Lynn D A, Angerer L M, Bruskin A M, Klein W H, Angerer R C
Proc Natl Acad Sci U S A. 1983 May;80(9):2656-60. doi: 10.1073/pnas.80.9.2656.
Spec 1 mRNAs increase 100-fold in abundance per embryo during early sea urchin development. Previous studies indicated an enrichment of this mRNA in ectoderm fractions of gastrulae and plutei. We have determined the precise localization of this mRNA by in situ hybridization techniques. In pluteus larvae, the mRNA is highly restricted to a set of morphologically uniform ectoderm cells in the dorsal part of the embryo. The mRNA is not detectable in other regions of ectoderm or in endoderm and mesoderm. The pattern of localization is already established at the gastrula stage, before these cells are distinguishable by morphological criteria. This pattern of distribution of Spec 1 mRNA is distinct from that of bulk poly(A)+ mRNA. Measurements of the amount of Spec 1 mRNA per embryo and the number of cells containing this RNA indicate that there are about 500 Spec 1 mRNA molecules per cell at the pluteus stage and probably twice as many at the gastrula stage. These results indicate that the sensitivity of the in situ hybridization method allows detection of sequences that comprise approximately equal to 0.05% of the embryo mRNA nucleotides.
在海胆早期发育过程中,Spec 1 mRNA在每个胚胎中的丰度增加了100倍。先前的研究表明,这种mRNA在原肠胚和长腕幼虫的外胚层部分中富集。我们已经通过原位杂交技术确定了这种mRNA的精确位置。在长腕幼虫中,mRNA高度局限于胚胎背部一组形态均匀的外胚层细胞中。在其他外胚层区域或内胚层和中胚层中检测不到这种mRNA。在原肠胚阶段,在这些细胞通过形态学标准可区分之前,定位模式就已经建立。Spec 1 mRNA的这种分布模式与大量多聚腺苷酸加尾(poly(A)+)mRNA的分布模式不同。对每个胚胎中Spec 1 mRNA的量以及含有这种RNA的细胞数量的测量表明,在长腕幼虫阶段每个细胞约有500个Spec 1 mRNA分子,在原肠胚阶段可能是其两倍。这些结果表明,原位杂交方法的灵敏度能够检测到约占胚胎mRNA核苷酸0.05%的序列。