Browsing by Author "Cross, I."
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- Solea senegalensis vasa transcripts: molecular characterisation, tissue distribution and developmental expression profilesPublication . Pacchiarini, T.; Cross, I.; Leite, Ricardo; Gavaia, Paulo J.; Ortiz-Delgado, J. B.; Pousão-Ferreira, P.; Rebordinos, L.; Sarasquete, C.; Cabrita, ElsaThe Vasa protein is an RNA helicase belonging the DEAD (Asp-Glu-Ala-Asp)-box family. The crucial role played by the vasa gene in the germ-cell lineage of both vertebrates and invertebrates has made this gene a useful molecular marker for germinal cells and a useful tool in surrogate broodstock production using primordial germ cell transplantation. With the aim of establishing a novel approach to improving Solea senegalensis broodstock management, the vasa gene in this species was characterised. Four S. senegalensis vasa transcripts were isolated: Ssvasa1, Ssvasa2, Ssvasa3 and Ssvasa4. Their phylogenetic relationship with other vasa homologues was determined confirming the high degree of conservation of this helicase throughout evolution. Our qPCR results showed that S. senegalensis vasa transcripts are prevalently expressed in gonads, with ovary-specific expression for Ssvasa3 and Ssvasa4. During embryonic and larval development, a switch between the longest and the shortest transcripts was observed. While Ssvasa1 and Ssvasa2 were maternally supplied, Ssvasa3 and Ssvasa4 depended on the de novo expression program of the growing juveniles, suggesting that vasa mRNA could be involved in Senegalese sole gonad differentiation. In situ hybridisation and immunohistochemical analysis performed in 150-days after hatching (DAH) larvae showed vasa product expression in the germinal region of early gonads. In our work we demonstrated the usefulness of Ssvasa mRNAs as molecular markers for primordial germ cells and germinal cells during embryonic development, larval ontogenesis and gonad differentiation. Furthermore, our results confirmed the potential of vasa to help investigate germinal cell biotechnology for Senegalese sole reproduction.
- Two isoforms of vasa maternal factor in Senegalese sole: Biotechnological applicationsPublication . Pacchiarini, T.; Cabrita, Elsa; Cross, I.; Ortiz-Delgado, J. B.; Leite, Ricardo; Gavaia, Paulo J.; Herráez, M. P.; Rebordinos, L.; Sarasquete, C.Primordial Germ Cells (PGCs) identification and manipulation present considerable potential for hatchery practice and surrogate broodstocks. To carry out the PGCs characterization a specific molecular marker is required. The vasa gene is a good candidate to identify PGCs and others germinal cells (Nagasawa et al., 2009). The aim of this study was the cloning of the Solea senegalensis vasa cDNA and its expression pattern during early development and adulthood.