Holden C A, Trounson A O
Centre for Early Human Development, Monash Medical Centre, Victoria, Australia.
J Exp Zool. 1992 Dec 15;264(4):468-74. doi: 10.1002/jez.1402640413.
The present study investigates whether a 5 hour capacitation period modifies the ability of human spermatozoa to undergo induced acrosomal loss. Human sperm acrosomal loss was induced by treatment with either the calcium ionophore A23187, low concentrations of the phospholipid dilauroylphosphatidylcholine (PC12), or 2 hours incubation in conditioned medium prepared from human cumulus cells (CM/CC). The use of a dual staining method (FITC-ConA and Hoechst 33258) for simultaneous assessment of acrosomal status and viability demonstrated that induction of acrosomal loss with calcium ionophore was not dependent on a capacitation period. A short (5 hour) incubation period was not sufficient to induce acrosomal loss with CM/CC above spontaneous acrosome reaction rates in medium alone. A significant capacitation-dependent increase (P < 0.05) in acrosomal loss was observed when human spermatozoa were incubated with PC12. Induction of acrosomal loss of capacitated human spermatozoa with PC12 therefore provides a simple assay for the simultaneous assessment of human sperm capacitation and the acrosome reaction in vitro.
本研究调查了5小时的获能期是否会改变人类精子发生诱导顶体丢失的能力。通过用钙离子载体A23187、低浓度的磷脂二月桂酰磷脂酰胆碱（PC12）处理，或在由人卵丘细胞制备的条件培养基（CM/CC）中孵育2小时来诱导人类精子顶体丢失。使用双重染色方法（FITC-ConA和Hoechst 33258）同时评估顶体状态和活力表明，用钙离子载体诱导顶体丢失不依赖于获能期。短（5小时）孵育期不足以在仅培养基中高于自发顶体反应率的情况下，用CM/CC诱导顶体丢失。当人类精子与PC12孵育时，观察到顶体丢失有显著的获能依赖性增加（P < 0.05）。因此，用PC12诱导获能的人类精子顶体丢失为体外同时评估人类精子获能和顶体反应提供了一种简单的检测方法。
