Servetnick M D, Wilt F H
Dev Biol. 1987 Sep;123(1):231-44. doi: 10.1016/0012-1606(87)90445-3.
A rapid, gentle technique is described for the isolation of nuclei from sea urchin embryos. Using this technique, we have analyzed the synthesis and accumulation of nonhistone nuclear proteins during sea urchin development by two-dimensional gel electrophoresis. Most nuclear proteins fall into one of three patterns of synthesis, which are distinguished by maximal rates of accumulation at early (prior to hatching blastula), middle (hatching blastula/gastrula), or late (prism/pluteus) stages of development. Over 60% of observed nuclear proteins undergo apparent qualitative changes in synthesis and accumulation between the 64-cell and pluteus stages. Most of these changes represent appearances of new proteins. A large number of qualitative changes occur very early in development; the period of greatest change is between the 64-cell and 200-cell stages. Over half of the proteins which first appear in the nucleus subsequent to the 64-cell stage are synthesized at stages prior to the time of their initial appearance in nuclei, but are excluded from nuclei for some time.
本文描述了一种从海胆胚胎中分离细胞核的快速、温和技术。利用该技术，我们通过二维凝胶电泳分析了海胆发育过程中非组蛋白核蛋白的合成与积累。大多数核蛋白的合成模式可分为三种，其区别在于在发育早期（囊胚孵化前）、中期（囊胚孵化/原肠胚期）或晚期（棱柱幼虫/长腕幼虫期）的最大积累速率。超过60%的观察到的核蛋白在64细胞期和长腕幼虫期之间的合成与积累过程中经历了明显的定性变化。这些变化大多表现为新蛋白的出现。大量的定性变化在发育早期就发生了；变化最大的时期是在64细胞期和200细胞期之间。在64细胞期之后首次出现在细胞核中的蛋白质中，超过一半是在它们最初出现在细胞核之前的阶段合成的，但在一段时间内被排除在细胞核之外。
