Conclusions
This study presents a novel experimental design combined with an
innovative research tool for systems-level analyses of molecular
phenotypes (DIA-LCMS2). Specific organismal phenotypes (blood
osmolality, body condition, feeding rate) were found to correlate well
with extreme salinity stress in O. mossambicus . The
rate-independent salinity maximum and critical threshold salinity were
determined using a comprehensive set of acclimations and a generalized
protocol that is reproducible for other species. The DIA-LCMS2 approach
was shown to reveal comprehensive molecular phenotypes in ecologically
relevant samples. The data generated in this study were used to produce
protein networks which elucidate the mechanisms of ionic and
mitochondrial protein regulation in response to hypersalinity in general
and with respect to the critical threshold. These networks reveal novel
information regarding the modulation of energy metabolism, cell
adhesion, and cell structure, which includes functional annotation of
previously uncharacterized proteins.