Proteomics Analysis of Extracellular Matrix Remodeling During Zebrafish Heart Regeneration*

Zebrafish can regenerate their hearts. The role of the extracellular matrix in this process is largely unknown. We have analyzed the proteome in control hearts and at different times of regeneration. Decellularization of samples allowed for enrichment of extracellular matrix proteins, increasing their detection. The results reported dynamic changes in specific proteins associated with specific stages of the regenerative process. Biomechanical analysis by atomic force microscopy revealed concomitant changes in matrix stiffness during this process. Graphical Abstract Highlights We have developed a decellularization protocol for ECM protein enrichment. We have characterized the proteome of adult zebrafish heart ECM. We describe dynamic changes in heart ECM proteome during regeneration. We describe changes in heart ECM stiffness during regeneration. Adult zebrafish, in contrast to mammals, are able to regenerate their hearts in response to injury or experimental amputation. Our understanding of the cellular and molecular bases that underlie this process, although fragmentary, has increased significantly over the last years. However, the role of the extracellular matrix (ECM) during zebrafish heart regeneration has been comparatively rarely explored. Here, we set out to characterize the ECM protein composition in adult zebrafish hearts, and whether it changed during the regenerative response. For this purpose, we first established a decellularization protocol of adult zebrafish ventricles that significantly enriched the yield of ECM proteins. We then performed proteomic analyses of decellularized control hearts and at different times of regeneration. Our results show a dynamic change in ECM protein composition, most evident at the earliest (7 days postamputation) time point analyzed. Regeneration associated with sharp increases in specific ECM proteins, and with an overall decrease in collagens and cytoskeletal proteins. We finally tested by atomic force microscopy that the changes in ECM composition translated to decreased ECM stiffness. Our cumulative results identify changes in the protein composition and mechanical properties of the zebrafish heart ECM during regeneration.

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