Gundersen G G, Shapiro B M
J Cell Biol. 1984 Oct;99(4 Pt 1):1343-53. doi: 10.1083/jcb.99.4.1343.
Certain sperm components labeled with fluorescein isothiocyanate or its radioactive derivative, 125I-diiodofluorescein isothiocyanate (125IFC), are transferred at fertilization to the egg, where they persist throughout early cleavage stages at a localized site in the embryo cytoplasm (Gabel, C. A., E. M. Eddy, and B. M. Shapiro, 1979, Cell, 18:207-215; Gundersen, G. G., C. A. Gabel, and B. M. Shapiro, 1982, Dev. Biol., 93:59-72). By using image intensification we have extended these observations in the sea urchin to the pluteus larval stage, in which greater than 60% of the embryos have localized fluorescent sperm components. Because of the unusual persistence of the sperm components in the embryo, a characterization of the nature of the labeled species in sea urchin sperm was undertaken. Approximately 10% of the 125IFC was in sperm polypeptides of Mr greater than 15,000. These proteins were on the sperm surface as shown by their sensitivity to externally added proteases. The remainder of the 125IFC in sperm was in several low-molecular-weight species, none of which was 125IFC-derivatized phospholipid. To determine if any labeled sperm polypeptides remained intact in the embryo after fertilization, 125IFC-labeled sperm proteins were recovered from one-cell and late gastrula stage embryos by using an anti-IFC immunoadsorbent. Most of the labeled sperm proteins were degraded shortly after fertilization; however, distinct sets of labeled polypeptides were recovered from both one-cell and gastrula stage embryos. Six of the labeled polypeptides recovered from both embryonic stages had identical SDS gel mobilities as labeled sperm polypeptides. Other polypeptides in the embryos appeared to arise from limited proteolysis of sperm proteins. Thus, in this physiological cell fusion system, individual sperm proteins are transferred to the egg at fertilization, and some persist intact or after specific, limited degradation long after gamete fusion, until at least the late gastrula stage.
某些用异硫氰酸荧光素或其放射性衍生物125I - 二碘异硫氰酸荧光素（125IFC）标记的精子成分，在受精时会转移到卵子中，并在胚胎细胞质的一个局部位点在早期卵裂阶段一直存在（加贝尔，C.A.，E.M.埃迪，和B.M.夏皮罗，1979年，《细胞》，18:207 - 215；冈德森，G.G.，C.A.加贝尔，和B.M.夏皮罗，1982年，《发育生物学》，93:59 - 72）。通过使用图像增强技术，我们将海胆中的这些观察结果扩展到了长腕幼虫阶段，其中超过60%的胚胎有局部荧光精子成分。由于精子成分在胚胎中异常持久，因此对海胆精子中标记物种的性质进行了表征。125IFC的大约10%存在于分子量大于15,000的精子多肽中。这些蛋白质位于精子表面，这可通过它们对外部添加蛋白酶的敏感性来证明。精子中其余的125IFC存在于几种低分子量物种中，其中没有一种是125IFC衍生的磷脂。为了确定受精后胚胎中是否有任何标记的精子多肽保持完整，通过使用抗IFC免疫吸附剂从单细胞和原肠胚晚期胚胎中回收了125IFC标记的精子蛋白。大多数标记的精子蛋白在受精后不久就被降解；然而，从单细胞和原肠胚阶段胚胎中都回收了不同的标记多肽组。从两个胚胎阶段回收的六个标记多肽与标记的精子多肽具有相同的SDS凝胶迁移率。胚胎中的其他多肽似乎源于精子蛋白的有限蛋白水解。因此，在这个生理细胞融合系统中，单个精子蛋白在受精时转移到卵子中，并且一些在配子融合后很长时间保持完整或经过特定的有限降解后仍然存在，直到至少原肠胚晚期。
