López-Saucedo J, Paramio M T, Fierro R, Izquierdo D, Catalá M G, Coloma M A, Toledano-Díaz A, López-Sebastián A, Santiago-Moreno J
Fertility Center Cancún, Cancún, Mexico.
Departamento de Ciencia Animal y de los Alimentos, Facultad de Veterinaria, Universidad Autónoma de Barcelona, 08193 Bellaterra, Barcelona, Spain.
Cryobiology. 2014 Jun;68(3):389-94. doi: 10.1016/j.cryobiol.2014.03.009. Epub 2014 Apr 1.
The aim of this work was to evaluate the protective effect of catalase (CAT) on frozen/thawed ibex epididymal sperm recovered post mortem, and to detect any harmful effect this might have on sperm fertilisation capacity. Epididymal spermatozoa were diluted using a Tris-citric acid-glucose medium (TCG) composed of 3.8% Tris (w/v), 2.2% citric acid (w/v), 0.6% glucose (w/v), 5% glycerol (v/v), and 6% egg yolk (v/v). Sperm masses from the right epididymis were diluted with TCG medium, while those from the left were diluted with TCG medium supplemented with 200IU/mL CAT. Heterologous in vitro fertilisation (IVF) was used to assess the fertilisation capacity of this sperm. The addition of CAT to the extender did not improve frozen/thawed sperm variables. Moreover, a reduced fertilisation capacity was detected: sperm diluted with TCG provided 25.5% 2PN zygotes, while just 13.2% was recorded for that diluted with TCG-CAT (P<0.01). The percentage of cleaved embryos at 48hpi was higher (P<0.01) with the TCG sperm than with the TCG-CAT sperm (16.7% vs. 7.6%). The use of 200IU/mL CAT as an additive cannot, therefore, be recommended for the preservation of ibex epididymal sperm. Other antioxidants should, however, be tested in both this and related wild mountain ungulates.
本研究旨在评估过氧化氢酶(CAT)对死后回收的冻融野生山羊附睾精子的保护作用,并检测其对精子受精能力可能产生的任何有害影响。附睾精子用由3.8%三羟甲基氨基甲烷(w/v)、2.2%柠檬酸(w/v)、0.6%葡萄糖(w/v)、5%甘油(v/v)和6%蛋黄(v/v)组成的三羟甲基氨基甲烷 - 柠檬酸 - 葡萄糖培养基(TCG)进行稀释。右侧附睾的精子团用TCG培养基稀释,而左侧的精子团用添加了200IU/mL CAT的TCG培养基稀释。采用异源体外受精(IVF)来评估这种精子的受精能力。向稀释液中添加CAT并没有改善冻融精子的各项指标。此外,还检测到受精能力下降:用TCG稀释的精子产生了25.5%的双原核受精卵,而用TCG - CAT稀释的精子仅为13.2%(P<0.01)。在48小时受精卵期,TCG精子的胚胎裂解率高于TCG - CAT精子(16.7%对7.6%,P<0.01)。因此,不建议使用200IU/mL CAT作为添加剂来保存野生山羊附睾精子。然而,应该在这种以及相关的野生山地有蹄类动物中测试其他抗氧化剂。