Tanphaichitr N, Smith J, Mongkolsirikieart S, Gradil C, Lingwood C A
Department of Obstetrics and Gynecology, Loeb Research Institute, Ottawa Civic Hospital, Ontario, Canada.
Dev Biol. 1993 Mar;156(1):164-75. doi: 10.1006/dbio.1993.1067.
A sulfoglycolipid immobilizing protein, termed SLIP1, is a conserved germ cell membrane protein that has been shown in vitro to bind specifically to the mammalian germ-cell-specific sulfoglycolipid, sulfogalactosylglycerolipid (SGG). SLIP1 was extracted from the mouse sperm surface by a sucrose/ATP/EDTA solution. Indirect immunofluorescence and immunoprotein A-gold labeling of live mouse sperm indicate that SLIP1 was present on the acrosomal plasma membrane. Inclusion of low concentrations of exogenous purified SLIP1 in the in vitro mouse sperm-egg binding assay culture significantly decreased the number of sperm bound per egg. Inhibition was heat labile. SLIP1 on sperm, rather than that on eggs, appeared to be important for the binding process since preexposure of sperm to anti-SLIP1 decreased sperm binding to the eggs. Eggs preincubated with anti-SLIP1 were unaffected. Immunoblotting studies confirmed that SLIP1 was not present on the zona pellucida (ZP). SLIP1 binding ligand(s) on eggs were also involved in sperm-ZP binding since preincubation of eggs with exogenous SLIP1 before gamete coincubation significantly reduced the number of sperm bound per egg. The results suggest that SLIP1 is involved in mouse sperm-ZP binding.
一种名为SLIP1的固定化硫糖脂蛋白是一种保守的生殖细胞膜蛋白,体外实验表明它能特异性结合哺乳动物生殖细胞特异性硫糖脂——硫代半乳糖基甘油脂(SGG)。通过蔗糖/ATP/EDTA溶液从小鼠精子表面提取出SLIP1。对活小鼠精子进行间接免疫荧光和免疫蛋白A-金标记显示,SLIP1存在于顶体细胞膜上。在体外小鼠精子-卵子结合试验培养物中加入低浓度的外源性纯化SLIP1,显著降低了每个卵子结合的精子数量。这种抑制作用对热不稳定。精子上的SLIP1而非卵子上的SLIP1似乎对结合过程很重要,因为精子预先暴露于抗SLIP1会降低精子与卵子的结合。预先用抗SLIP1孵育的卵子不受影响。免疫印迹研究证实,透明带(ZP)上不存在SLIP1。卵子上的SLIP1结合配体也参与精子与ZP的结合,因为在配子共孵育前用外源性SLIP1预先孵育卵子,显著减少了每个卵子结合的精子数量。结果表明,SLIP1参与小鼠精子与ZP的结合。