在提出的研究中,ROCK抑制剂Y-27632,抗冻蛋白III,研究了两种不同剂量的硼对冻融后安卡拉巴克精液的精子学参数的影响。在繁殖季节使用电子射精器从雄鹿中收集射精。将表现出适当特征的射精物合并并用于精液的稀释和冷冻。通过添加两种不同剂量的三种不同添加剂(ROCK抑制剂Y-27632,5和20µM;抗冻蛋白III,1和4µg/mL;硼,0.25和1mM)至对照扩展器。在35-37°C下用不同的增量剂稀释精液并装入吸管中。在液氮蒸气中冷冻的精子样本,在平衡之后,储存在液氮中。观察到,补充剂可改善冻融后安卡拉雄鹿精液的解冻后运动。对于5和10µM剂量的ROCK抑制剂(71.82%和74.04%的运动性),差异显着(p<0.01),以及0.25和1mM剂量的硼(76.36%和72.08%的运动性),与对照组相比(66.15%运动性)。关于冻融后顶体完整性和线粒体活性的评估,虽然补充剂在所有剂量下都能提供保护,疗效无统计学意义(p>0.05)。观察到,与对照组相比,在1µg/mL(1.23%±0.23%)的抗冻蛋白III和在所有剂量(0.25mM:1.83%和1mM:1.18%)的硼改善了DNA损伤(3.37%)(p<0.01),在解冻过程之后。在本研究中,已确定,添加到补充剂中的一些添加剂在解冻后可显著改善降压精子的运动性和DNA损伤。
In the presented study, the effects of ROCK inhibitor Y-27632, antifreeze protein III, and boron at two different doses were investigated on the spermatological parameters of Ankara buck semen after freeze−thawing. Ejaculates were collected from bucks using an electroejaculator during the breeding season. The ejaculates that showed appropriate characteristics were pooled and used in the dilution and freezing of semen. The extender groups were formed by adding two different doses of three different additives (ROCK inhibitor Y-27632, 5 and 20 µM; antifreeze protein III, 1 and 4 µg/mL; boron, 0.25 and 1 mM) to the control extender. The semen was diluted with the different extenders at 35−37 °C and loaded into straws. Sperm samples frozen in liquid nitrogen vapors, following equilibration, were stored in liquid nitrogen. It was observed that extender supplementation improved post-thaw motility of Ankara buck semen after freeze−thawing. Differences were significant (p < 0.01) for 5 and 10 µM doses of ROCK inhibitor (71.82% and 74.04 % motility), as well as for 0.25 and 1 mM doses of boron (76.36% and 72.08% motility), compared to the control group (66.15% motility). With respect to the evaluation of acrosomal integrity and mitochondrial activity after freeze−thawing, although supplementation provided protection at all doses, the efficacy was not statistically significant (p > 0.05). It was observed that DNA damage was improved by antifreeze protein III at 1 µg/mL (1.23% ± 0.23%) and by boron at all doses (0.25 mM: 1.83% and 1 mM: 1.18%) compared to the control group (3.37%) (p < 0.01), following the thawing process. In the present study, it was determined that some additives added to the extender provided significant improvements in buck spermatozoa motility and DNA damage after thawing.