ELUCIDATING THE ROLE OF MIR-10A/B IN THE WOUND HEALING PROCESS IN LARVAL ZEBRAFISH

Abstract

This study explores the regulatory role of microRNA-10 (miR-10) in larval zebrafish caudal fin regeneration, a model with broad implications for the future of regenerative medicine. Building on prior research from the lab, which indicated that miR-10 knockdown impairs regeneration, we verified this finding through rescue experiments, assessed its influence on healing projections, and examined its role in the innate immune response during the wound healing process. To address these questions, we employed a multi-faceted approach, including timed matings, microinjections, immunohistochemistry, fixed sample microscopy, and live imaging. Our results show that the experimental introduction of exogenous miR-10 can rescue the regenerative defect caused by morpholino-mediated knockdown. In addition, miR-10 participates in cellular proliferation without altering apoptosis during the healing process, further eliminating off-target concerns for the morpholino. miR-10 also influences the formation of healing projections, as confocal imaging revealed a decrease in vimentin expression at the wound site 20 hours post-injury in miR-10-deficient larvae. This suggests a potential link between miR-10 and vimentin expression, a known regulator of healing projections. Additionally, knockdown of miR-10 reduced both overall neutrophil and macrophage development, as evidenced by total cell counts at 2 days post-fertilization, suggesting a role for miR-10a/b in proper hematopoiesis in developing zebrafish. Not surprisingly, a marked decrease in leukocyte recruitment to the wound site was also observed. Together, these findings highlight miR-10 as a key regulator of the healing process, particularly through its roles in modulating immune cell dynamics and promoting healing projections