Abstract
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.
Highlights
Adult zebrafish, in contrast to mammals, are able to regenerate their hearts in response to injury or experimental amputation
Enrichment of Zebrafish Heart extracellular matrix (ECM) Proteins—To study the ECM protein composition of zebrafish ventricles, we developed a decellularization protocol consisting on detergent treatment with 0,5% sodium dodecyl sulfate (SDS), followed by 1% Triton-X 100 to remove sodium-dodecyl disulfate (SDS) prior to processing for analysis (Fig. 1A)
To analyze whether the decellularization process altered the relative abundance of ECM proteins, we characterized the proteome of zebrafish hearts before, after SDS treatment, and at the end of decellularization by liquid chromatographymass spectrometry (LC-MS)
Summary
In contrast to mammals, are able to regenerate their hearts in response to injury or experimental amputation. Our cumulative results identify changes in the protein composition and mechanical properties of the zebrafish heart ECM during regeneration. Gene expression analysis performed in regenerating hearts identified transcripts encoding ECM-related proteins among the most differentially expressed during this process (44 – 46).
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