Authors:

Erbol Nishanov

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About Paper:

Growing evidence suggests that bioelectricity modulates embryonic development, tissue regeneration, and congenital diseases. Recent invention of genetically encoded voltage indicators (GEVIs) surmounted the challenge of non-invasive in vivo assessment of cellular electric potential. Voltron, a novel genetically encoded voltage indicator, implements the HaloTag protein complex to bind to the bright and photostable synthetic fluorophore ligand dyes. The functional versatility of these dyes makes Voltron a superior indicator, allowing for brighter, high-resolution, prolonged imaging. The goal of our research is to investigate regeneration biology of the zebrafish caudal fin. A previously established transgenic Voltron zebrafish line with UAS promoter was bred with Tg(and1: Gal4FF) to image the bioelectric activities of zebrafish larva caudal fins. The embryos at the early development stage were incubated in the working solution of the selected ligand dye and then paralyzed and mounted in low-melting agarose for imaging. Movies of various duration were obtained to document the change of the fluorescence intensity in the fish's caudal fin. The fluorescence data were quantified using Image J and analyzed via MATLAB and Minitab. Resulting data will be used to assess the behavior of the electric potential in the embryos. Upon successful application of the Voltron GEVI, the imaging technique can be used to reveal the role of bioelectricity in the fin development and regeneration and its relationship with calcium signaling.

Source:

Purdue University / 2023

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Co-authors:

Erbol Nishanov