Younis A I, Rooks B, Khan S, Gould K G
Division of Molecular Medicine, Yerkes Regional Primate Research Center, Emory University, Atlanta, Georgia 30329, USA.
J Androl. 1998 Mar-Apr;19(2):207-14.
We investigated the effects of antifreeze peptides (AFP) and insulin transferrin selenium (ITS) on the motility and membrane integrity of chimpanzee (Pan troglodytes) spermatozoa after chilling (0-5 degrees C) and thawing. The effects of three thawing procedures, in the presence or absence of AFP and ITS, on sperm motility and on the status of the plasma membrane and acrosome were also examined. During chilling, AFP and ITS seem mildly cytotoxic, as the progressive motility and velocity (curvilinear and straight line) declined significantly at AFP concentrations of 1, 10, and 100 microg/ml and at ITS concentrations of 1 and 10 microg/ml. However, at a concentration of 100 microg/ml, ITS was able to protect sperm during short-term hypothermic storage. Addition of AFP or ITS at 100 microg/ml to test egg yolk-glycerol extender during freezing significantly (P < 0.05) increased postthaw motility, plasma membrane integrity, and acrosome integrity. The mean (+/-SE) motility recovery rate increased from 28.9 +/- 3.9%, for the untreated control, to 59.2 +/- 5.8% and 67.8 +/- 7.4%, for ITS and AFP, respectively. The effects of the thawing procedure were influenced by the presence of AFP during the freezing cycle. An improved motility recovery rate of 67 +/- 4.2% was obtained when chimpanzee sperm frozen in test egg yolk-glycerol extender supplemented with AFP were thawed rapidly at 37 degrees C, compared to 47 +/- 5.2% and 44 +/- 8.2% for slow (23 degrees C) and ultrarapid (75 degrees C) thawing, respectively. The motility recovery after thawing of ITS-treated semen at 23 degrees C, 37 degrees C, or 75 degrees C was not significantly different. Semen frozen without AFP or ITS and thawed at 75 degrees C was seriously (P < 0.05) damaged. This study provides evidence that AFP- or ITS-supplemented semen extender improves postthaw sperm motility in the chimpanzee.
我们研究了抗冻肽（AFP）和胰岛素转铁蛋白硒（ITS）对黑猩猩（Pan troglodytes）精子在冷冻（0 - 5摄氏度）和解冻后活力及膜完整性的影响。还检测了三种解冻程序在有无AFP和ITS存在的情况下，对精子活力以及质膜和顶体状态的影响。在冷冻过程中，AFP和ITS似乎具有轻度细胞毒性，因为当AFP浓度为1、10和100微克/毫升以及ITS浓度为1和10微克/毫升时，精子的前向运动能力和速度（曲线速度和直线速度）显著下降。然而，在浓度为100微克/毫升时，ITS能够在短期低温保存期间保护精子。在冷冻过程中向测试卵黄 - 甘油稀释液中添加100微克/毫升的AFP或ITS，显著（P < 0.05）提高了解冻后的精子活力、质膜完整性和顶体完整性。平均（±标准误）活力恢复率从未经处理的对照组的28.9 ± 3.9%，分别提高到ITS组的59.2 ± 5.8%和AFP组的67.8 ± 7.4%。解冻程序的效果受冷冻周期中AFP存在的影响。当在添加AFP的测试卵黄 - 甘油稀释液中冷冻的黑猩猩精子在37摄氏度快速解冻时，活力恢复率提高到67 ± 4.2%，相比之下，缓慢（23摄氏度）解冻和超快速（75摄氏度）解冻的恢复率分别为47 ± 5.2%和44 ± 8.2%。ITS处理的精液在23摄氏度、37摄氏度或75摄氏度解冻后的活力恢复没有显著差异。未添加AFP或ITS冷冻并在75摄氏度解冻的精液受到严重（P < 0.05）损伤。本研究提供了证据表明，添加AFP或ITS的精液稀释液可提高黑猩猩精子解冻后的活力。
