APROTININ AND A SEMINAL PLASMA FACTOR INHIBIT THE MOTILITY OF DEMEMBRANATED REACTIVATED RABBIT SPERMATOZOA
APROTININ AND A SEMINAL PLASMA FACTOR INHIBIT THE MOTILITY OF DEMEMBRANATED REACTIVATED RABBIT SPERMATOZOA
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DOI：
10.1095/biolreprod28.4.788
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发表时间：
1983-01-01
影响因子：
3.6
通讯作者：
GAGNON, C
中科院分区：
文献类型：
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作者：
DELAMIRANDE, E;BARDIN, CW;GAGNON, C
对射精的兔精子的脱膜和再激活进行了研究。研究发现 ATP、Mg、谷氨酸、二硫苏糖醇 (DTT) 和 Tris-HCl 对于良好的再激活至关重要。通过这个实验模型，我们研究了蛋白酶抑制剂对 ATP 重新启动运动以及对已经活动的精子运动的影响。大豆胰蛋白酶抑制剂 (STI) 将重新激活的时间延长 7 至 8 倍，而胃酶抑素、抗凝血酶 III、苯甲基磺酰氟 (PMSF) 和 α-1-抗胰蛋白酶则没有显着效果。抑肽酶 (1.5μg/ml) 和亮抑酶肽 (50μg/ml) 完全阻止 ATP 重新启动运动；相同浓度的抑肽酶甚至可以阻断活动精子的运动。组织特异性精浆因子也可以阻止运动的重新启动和活动精子的运动。然而，停止运动所需的精浆量是阻止运动重新开始所需的精浆量的 2-3 倍。精浆中的抑制剂很可能不是蛋白酶或抑肽酶样蛋白酶抑制剂，因为精浆抑制剂的部分纯化制剂不会水解胰蛋白酶底物，不会被蛋白酶抑制剂抑制，并且没有显着的抑制胰蛋白酶的能力。数据表明，抑肽酶和精浆抑制剂通过不同的机制阻断运动。抑肽酶和精浆抑制剂代表了研究精子运动调节的两种新工具。
The demembranation and reactivation of ejaculated rabbit spermatozoa have been studied. ATP, Mg, glutamate, dithiothreitol (DTT), and Tris-HCl were found to be essential for a good reactivation. With this experimental model, we investigated the effects of protease inhibitors on the reinitiation of movement by ATP and on the movement of already motile spermatozoa. Soybean trypsin inhibitor (STI) prolonged the length of reactivation by 7- to 8-fold, whereas pepstatin, antithrombin III, phenylmethylsulfonyl fluoride (PMSF), and α-1-antitrypsin had no significant effect. Aprotinin (1.5µg/ml) and leupeptin (50µg/ml) completely prevented the reinitiation of movement by ATP; aprotinin at the same concentration even blocked the movement of motile spermatozoa.A tissue-specific seminal plasma factor could also prevent both the reinitiation of movement and the movement of motile spermatozoa. However, it took 2–3 times the amount of seminal plasma to stop the movement than to prevent the reinitiation of movement. The inhibitor in the seminal plasma is most probably not a protease or an aprotinin-like protease inhibitor since a partially purified preparation of the seminal plasma inhibitor does not hydrolyze a trypsin substrate, is not inhibited by protease inhibitors and has no significant capacity to inhibit trypsin.The data suggest that aprotinin and the seminal plasma inhibitor block movement through different mechanisms. Aprotinin and the seminal plasma inhibitor represent two new tools to study the regulation of sperm movement.