Veterinary Clinical Department, University of Bologna, Ozzano Emilia (BO), Italy.
Theriogenology. 2010 Apr 15;73(7):886-92. doi: 10.1016/j.theriogenology.2009.11.012. Epub 2010 Jan 13.
An optimal protocol for cat semen cryopreservation has not yet been defined. Addition of Equex STM Paste has been tested for epididymal cat spermatozoa but not for ejaculated cat spermatozoa. Furthermore, the effect of Equex STM Paste on fertilizing ability of cryopreserved semen has never been evaluated in that species. Therefore, the aims of the current study were to investigate if addition of Equex STM Paste to a freezing extender for electroejaculated cat (Felis catus) semen would improve postthaw sperm quality and if sperm fertilizing ability after cryopreservation with or without Equex STM Paste was preserved. Semen was collected by electroejaculation and frozen in a Tris-glucose-citrate egg yolk extender supplemented with (0.5% vol/vol) or without Equex STM Paste. In Experiment 1, sperm motility, membrane integrity, and acrosomal status were determined immediately after collection and at 0, 3, and 6h postthaw. In Experiment 2, frozen semen from the two groups was used for in vitro fertilization (IVF) of in vitro-matured cat oocytes. Cleavage rate was recorded 30h after IVF, and embryo development was evaluated on Days 6 and 7 of culture. In Experiment 1, the rate of motile spermatozoa after freezing-thawing was higher when Equex STM Paste was added to the freezing extender, but progressive motility score was not influenced (P>0.05). Sperm membrane integrity was positively affected (P<0.05) by the addition of the detergent. Intact acrosomes after thawing were similar (P>0.05) between groups. Even if the decreasing rates of motility and membrane integrity were more rapid in presence of Equex than those in controls, total motility and sperm viability were similar at 3 and 6h after thawing (P>0.05). In Experiment 2, there was no difference in fertilizing ability and embryo development between the two groups (P>0.05). The results of this study demonstrate that the addition of Equex STM Paste in the freezing extender avoids the loss of motile spermatozoa and maintains fertilizing ability of frozen-thawed spermatozoa.
尚未确定猫精液冷冻保存的最佳方案。已经针对附睾猫精子进行了添加 Equex STM 膏的试验,但尚未针对射出的猫精子进行试验。此外,Equex STM 膏对冷冻精液受精能力的影响在该物种中从未得到评估。因此,本研究的目的是研究在电刺激猫(Felis catus)精液的冷冻液中添加 Equex STM 膏是否会提高解冻后精子的质量,以及在添加或不添加 Equex STM 膏的情况下冷冻保存后精子的受精能力是否得到保持。通过电刺激法采集精液,并在含有(0.5%体积/体积)或不含有 Equex STM 膏的 Tris-葡萄糖-柠檬酸卵黄液冷冻液中冷冻。在实验 1 中,在采集后立即以及解冻后 0、3 和 6 小时测定精子的运动性、膜完整性和顶体状态。在实验 2 中,来自两组的冷冻精液用于体外成熟的猫卵母细胞的体外受精(IVF)。在 IVF 后 30 小时记录卵裂率,并在培养的第 6 和第 7 天评估胚胎发育情况。在实验 1 中,添加 Equex STM 膏可提高冷冻-解冻后精子的运动率,但对精子的前向运动评分没有影响(P>0.05)。精子膜完整性受到洗涤剂的积极影响(P<0.05)。解冻后的完整顶体在各组之间相似(P>0.05)。即使在含有 Equex 的情况下,精子的运动和膜完整性的下降速度比对照组更快,但在解冻后 3 和 6 小时,总运动率和精子活力相似(P>0.05)。在实验 2 中,两组的受精能力和胚胎发育没有差异(P>0.05)。本研究的结果表明,在冷冻液中添加 Equex STM 膏可避免运动精子的损失,并保持冷冻-解冻精子的受精能力。