Graham J K, Foote R H, Parrish J J
Biol Reprod. 1986 Sep;35(2):413-24. doi: 10.1095/biolreprod35.2.413.
Incubation of bull sperm with liposomes made with phosphatidylcholine (PC) containing fatty acyl chains of either 10 (PC10) or 12 (PC12) carbons resulted in greater than 90% of the sperm exhibiting an acrosome reaction (AR) within 15 min. Liposomes of PC10 rapidly destroyed sperm motility while PC12 acrosome-reacted sperm remained motile for several h. Liposomes of PC with greater than or equal to 14-carbon fatty acyl chains had no effect on the AR or motility of sperm. The AR was not induced by lysophospholipids, because lysophospholipids were not detected in the PC liposomes, and the AR did not occur when lysophospholipids were tested at the same concentration as PC12. The concentration of PC12 necessary to induce maximal numbers of acrosome-reacted sperm varied with the concentration of sperm. The effect of PC12 on sperm also varied with the ratio of live to dead sperm in a sample. When 3 X 10(6) bull sperm/ml were treated with 0, 10, 20, and 30 microM PC12 for 7 min prior to addition to zona-free hamster eggs, 6, 6, 98, and 77% of the eggs were penetrated, respectively. Lipid concentrations of 0 microM and 10 microM did not affect the AR, whereas higher levels induced the AR in sperm. This procedure can quickly provide acrosome-reacted bull sperm for use with various in vitro fertilization procedures and for assessment of male fertility.
用含有10个(PC10)或12个(PC12)碳原子脂肪酰链的磷脂酰胆碱(PC)制成的脂质体与公牛精子孵育,结果超过90%的精子在15分钟内发生顶体反应(AR)。PC10脂质体迅速破坏精子活力,而PC12顶体反应后的精子仍能游动数小时。含有大于或等于14个碳原子脂肪酰链的PC脂质体对精子的AR或活力没有影响。AR不是由溶血磷脂诱导的,因为在PC脂质体中未检测到溶血磷脂,并且当以与PC12相同的浓度测试溶血磷脂时,未发生AR。诱导最大数量顶体反应精子所需的PC12浓度随精子浓度而变化。PC12对精子的作用也随样品中活精子与死精子的比例而变化。当每毫升3×10⁶个公牛精子在添加到无透明带仓鼠卵之前用0、10、20和30微摩尔PC12处理7分钟时,分别有6%、6%、98%和77%的卵被穿透。0微摩尔和10微摩尔的脂质浓度不影响AR,而较高水平可诱导精子发生AR。该方法可以快速提供顶体反应后的公牛精子,用于各种体外受精程序和评估男性生育能力。