Otto J J, Kane R E, Bryan J
Cell Motil. 1980;1(1):31-40. doi: 10.1002/cm.970010104.
Following fertilization, the sea urchin egg cortex undergoes a structural change involving the assembly and organization of actin filaments into microvilli. Antifascin localizes this actin cross-linking protein in the microvilli of the fertilized egg cortex but no organized staining is present in the unfertilized cortex. Determination of the actin content of eggs using the DNAase I inhibition assay indicates that actin is about 1.4% of the total protein. Approximately 90% of this actin is soluble in low calcium isotonic extracts of unfertilized eggs while only 60-65% can be recovered in identical extracts of fertilized eggs. Similar measurements for fascin using a radioimmunoassay indicate this molecule represents about 0.3% of the total egg protein, essentially all of which is recovered in low calcium isotonic extracts of unfertilized eggs. After fertilization only 65-70% of this actin cross-linking protein is in the soluble phase. These results demonstrate a markedly different solubility for actin and fascin after fertilization, when the indirect immunofluorescence staining localizes fascin in the microvilli, and are consistent with the idea that fascin organizes newly polymerized actin filaments into the microvillar cores. A consideration of the amounts of actin and fascin incorporated into the cortex after fertilization and the number of microvilli on the egg surface indicates that the measured values are sufficient to account for the observed microvillar elongation.
受精后,海胆卵皮质会发生结构变化,涉及肌动蛋白丝组装并组织成微绒毛。抗肌动蛋白纤丝蛋白将这种肌动蛋白交联蛋白定位在受精卵皮质的微绒毛中,但未受精的皮质中不存在有组织的染色。使用DNA酶I抑制试验测定卵中的肌动蛋白含量表明,肌动蛋白约占总蛋白的1.4%。这种肌动蛋白中约90%可溶于未受精卵的低钙等渗提取物中,而在受精卵的相同提取物中只能回收60 - 65%。使用放射免疫测定法对肌动蛋白纤丝蛋白进行的类似测量表明,该分子约占卵总蛋白的0.3%,基本上所有该分子都可从未受精卵的低钙等渗提取物中回收。受精后,这种肌动蛋白交联蛋白只有65 - 70%处于可溶阶段。这些结果表明,受精后肌动蛋白和肌动蛋白纤丝蛋白的溶解度明显不同,此时间接免疫荧光染色将肌动蛋白纤丝蛋白定位在微绒毛中,这与肌动蛋白纤丝蛋白将新聚合的肌动蛋白丝组织成微绒毛核心的观点一致。考虑到受精后掺入皮质的肌动蛋白和肌动蛋白纤丝蛋白的量以及卵表面微绒毛的数量表明,测量值足以解释观察到的微绒毛伸长。