Livingston B T, Wilt F H
Department of Zoology, University of California, Berkeley 94720.
Proc Natl Acad Sci U S A. 1989 May;86(10):3669-73. doi: 10.1073/pnas.86.10.3669.
The mechanism of determination of early embryonic cells has been investigated using sea urchin embryos. An efficacious method of isolating blastomere pairs from the animal or vegetal half of sea urchin embryos was developed. The overt differentiation of separated animal and vegetal blastomere pairs resembles that of separated animal and vegetal hemispheres isolated by manual dissection. Treatment of animal blastomeres with LiCl caused them to display a morphology resembling that of isolated vegetal blastomeres. The effects of separation of animal and vegetal blastomeres and of treatment of animal blastomeres with LiCl were examined at the molecular level using gut alkaline phosphatase and a spicule matrix protein RNA as markers of differentiation. Histochemical staining and in situ hybridization studies showed that these markers are normally only expressed in vegetal blastomeres but that their expression can be evoked in animal blastomeres by treatment with LiCl.
利用海胆胚胎对早期胚胎细胞的决定机制进行了研究。开发了一种从海胆胚胎的动物极或植物极分离卵裂球对的有效方法。分离的动物极和植物极卵裂球对的明显分化类似于通过手工解剖分离的动物极和植物极半球的分化。用LiCl处理动物极卵裂球会使其呈现出类似于分离的植物极卵裂球的形态。使用肠道碱性磷酸酶和一种骨针基质蛋白RNA作为分化标记,在分子水平上研究了动物极和植物极卵裂球分离以及用LiCl处理动物极卵裂球的效果。组织化学染色和原位杂交研究表明,这些标记通常仅在植物极卵裂球中表达,但通过用LiCl处理可在动物极卵裂球中诱导其表达。