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Updated: Sep 5, 2025
In Vivo Functional Brain Imaging Approach Based on Bioluminescent Calcium Indicator GFP-aequorin
Published on: January 8, 2016
Optimized Aequorin Reconstitution Protocol to Visualize Calcium Ion Transients in the Heart of Transgenic Zebrafish
Manuel Vicente1, Jussep Salgado-Almario1, Antonio Martínez-Sielva1
1Physiology and Cell Dynamics Group, Centro Regional de Investigaciones Biomédicas (CRIB) and Facultad de Medicina de Albacete, Universidad de Castilla-La Mancha, Albacete, Spain.
Abstract:
We introduce how to image calcium ion levels in the heart of zebrafish embryos and larvae up to 5 days post-fertilization with the photoprotein green fluorescent protein (GFP)-aequorin (GA) in the transgenic line Tg(myl7:GA). Incubation of the embryos with CTZ to obtain the functional photoprotein yields few emission counts, suggesting that, when the heart is beating, the rate of aequorin consumption is faster than that of the reconstitution with CTZ. In this chapter, we present an improved aequorin reconstitution protocol. We further describe the experimental procedure as well as the bioluminescence data analysis and processing.
