A genetically encoded sensor for visualizing leukotriene B4 gradients in vivo.

Abstract

Leukotriene B₄ (LTB₄) is a potent lipid chemoattractant driving inflammatory responses during host defense, allergy, autoimmune and metabolic diseases. Gradients of LTB₄ orchestrate leukocyte recruitment and swarming to sites of tissue damage and infection. How LTB₄ gradients form and spread in live tissues to regulate these processes remains largely elusive due to the lack of suitable tools for monitoring LTB₄ levels in vivo. Here, we develop GEM-LTB₄, a genetically encoded green fluorescent LTB₄ biosensor based on the human G-protein-coupled receptor BLT1. GEM-LTB₄ shows high sensitivity, specificity and a robust fluorescence increase in response to LTB₄ without affecting downstream signaling pathways. We use GEM-LTB₄ to measure ex vivo LTB₄ production of murine neutrophils. Transgenic expression of GEM-LTB₄ in zebrafish allows the real-time visualization of both exogenously applied and endogenously produced LTB₄ gradients. GEM-LTB₄ thus serves as a broadly applicable tool for analyzing LTB₄ dynamics in various experimental systems and model organisms.