DasGupta S, Mills C L, Fraser L R
Diagnostic Andrology Service, Hallam Medical Centre, London, UK.
J Reprod Fertil. 1993 Sep;99(1):135-43. doi: 10.1530/jrf.0.0990135.
Chlortetracycline (CTC) fluorescence patterns were used to assess Ca(2+)-related changes in the capacitation state of human spermatozoa incubated under conditions that would affect their intracellular Ca2+ levels. Initial experiments were designed to identify consistently occurring patterns and to correlate these with acrosomal status. Incubation for up to 1 h with the ionophore A23187 (10 mumol l-1), known to promote capacitation and acrosomal exocytosis, allowed the identification of three different CTC staining patterns which were very similar to those described for mouse spermatozoa. For this reason, they were given the same nomenclature: 'F'-characteristic of uncapacitated, acrosome-intact cells; 'B'-characteristic of capacitated, acrosome-intact cells; and 'AR'-characteristic of capacitated, acrosome-reacted cells. The distribution of the three patterns in the ionophore-treated suspensions was very different from that in control suspensions treated with dimethylsulfoxide only, with a significantly higher proportion of cells displaying the B and AR patterns and a significantly lower number of cells displaying the F pattern in the ionophore-treated group at all times. A strong concordance was found between the acrosomal status of cells determined using both CTC and fluorescein-conjugated Pisum sativum agglutinin (PSA) staining methods on the same cells. Verification of PSA staining patterns with acrosomal status was obtained by means of transmission electron microscopy. The proportion of cells with uniform fluorescence in the acrosomal region correlated with acrosome-intact cells; those with only equatorial segment staining correlated with fully-reacted cells, and those exhibiting equatorial fluorescence and patchy fluorescence over the rest of the acrosomal region correlated with cells in intermediate stages of exocytosis.(ABSTRACT TRUNCATED AT 250 WORDS)
金霉素（CTC）荧光模式被用于评估在会影响其细胞内钙离子水平的条件下孵育的人类精子获能状态中与钙离子相关的变化。最初的实验旨在识别持续出现的模式，并将这些模式与顶体状态相关联。用已知可促进获能和顶体胞吐作用的离子载体A23187（10 μmol l-1）孵育长达1小时，可识别出三种不同的CTC染色模式，这些模式与小鼠精子所描述的模式非常相似。因此，它们被赋予相同的命名：“F”——未获能、顶体完整细胞的特征；“B”——获能、顶体完整细胞的特征；“AR”——获能、顶体反应细胞的特征。离子载体处理的悬浮液中这三种模式的分布与仅用二甲基亚砜处理的对照悬浮液中的分布非常不同，在离子载体处理组中，始终有显著更高比例的细胞呈现B和AR模式，而呈现F模式的细胞数量显著更低。在对同一细胞使用CTC和荧光素偶联的豌豆凝集素（PSA）染色方法测定的细胞顶体状态之间发现了很强的一致性。通过透射电子显微镜对PSA染色模式与顶体状态进行了验证。顶体区域荧光均匀的细胞比例与顶体完整细胞相关；仅赤道段染色的细胞与完全反应的细胞相关，而在顶体区域其余部分呈现赤道荧光和斑片状荧光的细胞与胞吐作用中间阶段的细胞相关。（摘要截于250字）
