Zitta Karina, Wertheimer Eva, Miranda Patricia V
Instituto de Biología y Medicina Experimental-CONICET, Buenos Aires, C1428ADN, Argentina.
Mol Hum Reprod. 2004 Dec;10(12):925-33. doi: 10.1093/molehr/gah122. Epub 2004 Oct 29.
Glycoproteins and lectin-like proteins mediate sperm-zona pellucida interaction. The present study analysed the participation of carbohydrates in the different stages of sperm interaction with the zona pellucida in hamster, by determining the effects of different monosaccharides. N-acetylglucosamine (GlcNAc, 1 mM) reduced sperm ability to bind to the zona pellucida. Surprisingly, spontaneous acrosome reaction (AR) was also inhibited by this sugar. In order to analyse the effect of GlcNAc on sperm-zona pellucida binding, independent of its effect on the AR, strontium (Sr) was used as a calcium (Ca) replacement in the sperm capacitation and co-incubation medium. Sr seemed to be able to replace Ca for sperm capacitation, at least when measured as the ability to bind to the zona pellucida, and undergo AR when Ca is provided. Moreover, sperm-zona pellucida binding could also take place in a Sr-modified medium. When binding assays were carried out in the Sr medium, GlcNAc also produced an inhibitory effect. This could be reproduced when sperm, but not oocytes, were pre-incubated with the monosaccharide. IVF assays were also carried out to analyse the participation of GlcNAc in the different steps of sperm-oocyte interaction. Taken together, the results support the involvement of the GlcNAc residues of the zona pellucida in the early steps of the interaction with sperm.
糖蛋白和凝集素样蛋白介导精子与透明带的相互作用。本研究通过测定不同单糖的作用,分析了碳水化合物在仓鼠精子与透明带相互作用不同阶段的参与情况。N-乙酰葡糖胺(GlcNAc,1 mM)降低了精子与透明带结合的能力。令人惊讶的是,这种糖还抑制了自发顶体反应(AR)。为了分析GlcNAc对精子-透明带结合的影响,而不考虑其对AR的影响,在精子获能和共孵育培养基中使用锶(Sr)替代钙(Ca)。Sr似乎能够替代Ca进行精子获能,至少以与透明带结合的能力来衡量,并且在提供Ca时能发生AR。此外,精子-透明带结合也能在Sr改良培养基中发生。当在Sr培养基中进行结合试验时,GlcNAc也产生抑制作用。当精子而非卵母细胞与单糖预孵育时,这种抑制作用可以重现。还进行了体外受精试验以分析GlcNAc在精子-卵母细胞相互作用不同步骤中的参与情况。综上所述,结果支持透明带的GlcNAc残基参与了与精子相互作用的早期步骤。