Compensatory lymphangiogenesis is required for edema resolution in zebrafish

Edema, characterized by the accumulation of interstitial fluid, poses significant challenges in various pathological conditions. Lymphangiogenesis is critical in edema clearance, and delayed or inadequate lymphatic responses significantly hinder healing processes. However, real-time observation of dynamic changes in lymphangiogenesis during tissue repair in animal models has been challenging, leaving the mechanisms behind compensatory lymphatic activation for edema clearance largely unexplored. To address this gap, we subjected zebrafish larvae to osmotic stress using hypertonic (375 mOsm/L) and isotonic (37.5 mOsm/L) solutions to induce osmotic imbalance and subsequent edema formation. Intravital imaging of vascular transgenic larvae revealed significant lymphatic vessel remodeling during tissue edema. The observed increase in lymphatic endothelial progenitor cells, alongside the sustained expansion and remodeling of primary lymphatics, indicates active lymphangiogenesis during the recovery phase. We developed a novel method employing translating ribosome affinity purification to analyze the translatome of lymphatic and venous endothelial cells in vivo, which uncovered the upregulation of key pro-lymphangiogenic genes, particularly vegfr2 and vegfr3, during tissue recovery. Inhibition of compensatory lymphangiogenesis impaired edema fluid clearance and tissue recovery. Our findings establish a new model for in vivo live imaging of compensatory lymphangiogenesis and provide a novel approach in investigating lymphatic activation during edema resolution.