome reaction of sperm (P < 0.05) . But there was no apparent difference between these two treatments (P >0.05). Induce capacitated sperm with LC for a further 15mm and obtain the results of acmsome-reacted sperm which was significantly different from any other treatments (P < 0.01) . But there were difference among individuals. The ultra-stmcture of Boer spermatozoa was similar to that of other mammals. Eveiy sections of sperm had its ox characteristics. The great morphological changes had happened after capacitation. and that maybe have significant importance in fertilization.更多还原