Silva Andréia M, Praxedes Erica C G, Campos Lívia B, Bezerra Luana G P, Moreira Samara S J, Maia Keilla M, Souza Ana L P, Silva Alexandre R
Laboratory of Animal Germplasm Conservation (LCGA), Universidade Federal Rural do Semi-Árido (UFERSA), Mossoró, RN, Brazil.
Laboratory of Animal Germplasm Conservation (LCGA), Universidade Federal Rural do Semi-Árido (UFERSA), Mossoró, RN, Brazil.
Anim Reprod Sci. 2018 Apr;191:64-69. doi: 10.1016/j.anireprosci.2018.02.009. Epub 2018 Feb 15.
As a non-threatened hystricognath rodent species, Spix's yellow-toothed cavies can be used as a model for the development of assisted reproductive techniques for the conservation of closely related species. The objective was to establish a functional protocol for cryopreservation of epididymal sperm from these cavies. Twelve sexually mature males, ∼2 y old and weighing ∼300 g, were euthanized. Sperm were recovered by retrograde flushing of the vas deferens and cauda epididymis with Tris extender. Thereafter, sperm were extended in Tris plus 20% egg yolk, with 3%, 6% or 9% glycerol or dimethyl sulfoxide (DMSO), placed in 0.25 mL straws and cryopreserved in liquid nitrogen. Sperm concentration, motility (using computer-assisted sperm analysis; CASA), plasma membrane integrity, osmotic response, morphology and sperm binding-ability were determined in fresh and frozen-thawed sperm. For most sperm endpoints, glycerol was a more desirable cryoprotectant than DMSO. Data (mean ± SEM) were similar with use of 3%, 6%, and 9% glycerol (P > 0.05) in osmotic response (40.66 ± 6.3%, 42.5 ± 7.1%, and 39.5 ± 5.0% respectably), and membrane integrity (55.17 ± 5.5%, 68.4 ± 4.1%, and 59.1 ± 4.9% respectably). Among concentrations assessed, the use of 6% glycerol resulted in the greatest (P < 0.05) post-thaw values for total motility (60.9 ± 4.4%), rapid subpopulation motility (27.7 ± 3.1%) and sperm-binding capability (227.0 ± 20.2). In conclusion, epididymal sperm from the Spix's yellow-toothed cavies (G. spixii) are optimally cryopreserved in Tris extender with 6% glycerol and 20% egg yolk.
作为一种未受威胁的豪猪亚目啮齿动物物种，斯皮克斯黄齿豚鼠可作为开发辅助生殖技术以保护近缘物种的模型。目的是建立一套用于冷冻保存这些豚鼠附睾精子的有效方案。对12只性成熟雄性豚鼠实施安乐死，这些豚鼠约2岁，体重约300克。通过用Tris稀释液逆行冲洗输精管和附睾尾部来采集精子。此后，将精子用含20%蛋黄的Tris稀释，并添加3%、6%或9%的甘油或二甲基亚砜（DMSO），放入0.25毫升细管中并在液氮中冷冻保存。对新鲜和冻融后的精子测定精子浓度、活力（使用计算机辅助精子分析；CASA）、质膜完整性、渗透反应、形态和精子结合能力。对于大多数精子指标，甘油是比DMSO更理想的冷冻保护剂。在渗透反应（分别为40.66±6.3%、42.5±7.1%和39.5±5.0%）和膜完整性（分别为55.17±5.5%、68.4±4.1%和59.1±4.9%）方面，使用3%、6%和9%甘油的数据（平均值±标准误）相似（P>0.05）。在所评估的浓度中，使用6%甘油解冻后总活力（(60.9±4.4%）、快速亚群活力（27.7±3.1%）和精子结合能力（227.0±20.2）的值最高（P<0.05）。总之，斯皮克斯黄齿豚鼠（G. spixii）的附睾精子在含6%甘油和20%蛋黄的Tris稀释液中冷冻保存效果最佳。
