Alexandraki D, Ruderman J V
Dev Biol. 1985 Jun;109(2):436-51. doi: 10.1016/0012-1606(85)90470-1.
Mature unfertilized eggs of the sea urchin Lytechinus pictus contain multiple alpha-tubulin mRNAs, which range in size from 1.75 to 4.8 kb, and two beta-tubulin mRNAs, 1.8 and 2.25 kb. These mRNAs were found at similar levels throughout the early cleavage stages. RNA gel blot hybridizations showed that prominent quantitative and qualitative changes in tubulin mRNAs occurred between the early blastula and hatched blastula stages. The overall amounts of alpha- and beta-tubulin mRNAs increased two- to fivefold between blastula and pluteus. These increases were due mainly to a rise in a 1.75-kb alpha RNA and a new 2.0-kb beta RNA. Other, minor changes also occurred during subsequent development. All size classes of alpha- and beta-tubulin RNAs in early and late embryos contained poly(A)+ translatable sequences. As reported earlier, some of each of the alpha RNAs, but neither of the beta RNAs, are translated in the egg and a small portion of each of the stored alpha and beta RNAs is recruited onto polysomes within 30 min of fertilization. In the work described here, subsequent development up to the morula stage was accompanied by a gradual recruitment of tubulin mRNAs into polysomes. By the early blastula stage, most of the maternal tubulin sequences were associated with polysomes. In contrast to the gradual recruitment of maternal sequences throughout cleavage, the tubulin mRNAs which appeared at the blastula stage showed no delay in entering polysomes. The exact fraction of each mRNA that was translationally active at later stages varied somewhat among the individual mRNAs. From the differential hybridization patterns of egg, embryo, and testis RNAs to various tubulin cDNA and genomic DNA probes, it is concluded that at least one gene producing maternal alpha mRNA is different from a second one which is expressed only in testis. Each of the three embryonic beta RNAs is encoded by a different beta gene; at least two of these different beta genes are also expressed in testis.
太平洋紫海胆成熟未受精卵含有多种α-微管蛋白mRNA,大小在1.75至4.8 kb之间,以及两种β-微管蛋白mRNA,分别为1.8 kb和2.25 kb。在早期卵裂阶段,这些mRNA的水平相似。RNA凝胶印迹杂交显示,在囊胚早期和孵化囊胚阶段之间,微管蛋白mRNA发生了显著的数量和质量变化。从囊胚到长腕幼虫阶段,α-和β-微管蛋白mRNA的总量增加了两到五倍。这些增加主要是由于1.75 kb的αRNA和新出现的2.0 kb的βRNA的增加。在随后的发育过程中也发生了其他一些微小变化。早期和晚期胚胎中所有大小类别的α-和β-微管蛋白RNA都含有聚腺苷酸+可翻译序列。如先前报道,每种αRNA中的一些,但两种βRNA都没有在卵中翻译,并且在受精后30分钟内,储存的α-和β-微管蛋白RNA中的一小部分被募集到多核糖体上。在本文所述的工作中,直到桑椹胚阶段的后续发育伴随着微管蛋白mRNA逐渐募集到多核糖体中:到囊胚早期,大多数母体微管蛋白序列与多核糖体相关。与整个卵裂过程中母体序列逐渐募集不同,在囊胚阶段出现的微管蛋白mRNA在进入多核糖体时没有延迟。在后期,每种mRNA具有翻译活性的确切比例在各个mRNA之间略有不同。从卵、胚胎和睾丸RNA与各种微管蛋白cDNA和基因组DNA探针的差异杂交模式可以得出结论:至少一个产生母体αmRNA的基因与另一个仅在睾丸中表达的基因不同。三种胚胎βRNA中的每一种都由不同的β基因编码;这些不同的β基因中至少有两个也在睾丸中表达。