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In vivo imaging of calcium dynamics in zebrafish hepatocytes

View ORCID ProfileMacarena Pozo-Morales, View ORCID ProfileInés Garteizgogeascoa, Camille Perazzolo, View ORCID ProfileSumeet Pal Singh
doi: https://doi.org/10.1101/2022.01.11.475536
Macarena Pozo-Morales
1IRIBHM, Université Libre de Bruxelles (ULB), Route de Lennik 808, 1070 Brussels, Belgium
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Inés Garteizgogeascoa
1IRIBHM, Université Libre de Bruxelles (ULB), Route de Lennik 808, 1070 Brussels, Belgium
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Camille Perazzolo
1IRIBHM, Université Libre de Bruxelles (ULB), Route de Lennik 808, 1070 Brussels, Belgium
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Sumeet Pal Singh
1IRIBHM, Université Libre de Bruxelles (ULB), Route de Lennik 808, 1070 Brussels, Belgium
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  • For correspondence: sumeet.pal.singh@ulb.be
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ABSTRACT

Hepatocytes were the first cell-type for which oscillations of cytoplasmic calcium levels in response to hormones were described. Since then, investigation of calcium dynamics in liver explants and culture has greatly increased our understanding of calcium signaling. A bottleneck, however, exists in observing calcium dynamics in a non-invasive manner due to the optical inaccessibility of the mammalian liver. Here we take advantage of the transparency of the zebrafish larvae to develop a setup that allows in vivo imaging of calcium flux in zebrafish hepatocytes at cellular resolution. Using this, we provide quantitative assessment of intracellular calcium dynamics during multiple contexts, including growth, feeding, ethanol-induced stress and cell ablation. Specifically, we show that synchronized calcium oscillations are present in vivo, which are lost upon starvation. Feeding recommences calcium waves in the liver, but in a spatially restricted manner. Further, ethanol treatment as well as cell ablation induces calcium flux, but with different dynamics. The former causes asynchronous calcium oscillations, while the latter leads to a single calcium spike. Overall, we demonstrate the presence of oscillations, waves and spikes in vivo. Thus, our study introduces a platform for observing diverse calcium dynamics while maintaining the native environment of the liver, which will help investigations into the dissection of molecular mechanisms supporting the intra- and intercellular calcium signaling in the liver.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted January 12, 2022.
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In vivo imaging of calcium dynamics in zebrafish hepatocytes
Macarena Pozo-Morales, Inés Garteizgogeascoa, Camille Perazzolo, Sumeet Pal Singh
bioRxiv 2022.01.11.475536; doi: https://doi.org/10.1101/2022.01.11.475536
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In vivo imaging of calcium dynamics in zebrafish hepatocytes
Macarena Pozo-Morales, Inés Garteizgogeascoa, Camille Perazzolo, Sumeet Pal Singh
bioRxiv 2022.01.11.475536; doi: https://doi.org/10.1101/2022.01.11.475536

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