Copper-induced stress and recovery impacts on organismal phenotypes and the underlying proteomic signatures in Botryllus schlosseri

This study establishes the copper tolerance range of the colonial marine tunicate Botryllus schlosseri. Furthermore, quantitative organismal phenotyping and quantitative proteomics were combined to characterize the B. schlosseri response to, and recovery from, acute copper exposure stress. Changes in the area of B. schlosseri colony systems and pigmentation provided sensitive, dose-dependent markers of exposure to, and recovery from, copper stress. Comprehensive quantitative proteomics using consistent data-independent acquisition (DIA) assay libraries revealed activation of detoxification, oxidative stress, and immune pathways during exposure to copper stress. In addition, quantitative proteomics uncovered enrichment of tissue remodeling and proliferative signaling pathways during recovery from copper stress. We identified 35 proteins whose expression closely mirrored phenotypic changes observed at the colonial system level. This specific proteome signature represents a comprehensive molecular underpinning of the organismal response of B. schlosseri to copper stress. In conclusion, this study establishes copper tolerance ranges of the invasive colonial tunicate B. schlosseri and explains the molecular underpinnings of stress-induced organismal phenotypes by identifying corresponding proteome signatures and their associated functional enrichments. Moreover, identification of copper concentrations that are stressful and highly disruptive on the molecular phenotype, yet readily recoverable from, lays a critical foundation for future studies directed at stress-induced adaptation and evolutionary trajectories of marine invertebrates in changing and novel environments.