Université Pierre et Marie Curie and Centre National de la Recherche Scientifique, BioMarCell, UMR7009 BioDev, Observatoire Océanologique, Villefranche-sur-mer, 06234, France.
Dev Biol. 2009 Dec 1;336(1):96-111. doi: 10.1016/j.ydbio.2009.09.001. Epub 2009 Sep 6.
Ascidian postplasmic/PEM RNAs constitute a large class of cortical maternal RNAs which include developmental determinants (macho-1 and pem-1). We have analyzed the localization, cortical anchorage and cell type segregation of postplasmic/PEM RNAs in Ciona intestinalis and Phallusia mammillata using very high-resolution fluorescent in situ hybridization. We also compared RNAs extracted from whole oocytes and from isolated cortices using microarrays and localized RNAs possessing clusters of xCACx motifs in their 3'UTRs. Based on these combined approaches we conclude that: (1) the vast majority of the 39 postplasmic/PEM RNAs (including vasa) are localized in the egg cortex. (2) Many postplasmic/PEM RNAs 3'UTR are enriched in xCACx motifs, allowing us to identify 2 novel postplasmic/PEM RNAs (PSD and MnK). (3) Postplasmic/PEM RNAs anchored to cortical Endoplasmic Reticulum (cER) and those associated with granules have different cell destinations. We propose that there are 2 distinct categories of postplasmic/PEM RNAs on the basis of their cortical anchorages and cell destinations: (1) macho-1-like postplasmic/PEM RNAs anchored to cER which segregate into somatic B8.11 cells. (2) vasa-like postplasmic/PEM RNAs associated with granules which in addition to B8.11 cells segregate into B8.12 germ cells.
海鞘细胞质/PEM RNA 构成了一大类皮质母体 RNA,其中包括发育决定因素(macho-1 和 pem-1)。我们使用超高分辨率荧光原位杂交技术分析了海鞘和 Phallusia mammillata 中细胞质/PEM RNA 的定位、皮质锚定和细胞类型分离。我们还使用微阵列比较了从整个卵母细胞和分离的皮质中提取的 RNA,并定位了在其 3'UTR 中具有 xCACx 基序簇的 RNA。基于这些综合方法,我们得出结论:(1) 39 种细胞质/PEM RNA(包括 vasa)中的绝大多数定位于卵母细胞皮质。(2) 许多细胞质/PEM RNA 的 3'UTR 富含 xCACx 基序,使我们能够识别 2 种新的细胞质/PEM RNA(PSD 和 MnK)。(3) 锚定在皮质内质网 (cER) 上的细胞质/PEM RNA 和与颗粒相关的 RNA 具有不同的细胞归宿。我们提出,根据它们的皮质锚定和细胞归宿,细胞质/PEM RNA 有 2 种不同的类别:(1) macho-1 样细胞质/PEM RNA 锚定在 cER 上,分离为体 B8.11 细胞。(2) 与颗粒相关的 vasa 样细胞质/PEM RNA 除了 B8.11 细胞外,还分离为 B8.12 生殖细胞。