Uncovering injury-specific proteomic signatures and regenerative decline risks in single and repetitive appendage trauma
June 23
Repetitive soft-tissue trauma is increasingly recognised as a driver of chronic, non-healing lesions and late-onset degenerative pathologies, yet the molecular cascades triggered by recurrent injury remain poorly defined. Here, we delineate proteomic and functional consequences of single versus repetitive mild caudal-fin resection in the adult zebrafish, concentrating on acute (48 h) and subacute (1 week) phases within the wound epidermis and blastema.
Using unbiased shotgun proteomics integrated with weighted gene co-expression network analysis and machine-learning classifiers, we mapped the injury-dependent protein landscapes of the regenerating stump. Morphometric and contractile performance assays were conducted at 2, 7 and 30 days post-injury to couple molecular alterations to regenerative outcomes.
Spatio-temporal analyses revealed distinct, injury-number–specific proteomic programmes. A single resection activated pro-regenerative modules encompassing proliferative, patterning and extracellular matrix remodelling factors, whereas repetitive trauma amplified oxidative stress, mitochondrial dysfunction and gap-junction disruption within the blastema. Notably, Apoa1, ApoE, Cox6a1 and Snca emerged as hub proteins whose dysregulation correlated with delayed blastemal consolidation and scar-prone matrix deposition, linking recurrent injury to pathways classically associated with age-related tissue degeneration.
Functional testing corroborated the molecular data: repetitive injury provoked significant deficits in fin regrowth rate and swimming thrust at days 2 and 7. Although contractile performance normalised by day 30, length restitution and radial-ray patterning remained incomplete, indicating persistent morphogenetic impairment.
Collectively, these results furnish a high-resolution portrait of the proteomic networks governing effective versus failed appendage regeneration and nominate mitochondrial resilience and intercellular coupling as actionable targets for preventing long-term regenerative decline after recurrent tissue trauma.