The germline of multicellular animals is segregated from somatic tissues, which is an essential developmental process for the next generation. Although certain ecdysozoans and chordates segregate their germline during embryogenesis, animals from other taxa segregate their germline after embryogenesis from multipotent progenitor cells. An overlapping set of genes, including vasa, nanos and piwi, operate in both multipotent precursors and in the germline. As we propose here, this conservation implies the existence of an underlying germline multipotency program in these cell types that has a previously underappreciated and conserved function in maintaining multipotency.

Echinoderms (sea urchins, sea stars, brittle stars, sea lilies and sea cucumbers) are a group of diverse organisms, second in number within deuterostome species to only the chordates. Echinoderms serve as excellent model systems for developmental biology due to their diverse developmental mechanisms, tractable laboratory use, and close phylogenetic distance to chordates. In addition, echinoderms are very well represented in the fossil record, including some larval features, making echinoderms a valuable system for studying evolutionary development. The internal relationships of Echinodermata have not been consistently supported across phylogenetic analyses, however, and this has hindered the study of other aspects of their biology. In order to test echinoderm phylogenetic relationships, we sequenced 23 de novo transcriptomes from all five clades of echinoderms. Using multiple phylogenetic methods at a variety of sampling depths we have constructed a well-supported phylogenetic tree of Echinodermata, including support for the sister groups of Asterozoa (sea stars and brittle stars) and Echinozoa (sea urchins and sea cucumbers). These results will help inform developmental and evolutionary studies specifically in echinoderms and deuterostomes in general.

Many indirect developing animals create specialized multipotent cells in early development to construct the adult body and perhaps to hold the fate of the primordial germ cells. In sea urchin embryos, small micromeres formed at the fifth division appear to be such multipotent cells: they are relatively quiescent in embryos, but contribute significantly to the coelomic sacs of the larvae, from which the major tissues of the adult rudiment are derived. These cells appear to be regulated by a conserved gene set that includes the classic germline lineage genes vasa, nanos and piwi. In vivo lineage mapping of the cells awaits genetic manipulation of the lineage, but previous research has demonstrated that the germline is not specified at the fourth division because animals are fertile even when micromeres, the parent blastomeres of small micromeres, are deleted. Here, we have deleted small micromeres at the fifth division and have raised the resultant larvae to maturity. These embryos developed normally and did not overexpress Vasa, as did embryos from a micromere deletion, implying the compensatory gene regulatory network was not activated in small micromere-deleted embryos. Adults from control and micromere-deleted embryos developed gonads and visible gametes, whereas small micromere-deleted animals formed small gonads that lacked gametes. Quantitative PCR results indicate that small micromere-deleted animals produce background levels of germ cell products, but not specifically eggs or sperm. These results suggest that germline specification depends on the small micromeres, either directly as lineage products, or indirectly by signaling mechanisms emanating from the small micromeres or their descendants.