Robertson L, Wolf D P, Tash J S
Department of Cell Biology, Baylor College of Medicine, Texas Medical Center, Houston 77030.
Biol Reprod. 1988 Nov;39(4):797-805. doi: 10.1095/biolreprod39.4.797.
The occurence and time course of capacitation, acrosomal loss, and hyperactivated motility require quantitative definition in order to characterize fertile human sperm. In this study, video microscopy and digital image analysis were used to measure curvilinear (VCL) and straight line (VSL) velocity, average linearity of progression (LIN [100 x VSL/VCL]), maximum and mean amplitude of lateral head displacement (ALH), beat-cross-frequency (BCF), DANCE (VCL x meanALH) and DANCEMEAN (meanALH/(LIN/100]. These parameters were measured for sperm in semen and in the swim-up fraction of washed cells during incubation for up to 24 h under in vitro fertilization (IVF) conditions. Acrosomal loss was monitored in the same population of washed cells by an immunofluorescence end-point assay. The greatest increase in mean values of motility parameters was observed when seminal sperm were washed free of seminal plasma. Increases continued for up to 6 h of incubation. Two subpopulations of hyperactivated sperm were identified; one type, not found in semen, showed star-spin trajectories, and constituted 3.0, 3.8, 4.5, and 4.1% of the swim-up population after 0, 3, 6 and 24 h of incubation. The second type, termed transitional showed a more progressive trajectory and constituted less than 1% in semen. In total, hyperactivated cells constituted 0.8% of cells in semen, 14.5% of the swim-up population with no incubation, and 23.1, 22.7, and 19.4% after 3, 6, and 24 h of incubation, respectively. Acrosomal loss in the swim-up population was delayed during the first 3 h of incubation, then increased from near 5% at 3 h to 7 and 12% at 6 and 24 h, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
为了表征具有生育能力的人类精子，需要对获能、顶体丢失和超活化运动的发生及时间进程进行定量定义。在本研究中，利用视频显微镜和数字图像分析来测量曲线速度（VCL）和直线速度（VSL）、平均前进线性度（LIN [100×VSL/VCL]）、头部横向位移的最大和平均幅度（ALH）、鞭打交叉频率（BCF）、DANCE（VCL×平均ALH）以及DANCEMEAN（平均ALH/（LIN/100））。在体外受精（IVF）条件下，对精液中的精子以及洗涤后的细胞上浮部分在长达24小时的孵育过程中测量这些参数。通过免疫荧光终点测定法对同一批洗涤后的细胞群体监测顶体丢失情况。当精液精子洗去精浆后，观察到运动参数平均值的最大增加。增加持续至孵育6小时。鉴定出两种超活化精子亚群；一种在精液中未发现，呈现星状旋转轨迹，在孵育0、3、6和24小时后分别占上浮群体的3.0%、3.8%、4.5%和4.1%。第二种类型称为过渡型，呈现更具前进性的轨迹，在精液中占比不到1%。总体而言，超活化细胞在精液中占细胞总数的0.8%，未孵育的上浮群体中占14.5%，在孵育3、6和24小时后分别占23.1%、22.7%和19.4%。上浮群体中的顶体丢失在孵育的前3小时延迟，然后从3小时时的近5%分别增加到6小时时的7%和24小时时的12%。（摘要截短至250字）
