Ahnonkitpanit V, White D, Suwajanakorn S, Kan F W, Namking M, Wells G, Kamolvarin N, Tanphaichitr N
Human IVF Program, Reproductive Biology Unit, Hormones/Growth/Development Research Group, Loeb Research Institute, Ottawa Civic Hospital, Department of Biochemistry, University of Ottawa, Ontario, Canada.
Biol Reprod. 1999 Sep;61(3):749-56. doi: 10.1095/biolreprod61.3.749.
We have shown that sperm sulfolipidimmobilizing protein 1 (SLIP1, molecular mass of 68 kDa), a sulfogalactosylglycerolipid (SGG)-binding protein, is significant in sperm-zona pellucida (ZP) interaction. The objective of this study was to localize SLIP1 on the egg and determine its role in gamete interaction. Immunofluorescence and immunoprotein A gold electron microscopy localized SLIP1 to the egg plasma membrane. In vitro gamete binding, using zona-free eggs preincubated with antiSLIP1 Fab before coincubation with sperm, showed a significant, dose-dependent decrease in sperm-egg plasma membrane binding. Similar results were obtained when affinity-purified antiSLIP1 IgG was used for egg pretreatment. The significance of egg SLIP1 in sperm-egg plasma membrane binding was further demonstrated by a decrease (36-52%) in in vitro fertilization when zona-intact eggs were pretreated with antiSLIP1 IgG. Since SLIP1 has been shown to bind SGG in vitro, we investigated the possibility that sperm SGG may participate in sperm-egg plasma membrane binding through egg SLIP1. Pretreatment of sperm with antiSGG Fab prior to coincubation with zona-free eggs resulted in a dose-dependent decrease in sperm-egg plasma membrane binding. Collectively, these findings strongly suggest a role for egg SLIP1 in sperm-egg plasma membrane interaction, which may be through its binding to sperm SGG.
我们已经表明,精子硫脂固定蛋白1(SLIP1,分子量为68 kDa),一种磺基半乳糖基甘油脂(SGG)结合蛋白,在精子-透明带(ZP)相互作用中具有重要意义。本研究的目的是将SLIP1定位在卵子上,并确定其在配子相互作用中的作用。免疫荧光和免疫蛋白A金电子显微镜将SLIP1定位在卵子质膜上。在与精子共孵育之前,使用预先用抗SLIP1 Fab孵育的无透明带卵子进行体外配子结合实验,结果显示精子-卵子质膜结合显著且呈剂量依赖性降低。当使用亲和纯化的抗SLIP1 IgG对卵子进行预处理时,也获得了类似的结果。当用抗SLIP1 IgG对完整透明带的卵子进行预处理时,体外受精率降低了(36 - 52%),这进一步证明了卵子SLIP1在精子-卵子质膜结合中的重要性。由于SLIP1已被证明在体外能结合SGG,我们研究了精子SGG可能通过卵子SLIP1参与精子-卵子质膜结合的可能性。在与无透明带卵子共孵育之前,用抗SGG Fab对精子进行预处理,导致精子-卵子质膜结合呈剂量依赖性降低。总的来说,这些发现有力地表明卵子SLIP1在精子-卵子质膜相互作用中发挥作用,这可能是通过其与精子SGG的结合来实现的。