Characterization of a vasa homolog in the brown-marbled grouper (Epinephelus fuscoguttatus) and its expression in gonad and germ cells during larval development

The vasa gene is specifically expressed in the germ cell lineage, and its expression has been used to study germline development in many organisms, including fishes. In this study, we cloned and characterized vasa as Efu-vasa in the brown-marbled grouper (Epinephelus fuscoguttatus). Efu-vasa contained predicted regions that shared consensus motifs with the vasa family in teleosts, including arginine- and glycine-rich repeats, ATPase motifs, and a DEAD box. Phylogenetic-tree construction using various DEAD-box proteins confirmed that Efu-vasa was clustered in the vasa family. Efu-vasa mRNA was detectable only in gonads, by reverse transcription polymerase chain reaction. Primordial germ cells (PGCs) during early gonad development in larvae were characterized by histological examination and in situ hybridization using an Efu-vasa antisense probe. Migrating PGCs were found in larvae at 9-21 days post-hatching, and rapid proliferation of PGCs was initiated in 36 days post-hatching. These findings provide a valuable basis for optimizing the developmental stages for germ cell transplantation in order to produce surrogate broodstock, which may help in the production of larvae of large and endangered grouper species.