Santiago-Moreno Julián, Toledano-Díaz Adolfo, Pulido-Pastor Antonio, Dorado Jesús, Gómez-Brunet Amelia, López-Sebastián Antonio
Departamento de Reproducción Animal, Avda. Puerta de Hierro Km 5,9, INIA, 28040 Madrid, Spain.
Theriogenology. 2006 Sep 15;66(5):1219-26. doi: 10.1016/j.theriogenology.2006.03.031. Epub 2006 May 2.
Tris-egg yolk based diluents provide adequate cryoprotection for the sperm of most wild species in which they have been tested. The objective of the current study was to evaluate various Tris-based diluents containing different concentrations of egg yolk, for the fertilizing ability of epididymal spermatozoa of the Spanish ibex (Capra pyrenaica) after freezing and thawing. For this purpose, we used heterologous in vivo fertilization by intrauterine insemination of domestic goats (Capra hircus). In Experiment 1, a Tris-citric acid-glucose (TCG) diluent containing 6% (v/v) egg yolk and a TCG extender containing 20% egg yolk were compared. In Experiment 2, a TCG-6% egg yolk extender was compared with Triladyl-20% egg yolk. Diluted samples were cooled slowly to 5 degrees C over 1 h and equilibrated at that temperature for 2 h. At that point, aliquots of samples were loaded into 0.25 ml straws, and frozen in nitrogen vapor for 10 min. The fertility of spermatozoa frozen in TCG-6% egg yolk was higher (P<0.05) than for those extended with TCG-20% egg yolk, and tended to be higher than for those frozen with Triladyl-20% egg yolk. From the results of this study, the use of Tris-based extenders containing low concentrations of egg yolk (6%) is recommended for cryopreserving Spanish ibex epididymal spermatozoa.
基于Tris-蛋黄的稀释剂能为大多数已测试的野生物种的精子提供足够的冷冻保护。本研究的目的是评估含有不同浓度蛋黄的各种基于Tris的稀释剂对西班牙羱羊(Capra pyrenaica)附睾精子冷冻和解冻后的受精能力。为此,我们通过对家山羊(Capra hircus)进行子宫内授精来进行异种体内受精。在实验1中,比较了含有6%(v/v)蛋黄的Tris-柠檬酸-葡萄糖(TCG)稀释剂和含有20%蛋黄的TCG稀释液。在实验2中,将TCG-6%蛋黄稀释液与Triladyl-20%蛋黄稀释液进行了比较。稀释后的样品在1小时内缓慢冷却至5℃,并在该温度下平衡2小时。此时,将等分的样品装入0.25毫升的细管中,并在氮蒸气中冷冻10分钟。用TCG-6%蛋黄冷冻的精子的受精能力高于用TCG-20%蛋黄稀释的精子(P<0.05),并且倾向于高于用Triladyl-20%蛋黄冷冻的精子。根据本研究的结果,建议使用含有低浓度蛋黄(6%)的基于Tris的稀释液来冷冻保存西班牙羱羊的附睾精子。