Molecular Characterization of Embryos with Different Buoyancy Levels in the Yellowtail Kingfish (Seriola lalandi)

SIMPLE SUMMARY: Low survival of embryos in Seriola lalandi captivity farming has been attributed to low buoyancy. This process is the result of oocyte hydration, which depends on the osmotic force exerted by free amino acids (FAA) generate of cathepsin-mediated yolk proteins proteolysis. In order to understand the molecular bases of buoyancy acquisition process and its loosing throughout the development of S. lalandi, the aim of this study was to compare gene expression and activity of cathepsins, as well as the FAA content between floating and low-floating embryos. Eggs, morula, blastula, gastrula and 24 h embryos were the stages collected in this study. These assessments were supplemented with morphometric and functional characterization of the embryos, where no differences in embryo and oil drop diameter, and constitutive gene expression were detected between floating and low-floating embryos. Cathepsin B did not show differences in expression or activity related to buoyancy condition. Both expression and activity of cathepsin D were higher in some low-floating developmental stages. By contrast, cathepsin L showed higher expression and activity in some floating early embryos. Higher FAA content was observed in floating embryos at least until gastrula stage in comparison to low-floating ones. In summary, expression and activity of cathepsins and FAA content, revealed specific pattern throughout development or buoyancy conditions of the embryos. This study identifies molecular differences between floating and low floating embryos at specific developmental stages where cathepsins and FAA are promising markers to evaluate the embryo quality in the farming of this species. ABSTRACT: The buoyancy of eggs and embryos is associated with successful development in pelagic fish. Buoyancy is the result of oocyte hydration, which depends on the osmotic force exerted by free amino acids (FAA) generated by yolk proteolysis, and cathepsins are the main enzymes involved in this process. Seriola lalandi is a pelagic fish whose farming has been hampered by development failure that have been partially attributed to decreased buoyancy of embryos. Therefore, the aim of this study was to compare the mRNA expression and activity of cathepsins B, D, and L, as well as the FAA content in floating and low-floating embryos at different developmental stages. The chosen stages were eggs, morula, blastula, gastrula and 24 h embryos. Complementary assessments showed that there were no differences attributed to buoyancy status in embryo and oil droplet diameters, as well as the transcriptional status at any developmental stage. Cathepsin B did not show differences in mRNA expression or activity related to buoyancy at any stage. Cathepsin D displayed higher transcript and activity levels only in low-floating eggs compared with those floating. Cathepsin L showed higher expression in floating eggs and 24 h embryos compared with that of low-floating, but the activity of this enzyme was higher in floating eggs and morula. Total FAA content constantly decreased throughout development in floating embryos, but it was always higher than low-floating embryos until gastrula stage. In 24 h embryos floating and low-floating embryos share similar quantities of FAA. In summary, differences in the expression and activity of cathepsins between floating and low-floating embryos could be revealed at specific embryonic stages, suggesting different functions of these enzymes throughout development. Besides 24 h embryos, FAA content seems to be a decisive factor for buoyancy of embryos during early development of S. lalandi. Overall, considering the main role of cathepsins and FAA in buoyancy acquisition process and therefore in both embryo quality and viability, our study identifies good marker candidates to evaluate embryo quality in the farming of this species.