Chatdarong K, Thuwanut P, Manee-in S, Lohachit C, Axnér E
Department of Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Pathumwan, Bangkok, Thailand.
Reprod Domest Anim. 2010 Apr;45(2):221-7. doi: 10.1111/j.1439-0531.2008.01218.x. Epub 2008 Dec 15.
The present study aimed to compare cat sperm quality after thawing using two different temperatures (37 and 70 degrees C) and to investigate the effects of post-thaw dilution on the sperm quality and longevity of ejaculated cat spermatozoa. Six ejaculates of each of six male cats were collected using an electroejaculator (total 36 ejaculates). The semen was frozen in 0.25-ml straws using a Tris egg yolk extender containing Equex STM paste. Four straws prepared from each ejaculate were thawed at four different occasions; (i) at 37 degrees C for 15 s, (ii) at 37 degrees C for 15 s and diluted 1 : 2 with Tris buffer (v/v), (iii) at 70 degrees C for 6 s, (iv) at 70 degrees C for 6 s and diluted 1 : 2 with Tris buffer (v/v). The percentages of motile spermatozoa, the scores of progressive motility, the percentages of spermatozoa with intact plasma membrane (using SYBR-14/EthD-1 stains) and intact acrosome (using fluorescein isothiocyanate conjugated peanut agglutinin/propidium iodide stains) were evaluated in fresh semen at 0, 2, 4 and 6 h after thawing. The thawing temperature had no effect on any sperm parameters throughout the incubation period (p > 0.05). The dilution after thawing improved sperm motility, progressive motility and acrosome integrity (p < 0.05). The thawing of cat spermatozoa and subsequently diluting with Tris buffer resulted in an immediate (at 0 h) overall (combined over temperature) percentage of motile sperm of 64.8 +/- 10.7 (mean +/- SD), a score of progressive motility of 4.0 +/- 0.5, a percentage of spermatozoa with intact plasma membrane of 64.4 +/- 12.1 and intact acrosome of 44.8 +/- 20.2. In conclusion, frozen cat semen can be thawed either at 37 or 70 degrees C and post-thaw dilution is recommended to reduce the toxic effect of some ingredients in the extender during post-thaw incubation.
本研究旨在比较使用两种不同温度(37℃和70℃)解冻后的猫精子质量,并研究解冻后稀释对射精的猫精子质量和寿命的影响。使用电刺激采精器收集6只雄猫的精液,每只猫采集6次射精(共36次射精)。精液使用含有Equex STM糊剂的Tris蛋黄稀释液冷冻于0.25毫升细管中。每次射精制备的4根细管在4个不同时间点解冻:(i)在37℃解冻15秒;(ii)在37℃解冻15秒,并用Tris缓冲液按1:2稀释(v/v);(iii)在70℃解冻6秒;(iv)在70℃解冻6秒,并用Tris缓冲液按1:2稀释(v/v)。在解冻后0、2、4和6小时对新鲜精液中活动精子的百分比、前进运动评分、具有完整质膜的精子百分比(使用SYBR-14/碘化丙啶染色)和具有完整顶体的精子百分比(使用异硫氰酸荧光素偶联花生凝集素/碘化丙啶染色)进行评估。在整个孵育期,解冻温度对任何精子参数均无影响(p>0.05)。解冻后稀释可改善精子活力、前进运动和顶体完整性(p<0.05)。猫精子解冻并随后用Tris缓冲液稀释后,活动精子的即时(0小时)总体(综合不同温度)百分比为64.8±10.7(平均值±标准差),前进运动评分为4.0±0.5,具有完整质膜的精子百分比为64.4±12.1,具有完整顶体的精子百分比为44.8±20.2。总之,冷冻的猫精液可以在37℃或70℃解冻,建议解冻后稀释以减少解冻后孵育期间稀释液中某些成分的毒性作用。