Embryonic expression of p68, a DEAD-box RNA helicase, in the oligochaete annelid Tubifex tubifex.

We have cloned and characterized the expression of a p68 homologue (designated Ttu-p68) from the oligochaete annelid Tubifex tubifex. Ttu-p68 mRNA is distributed broadly throughout the early stages. Ttu-p68 is expressed in all of the early blastomeres, in which Ttu-p68 RNA associates with pole plasms. Ttu-p68 transcripts are concentrated to 4d cell but not to 2d cell. During gastrulation, expression of Ttu-p68 is restricted to elongating germ bands (GBs) and an anteriormost crescent of micromere descendants on both sides of the embryo. During body elongation that follows gastrulation, expression of Ttu-p68 is further restricted to the stomodaeum (derived from the micromere crescent), ventral ganglia, lateral dots (corresponding to dorsal and ventral setal sacs), ventral large cells (that resemble presumptive primordial germ cells) in segments VIII-XII, and a bilateral pair of cell clusters at the caudal end. At the end of embryogenesis, Ttu-p68 expression persists exclusively in the tail and the lining epithelium of the pharynx.

Transient occurrence of vasa-expressing cells in nongenital segments during embryonic development in the oligochaete annelid Tubifex tubifex.

The primordial germ cells (PGCs) in the oligochaete annelid Tubifex tubifex are mesodermal in origin and are located in the two midbody segments X and XI in which the testis and the ovary are formed, respectively. To identify a molecular marker for the Tubifex PGCs, we isolated the Tubifex homologue (Ttu-vas) of the Drosophila vasa gene. Using whole-mount in situ hybridization, we examined the spatial expression patterns of Ttu-vas from one-cell stage through juvenile stage. Ttu-vas messenger ribonucleic acid (RNA) is present as a maternal transcript distributed broadly throughout the early stages. Ttu-vas is expressed in all of the early cleavage blastomeres, in which Ttu-vas RNA associates with mitotic spindles and pole plasms. Expression of Ttu-vas gradually becomes restricted, first to teloblasts, then to their blast cell progeny comprising the germ bands (GBs), and finally to a set of large ventral cells (termed VE cells) in a variable set of midbody segments including the genital segments (X and XI). At the end of embryogenesis, VE cells are confined to genital segments where they are presumably germline precursors in the juvenile. Staining with a cross-reacting anti-Vasa antibody suggested that VE cells express Ttu-vas protein to the same extent irrespective of their positions along the anteroposterior axis. A set of cell ablation experiments suggested that VE cells are derived from the mesodermal teloblast lineage and that the emergence of VE cells takes place independently of the presence of the ectodermal GBs that normally overlay the mesoderm. These results suggest that T. tubifex generates supernumerary presumptive PGCs during embryogenesis whose number is variable among embryos.