Localization of germline marker vasa homolog RNA to a single blastomere at early cleavage stages in the oriental river prawn Macrobrachium nipponense: Evidence for germ cell specification by preformation

Abstract

Germ cells are specified by the inheritance of maternal germline determinants (preformation mode) or inductive signals from somatic cells (epigenesis mode) during embryogenesis. However, the germline specification in decapod crustaceans is unclear so far. Using vasa homolog (MnVasa) as a germ cell marker, here we probed the early events of germline specification in the oriental river prawn Macrobrachium nipponense. Quantitative RT-PCR analysis of unfertilized eggs and embryos demonstrated that the prawn MnVasa mRNA is a maternal factor. Whole-mount in situ hybridization further indicated that MnVasa transcripts are maternally supplied to only one blastomere at the very early cleavage stages. As cleavage proceeds, the MnVasa-positive blastomere undergoes proliferation and increases in number. During gastrulation, the MnVasa-positive cells are found to be around a blastopore and could migrate into an embryo through the blastopore. At the zoea stage, clusters of the MnVasa-positive cells distribute not only in the gonad rudiment in the cephalothorax but also at an extragonadic site, dorsal to the posterior hindgut in the abdomen, suggesting that MnVasa-positive cells could migrate anteriorly to the genital rudiment through the hindgut. Based on the dynamic localization and number of MnVasa-positive cells during embryogenesis, we concluded that the MnVasa-positive cells are primordial germ cells (PGC) or founder cells of PGC that are separated from soma at the early cleavage stage. MnVasa mRNA might have a key function in the specification of the prawn germline cells as a maternal determinant. These results provide the first evidence that the germline specification in decapod crustaceans follows a preformation mode.