Thuwanut Paweena, Thongphakdee Ampika, Sommanustweechai Angkana, Siriaroonrat Boripat, Chatdarong Kaywalee
Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand.
J Vet Med Sci. 2013 Jan 31;75(1):123-5. doi: 10.1292/jvms.11-0491. Epub 2012 Oct 11.
In the present study, the quality of frozen-thawed epididymal and testicular sperm recovered from a Siamese Eld's deer was examined. The epididymal sperm quality was assessed in fresh, cold-stored at 4°C and frozen-thawed samples. Zona binding ability of the frozen-thawed epididymal samples with Burmese Eld's deer oocytes was also evaluated. Testicular sperm extracted from tissues frozen at -80 or -196°C for one month were examined for membrane and DNA integrity. Epididymal sperm retained their quality for up to 24 hr of cold storage at 4°C. The percentages of sperm motility, intact membrane, intact acrosome and intact DNA were 30, 46.5, 27 and 89.5% in the frozen and thawed epididymal sperm, and the average ability to bind with oocytes was 92.5 ± 64 sperm/oocytes. Around 70% of the sperm extracted from testicular tissues cryopreserved at -196 and -80°C for one month showed an intact membrane. In conclusion, epididymal and testicular sperm survived for more than 13 hr post-mortem. Furthermore, cold storage at 4°C and cryopreservation at -196 and -80°C maintain the quality of epididymal and testicular sperm. This study represents a model for male gamete rescue in endangered Eld's deer.
在本研究中,对从暹罗坡鹿回收的冻融附睾精子和睾丸精子的质量进行了检测。对新鲜、4℃冷藏和冻融样本中的附睾精子质量进行了评估。还评估了冻融附睾样本与缅甸坡鹿卵母细胞的透明带结合能力。对从-80℃或-196℃冷冻保存一个月的组织中提取的睾丸精子的膜完整性和DNA完整性进行了检测。附睾精子在4℃冷藏长达24小时仍能保持其质量。冻融附睾精子的精子活力、完整膜、完整顶体和完整DNA的百分比分别为30%、46.5%、27%和89.5%,与卵母细胞结合的平均能力为92.5±64个精子/卵母细胞。从-196℃和-80℃冷冻保存一个月的睾丸组织中提取的精子,约70%显示膜完整。总之,附睾精子和睾丸精子在死后存活超过13小时。此外,4℃冷藏以及-196℃和-80℃冷冻保存可维持附睾精子和睾丸精子的质量。本研究为濒危坡鹿的雄配子拯救提供了一个模型。