Marikawa Y, Li Y, Elinson R P
Department of Zoology, University of Toronto, 25 Harbord Street, Toronto, M5S 3G5, Canada.
Dev Biol. 1997 Nov 1;191(1):69-79. doi: 10.1006/dbio.1997.8710.
The Xenopus egg contains maternal dorsal determinants that are specifically located at the vegetal cortex. To study physical and functional properties of the dorsal determinants, we took advantage of the animal-vegetal reversed embryo. The animal-vegetal reversed embryo is produced by inversion of the fertilized egg, which results in formation of ectoderm and endoderm from the unpigmented and the pigmented halves, respectively [Neff et al. (1983). Dev. Biol. 97, 103-112; Black and Gerhart (1985). Dev. Biol. 108, 310-324]. We demonstrated by cytoplasmic transplantation that the dorsal activity was specifically localized to the unpigmented cortical cytoplasm of the inverted egg, which is segregated into the future ectodermal lineage. This result suggests that the dorsal determinants are associated with the unpigmented cortex and are not dislodged by the inversion. In addition, we found that two vegetally localized transcripts, Xcat2 and Vg1 mRNAs, were present in the reversed animal pole of the inverted egg, suggesting their association with the unpigmented cortex. In order to compare the dorsal determinant activity with known dorsalizing molecules, we examined the expression pattern of Xnr3 and Siamois in the reversed embryo because these two genes are activated by the Wnt-pathway activators (Xwnt-8, beta-catenin, etc.) but not by other dorsalizing molecules (noggin, BVg1, etc.). Animal cap of the reversed embryo, which received the unpigmented cortex of the egg, expressed Xnr3 and Siamois. However, Mix.1, a marker expressed in endoderm and mesoderm in the normal embryo in response to mesodermal inducers, was not detected in the animal cap of the reversed embryo. In addition, we found that beta-catenin protein accumulated in nuclei of unpigmented animal pole cells of the reversed embryo. These results suggest that the maternal dorsal determinants behave more similarly to the Wnt-pathway activators than noggin or BVg1.
非洲爪蟾卵含有母体背侧决定因子,这些因子特异性地定位于植物性皮层。为了研究背侧决定因子的物理和功能特性,我们利用了动物 - 植物反转胚胎。动物 - 植物反转胚胎是通过受精卵的倒置产生的,这分别导致由未着色和着色的半部分形成外胚层和内胚层[内夫等人(1983年)。《发育生物学》97卷,第103 - 112页;布莱克和格哈特(1985年)。《发育生物学》108卷,第310 - 324页]。我们通过细胞质移植证明,背侧活性特异性地定位于倒置卵的未着色皮层细胞质中,该细胞质被分离到未来的外胚层谱系中。这一结果表明,背侧决定因子与未着色皮层相关,并且不会因倒置而移位。此外,我们发现两个植物性定位的转录本,Xcat2和Vg1 mRNA,存在于倒置卵的反转动物极中,表明它们与未着色皮层相关。为了将背侧决定因子活性与已知的背侧化分子进行比较,我们检查了反转胚胎中Xnr3和Siamois的表达模式,因为这两个基因是由Wnt信号通路激活剂(Xwnt - 8、β - 连环蛋白等)激活的,而不是由其他背侧化分子(noggin、BVg1等)激活的。接受卵未着色皮层的反转胚胎的动物帽表达Xnr3和Siamois。然而,Mix.1,一种在正常胚胎中响应中胚层诱导剂在内胚层和中胚层中表达的标记物,在反转胚胎的动物帽中未被检测到。此外,我们发现β - 连环蛋白在反转胚胎未着色动物极细胞的细胞核中积累。这些结果表明,母体背侧决定因子的行为与Wnt信号通路激活剂比与noggin或BVg1更相似。