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[精子和卵母细胞质量控制对山羊卵胞浆内单精子注射(ICSI)的影响]

[Effects of sperm and oocyte quality control on intracytoplasmic sperm injection (ICSI) in goats].

作者信息

Zhou Jia Bo, Wu Yan Guang, Han Dong, Liu Li Qing, Tan Xiu Wen, Liu Na, Luo Ming Jiu, Chang Zhong Le, Tan Jing He

机构信息

(College of Anim. Sci. and Technology, Shandong Agricultural University, Taian.

出版信息

Shi Yan Sheng Wu Xue Bao. 2004 Oct;37(5):367-74.

Abstract

ABSTRACT 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 intracytoplasmically, spermatozoa from caput, corpus and cauda epididymidis resulted in similar rates of fertilization, cleavage and morulae/blastocysts, but when injected subzonally, spermatozoa from caput 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 storage at 20 degrees C for 24 h gave rise to the best, but those dead from liquid storage at 5 degrees C 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 concentrations 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 maturation 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 embryo development after ICSI.

摘要

摘要 本文研究了精子和卵母细胞质量控制对体外成熟山羊卵母细胞胞浆内单精子注射(ICSI)效率的影响。结果表明,将附睾头、体、尾的精子注入卵母细胞胞浆时,受精、卵裂及桑葚胚/囊胚形成率相似,但将精子注入卵周隙时,附睾头和体的精子受精率和胚胎发育率显著低于附睾尾精子和射出精子。用不同保存方式收集的死精子进行ICSI时,20℃液体保存24小时的死精子效果最佳,而5℃液体保存15天的死精子受精和胚胎发育最差。ICSI前用不同浓度的Triton X-100处理精子,0.0005% Triton X-100组的受精、卵裂及桑葚胚/囊胚形成率显著高于其他浓度组及手工固定组。用高渗蔗糖溶液处理将卵母细胞分为优质和劣质,ICSI后优质卵母细胞的受精和胚胎发育率显著高于劣质卵母细胞。用A23187或离子霉素/6-二甲氨基嘌呤对卵母细胞进行注射后激活,可显著提高ICSI后的受精、卵裂及桑葚胚/囊胚形成率。因此得出结论:(i)附睾成熟主要赋予精子与卵质膜融合的能力;(ii)不同的精液保存方法对精子受精能力的损害不同;(iii)ICSI前用适当浓度的Triton X-100处理精子可替代手工固定;(iv)卵母细胞质量是影响ICSI效率的主要因素;(v)卵母细胞注射后激活处理可提高ICSI后的受精和胚胎发育。

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