Proteomic analysis through larval development of Solea senegalensis flatfish

Abstract

The post-embryonic development of the Senegalese sole, Solea senegalensis, a flatfish of growing interest in fisheries and aquaculture, is associated with drastic morpho-physiological changes during metamorphosis. Although in the last two decades knowledge on sole culture has notably increased, especially in Southern Europe, its progress was restricted due to lack of methods to control reproduction, improve larval quality and increase juvenile disease resistance. A limited knowledge of the physiological, molecular and genetic mechanisms involved is at the base of such limitation. A proteomic study was carried out to explore the molecular events that occur during S. senegalensis ontogenesis. Protein expression changes were monitored in larvae from 5 to 21 dph by combining 2DE and protein identification with de novo MS/MS sequencing. An average of 6177 ± 282 spots was resolved in 2DE gels. Hierarchical cluster analysis of the 705 selected spots grouped them in eight patterns. Thirty-four proteins were identified and assigned biological functions including structure, metabolism highlighting energy metabolism, transport, protein folding, stress response, chromatin organization and regulation of gene expression. These changes provide a sequential description of the molecular events associated with the biochemical and biological transformations that occur during sole larval development.

Sequential protein extraction as an efficient method for improved proteome coverage in larvae of Atlantic salmon ( Salmo salar )

“…The production of high‐quality salmon larvae is a key target of commercial aquaculture because the larval phase is a critical period that influences further predispositions during the remaining life cycle . Proteomics can be applied to enhance our understanding of physiological responses to environmental and dietary factors that determine larval quality . However, as with any tissue or organism comprising a highly complex mixture of cellular proteins and tissues dominated by abundant structural components, depth of proteome coverage is hampered by the very large dynamic range of protein abundance.…”

“…that determine larval quality [3]. However, as with any tissue or organism comprising a highly complex mixture of cellular proteins and tissues dominated by abundant structural components, depth of proteome coverage is hampered by the very large dynamic range of protein abundance.…”

“…While fractionation of proteins based on differential solubility has been used effectively to reduce sample complexity prior to 2DE analysis (for example in ), it is not commonly used in shotgun proteomics studies. Proteomics‐based analysis of commercial fish larvae has generally utilized 2DE after direct protein extraction in denaturing buffer , but this approach falls short in terms of reproducibility and detection of lower‐abundance proteins . Here, we used label‐free quantitative proteomics to determine the effectiveness of sequential extraction (SE) based on differential protein solubility as an effective strategy to increase proteome coverage of whole fish salmon larvae.…”