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© Research
Publication : Chemical Science

Ultraspecific live imaging of the dynamics of zebrafish neutrophil granules by a histopermeable fluorogenic benzochalcone probe

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Chemical Science - 14 Feb 2019

Colucci-Guyon E, Batista AS, Oliveira SDS, Blaud M, Bellettini IC, Marteyn BS, Leblanc K, Herbomel P, Duval R

Link to Pubmed [PMID] – 30996961

Chem Sci 2019 Mar;10(12):3654-3670

Neutrophil granules (NGs) are key components of the innate immune response and mark the development of neutrophilic granulocytes in mammals. However, there has been no specific fluorescent vital stain up to now to monitor their dynamics within a whole live organism. We rationally designed a benzochalcone fluorescent probe () featuring high tissue permeability and optimal photophysics such as elevated quantum yield, pronounced solvatochromism and target-induced fluorogenesis. Phenotypic screening identified as the first cell- and organelle-specific small-molecule fluorescent tracer of NGs in live zebrafish larvae, with no labeling of other cell types or organelles. staining was independent of the state of neutrophil activation, labeling NGs of both resting and phagocytically active neutrophils with equal specificity. By high-resolution live imaging, we documented the dynamics of -stained NGs during phagocytosis. Upon zymosan injection, labeled NGs were rapidly recruited to the forming phagosomes. Despite being a reversible ligand, could not be displaced by high concentrations of pharmacologically relevant competing chalcones, indicating that this specific labeling was the result of the ‘s precise physicochemical signature rather than a general feature of chalcones. However, one of the competitors was discovered as a promising interstitial fluorescent tracer illuminating zebrafish histology, similarly to BODIPY-ceramide. As a yellow-emitting histopermeable vital stain, functionally and spectrally complements most genetically incorporated fluorescent tags commonly used in live zebrafish biology, holding promise for the study of neutrophil-dependent responses relevant to human physiopathology such as developmental defects, inflammation and infection. Furthermore, intensely labeled isolated live human neutrophils at the level of granulated subcellular structures consistent with human NGs, suggesting that the labeling of NGs by is not restricted to the zebrafish model but also relevant to mammalian systems.

https://www.ncbi.nlm.nih.gov/pubmed/30996961