Olson J H, Chandler D E
Department of Biology and the Program in Molecular and Cellular Biology, Arizona State University, Tempe, Arizona, 85287-1501, USA.
Dev Biol. 1999 Jun 15;210(2):401-10. doi: 10.1006/dbio.1999.9281.
The eggs of Xenopus laevis are surrounded by investment layers of egg jelly that interact with the sperm immediately prior to fertilization. Components of these egg jelly layers are necessary for the fertilization of the egg by incoming sperm. Eggs which are stripped of their jelly layers are refractile to fertilization by sperm, but the addition of solubilized jelly promotes fertilization. We have shown previously that the egg jelly layers are composed of a fibrous network of glycoconjugates which loosely hold smaller diffusible components. Extracts of these diffusible components were prepared by incubation of freshly ovulated eggs in high-salt buffers for 12 h at 4 degrees C. This diffusible component extract, when incubated with sperm, promoted the sperm's ability to fertilize dejellied eggs in a dose-dependent manner. In contrast, the high-molecular-weight "structural" glycoconjugates of jelly that remain after extraction of the diffusible components did not increase fertilization efficiency of dejellied eggs nor did nonspecific proteins, carbohydrate polymers, or organic polymers. The diffusible components, analyzed by SDS-PAGE, consisted of a mixture of proteins from 4 to 180 kDa. The protein responsible for fertilization rescue appeared to be <50 kDa and appeared to self-aggregate or to bind to larger proteins. This protein component was required during sperm binding to the egg, its action required an intact egg vitelline envelope, and its action was independent of large soluble polymers such as Ficoll.
非洲爪蟾的卵被卵胶的包被层所环绕,这些包被层在受精前会与精子相互作用。这些卵胶层的成分对于精子使卵受精是必需的。去除卵胶层的卵对精子受精具有抗性,但添加溶解的卵胶可促进受精。我们之前已经表明,卵胶层由糖缀合物的纤维网络组成,该网络松散地结合着较小的可扩散成分。通过将刚排卵的卵在4℃的高盐缓冲液中孵育12小时来制备这些可扩散成分的提取物。这种可扩散成分提取物与精子一起孵育时,以剂量依赖的方式促进精子使去胶卵受精的能力。相比之下,在提取可扩散成分后残留的卵胶的高分子量“结构”糖缀合物、非特异性蛋白质、碳水化合物聚合物或有机聚合物均未提高去胶卵的受精效率。通过SDS-PAGE分析,可扩散成分由4至180 kDa的蛋白质混合物组成。负责受精拯救的蛋白质似乎小于50 kDa,并且似乎会自我聚集或与更大的蛋白质结合。在精子与卵结合过程中需要这种蛋白质成分,其作用需要完整的卵黄膜,并且其作用独立于诸如聚蔗糖等大型可溶性聚合物。