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

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Abstract

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.

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Acknowledgments

We are grateful to Drs. C. G Extavour and M. Akam, University of Cambridge, for the Vasa antibody. We also thank members of the Shimizu laboratory for advice and help in collecting embryos. This study was supported in part by a Grant-in-Aid from the Ministry of Education, Science, Sports and Culture, Japan (13680799) to T.S.

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  1. Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo, 060-0810, Japan

    Atsuko Oyama & Takashi Shimizu

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  1. Atsuko Oyama
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  2. Takashi Shimizu
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Correspondence to Takashi Shimizu.

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Communicated by: D.A. Weisblat

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Fig. S1

Characterization of Ttu-vas, a vasa homologue from T. tubifex. a Alignment of Ttu-Vas with known Vasa-class proteins. The eight conserved motifs of the DEAD-box protein family are boxed in black. The EARKF and WD motifs are boxed in red. Numbers in parentheses indicate the percentage amino acid identity with the overall sequences of Ttu-Vas. b Molecular phylogenetic relationship of Ttu-Vas to other DEAD-box proteins. The phylogenetic tree was generated by the neighbor joining method using PAUP*4.0b10. Dme-Abs was used as an outgroup. Numbers are bootstrap values (as percentages of 1000 replications). Lengths of branches are drawn to the scale indicated (Bmo Bombyx mori, Cgi Crassostrea gigas, Csa Ciona savignyi, Dma Daphnia magna Dme Drosophila melanogaster Dre Danio rerio, Gga Gallus gallus, Hma Hydra magnipapillata, Mmu Mus musculus, Ola Oryzias latipes, Pdu Platynereis dumerilii, Ttu Tubifex tubifex) (GIF 118 445 KB)

High-resolution image (TIFF 7.43 MB)

Fig. S2

Frequency of total number of ventral Ttu-vas-expressing cells present on the left (a) and right (b) side of individual embryos. Abscissa: developmental stages (16a to 18) and juvenile stages (Juv-D1 to D3) (GIF 28 879 kb)

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Oyama, A., Shimizu, T. Transient occurrence of vasa-expressing cells in nongenital segments during embryonic development in the oligochaete annelid Tubifex tubifex . Dev Genes Evol 217, 675–690 (2007). https://doi.org/10.1007/s00427-007-0180-1

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