Jäckle Herbert, Kalthoff Klaus
Department of Zoology, University of Texas at Austin, 78712, Austin, Texas, USA.
Wilehm Roux Arch Dev Biol. 1979 Dec;187(4):283-305. doi: 10.1007/BF00848465.
Embryos of the chironomid midgeSmittia spec. were permeabilized with sodium hypochlorite and octane. Uptake of labeled uridine and amino acids suggested that these compounds are actively transported across the plasma membrane. Before blastoderm formation, uridine was incorporated at low levels into nuclear DNA and mitochondrial RNA. After blastoderm formation, uridine was incorporated rapidly, mostly into cytoplasmic RNA including both ribosomal RNA precursors and poly(A)-containing RNA. Protein synthesis was observed throughout early embryogenesis. By measuring incorporation of labeled amino acids and internal amino acid pool sizes, we found that the rate of protein synthesis increased with development. Experiments with inhibitors of transcription indicated that proteins synthesized before blastoderm formation were translated from maternal mRNA. During blastoderm stages, embryonic mRNAs seemed to replace maternal mRNAs. Proteins synthesized during short incubation periods in vivo were separated by two-dimensional gel electrophoresis. After blastoderm formation, several new proteins were found that could not be detected at earlier stages.
摇蚊属的斯氏摇蚊胚胎用次氯酸钠和辛烷进行通透处理。标记尿苷和氨基酸的摄取表明这些化合物是通过质膜主动运输的。在胚盘形成之前,尿苷以低水平掺入核DNA和线粒体RNA中。胚盘形成后,尿苷迅速掺入,主要掺入细胞质RNA中,包括核糖体RNA前体和含poly(A)的RNA。在整个早期胚胎发育过程中都观察到了蛋白质合成。通过测量标记氨基酸的掺入和内部氨基酸池大小,我们发现蛋白质合成速率随着发育而增加。转录抑制剂实验表明,胚盘形成前合成的蛋白质是由母体mRNA翻译而来的。在胚盘阶段,胚胎mRNA似乎取代了母体mRNA。体内短时间孵育期间合成的蛋白质通过二维凝胶电泳进行分离。胚盘形成后,发现了几种在早期阶段无法检测到的新蛋白质。