精子和卵母细胞质量控制对山羊卵胞质内精子注射(ICSI)受精的影响  

EFFECTS OF SPERM AND OOCYTE QUALITY CONTROL ON INTRACYTOPLASMIC SPERM INJECTION(ICSI)IN GOATS

作　　者：周佳勃[1] 吴延光[1] 韩东[1] 刘丽清[1] 谭秀文[1] 刘娜[1] 罗明玖[1] 常仲乐[1] 谭景和[1] 

机构地区：[1]山东农业大学动物科技学院

出　　处：《实验生物学报》2004年第5期367-374,共8页Acta Biologiae Experimentalis Sinica

基　　金：国家科技部"973"项目(编号G200016107);山东省良种产业化(三0)工程项目资助。

摘　　要：以体外成熟卵母细胞为材料研究了精子来源及制动处理方法、卵母细胞质量及注射后激活等因素对山羊ICSI效果的影响。结果说明,附睾头、体和尾精子ICSI后的受精率、卵裂率和桑椹胚/囊胚发育率与射出的鲜精精子都没有明显差异(p>0.05),但带下注射时附睾头和体精子的受精和发育率显著低于附睾尾和射精精子。在以4种不同方法致死的精子中,室温保存24h的死精子ICSI受精、卵裂和桑椹/囊胚率虽然低于对照组,但是明显高于其它方式致死的精子;5℃保存15天的死精子受精和发育效果最差。0.0005%Triton X-100处理精子的受精率、卵裂率和桑椹/囊胚率显著(p<0.05)高于制动对照组、不制动对照组和其它浓度组。经高渗处理法检测质量好的卵母细胞ICSI受精和胚胎发育效果显著好于质量差的卵母细胞。与对照组相比,A23187和Ionomycin/6-DMAP激活处理均显著(p<0.05)提高ICSI的受精率、卵裂率和桑椹/囊胚发育率。因此,精子在附睾内的成熟过程主要与其获得与卵质膜融合能力有关;精液保存方法对精子受精能力的损伤程度有很大差异;适当浓度的Triton X-100处理可模仿精子制动;卵母细胞质量是影响ICSI效果的重要因素;注射精子后激活卵母细胞能保证山羊ICSI的受精效果。Effects of sperm and oocyte quality control on the efficiency of ICSI of in vitro matured goat oocytes were studied in this paper.The results showed that when injected intracyto- plasmically,spermatozoa from caput,corpus and cauda epididymidis resulted in similar rates of fertilization,cleavage and morulae/blastocysts,but when injected subzonally,spermatozoa from ca- put and corpus gave rise to significantly lower rates of fertilization and embryo development than spermatozoa from the cauda epididymidis and ejaculates.When dead spermatozoa collected from semen that had been preserved in different ways were used for ICSI,those dead from liquid stor- age at 20℃ for 24 h gave rise to the best,but those dead from liquid storage at 5℃ for 15 days produced the poorest fertilization and embryo development.When spermatozoa were treated with different concentrations of Triton X-100 before ICSI,significantly higher rates of fertilization, cleavage and morulae/blastocysts were obtained with 0.0005% Triton X-100 than with other con- centrations and manual immobilization.Oocytes were classified as of good and poor qualities by treatment in hypertonic sucrose solution,and rates of fertilization and embryo development were significantly higher in the good than in the poor oocytes after ICSI.Post-injection activation of oocytes with either A23187 or ionomycin/6-DMAP significantly increased the rates of fertilization, cleavage and morulae/blastocysts after ICSI.It is therefore concluded that (i) epididymal matura- tion mainly endowed spermatozoa with the capacity to fuse with the egg plasma membrane;(ii) different methods of semen storage caused different impairment of sperm fertilizing capacity;(iii) pre-injection treatment of spermatozoa with proper concentrations of Triton X-100 might be used to replace manual immobilization for ICSI;(iv)oocyte quality was a major factor influencing the efficiency of ICSI;(v)post-injection activation treatment of oocytes improved fertilization and em- bryo development after ICSI.

关 键 词：ICSI 精子 对照组 注射 受精 效果 射精 胞质 卵母细胞 山羊 

分 类 号：Q813[生物学—生物工程]