Fukuda M, Morales P, Overstreet J W
Department of Obstetrics and Gynecology, School of Medicine, University of California, Davis.
Gamete Res. 1989 Sep;24(1):59-65. doi: 10.1002/mrd.1120240109.
In previous studies we have shown differences in the function of morphologically normal and abnormal sperm by evaluating their flagellar movements and swimming trajectories. In this study we have compared the capability of morphologically normal and abnormal human sperm to undergo an acrosome reaction after incubation with human follicular fluid. Semen samples were studied from 6 research donors and 21 semen evaluation patients. All men had normal semen by clinical criteria. Semen was prepared either by a two-step Percoll gradient centrifugation or the sperm were diluted, washed, and centrifuged three times. Sperm suspensions were incubated for 24 hours in a modified Tyrode's medium, containing 2.6% bovine serum albumin, prior to dilution with human follicular fluid. The percentage of acrosome reactions among viable sperm was assessed after 15 minutes using the supravital Hoescht stain and fluoresceinated pea lectin. Sperm head size was measured with an ocular micrometer and normal values were defined as length 3-5 microns and width 2-3 microns. At least 25 viable normal sperm, and 25 viable abnormal sperm were analyzed for acrosome reactions on each slide. With Percoll separation the percentage of acrosome reactions (mean +/- sem) for normal sperm was 38 +/- 3% vs. 22 +/- 2% for abnormal sperm (P less than 0.005). After washing, the comparable values were 12 +/- 1% vs. 5 +/- 1% (P less than 0.005). The incidence of spontaneous acrosome reactions (24 hours of incubation, no follicular fluid) was also higher for normal sperm than abnormal sperm (9 +/- 1% vs. 4 +/- 1%, P less than 0.01). These data demonstrate an association between normal sperm morphology and acrosomal function.
在之前的研究中,我们通过评估形态正常和异常精子的鞭毛运动及游动轨迹,展示了它们在功能上的差异。在本研究中,我们比较了形态正常和异常的人类精子在与人卵泡液孵育后发生顶体反应的能力。对6名研究捐赠者和21名精液评估患者的精液样本进行了研究。根据临床标准,所有男性的精液均正常。精液要么通过两步法Percoll梯度离心法制备,要么将精子稀释、洗涤并离心三次。在用人卵泡液稀释之前,精子悬液在含有2.6%牛血清白蛋白的改良Tyrode培养基中孵育24小时。15分钟后,使用活体染料Hoescht染色和荧光素化豌豆凝集素评估活精子中顶体反应的百分比。用目镜测微计测量精子头部大小,正常值定义为长度3 - 5微米,宽度2 - 3微米。每张载玻片上至少分析25个活的正常精子和25个活的异常精子的顶体反应。通过Percoll分离法,正常精子顶体反应的百分比(平均值±标准误)为38±3%,异常精子为22±2%(P<0.005)。洗涤后,相应的值分别为12±1%和5±1%(P<0.005)。正常精子自发顶体反应的发生率(孵育24小时,无卵泡液)也高于异常精子(9±1%对4±1%,P<0.01)。这些数据表明正常精子形态与顶体功能之间存在关联。