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An in vivo reporter for tracking lipid droplet dynamics in transparent zebrafish

Dianne Lumaquin, Eleanor Johns, Joshua Weiss, Emily Montal, Olayinka Ooladipupo, Abderhman Abuhashem, Richard M. White
doi: https://doi.org/10.1101/2020.11.09.375667
Dianne Lumaquin
1Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA
2Weill Cornell/Rockefeller/Sloan Kettering Tri-Institutional MD-PhD Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA
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Eleanor Johns
1Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA
3Gerstner Sloan Kettering Graduate School of Biomedical Sciences, Memorial Sloan Kettering Cancer Center, New York, NY, USA
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Joshua Weiss
1Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA
2Weill Cornell/Rockefeller/Sloan Kettering Tri-Institutional MD-PhD Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA
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Emily Montal
1Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA
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Olayinka Ooladipupo
1Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA
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Abderhman Abuhashem
2Weill Cornell/Rockefeller/Sloan Kettering Tri-Institutional MD-PhD Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA
4Developmental Biology Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA
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Richard M. White
1Cancer Biology and Genetics Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA
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  • For correspondence: whiter@mskcc.org
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Abstract

Lipid droplets are lipid storage organelles found in nearly all cell types from adipocytes to cancer cells. Although increasingly implicated in disease, current methods to study lipid droplets require fixation or static imaging which limits investigation of their rapid in vivo dynamics. To address this, we created a lipid droplet transgenic reporter in whole animals and cell culture by fusing tdTOMATO to Perilipin-2 (PLIN2), a lipid droplet structural protein. Expression of this transgene in transparent casper zebrafish enabled in vivo imaging of adipose depots responsive to nutrient deprivation and high-fat diet. Using this system, we tested novel regulators of lipolysis, revealing an unexpected role for nitric oxide in modulating adipocyte lipid droplets. Similarly, we expressed the PLIN2-tdTOMATO transgene in melanoma cells and found that the nitric oxide pathway also regulated lipid droplets in cancer. This model offers a tractable imaging platform to study lipid droplets across cell types and disease contexts.

Competing Interest Statement

D.L., E.J., J.W., E.M., O.O., and A.A. do not have competing interests to declare. R.M.W is a paid consultant to N-of-One Therapeutics, a subsidiary of Qiagen. R.M.W is on the scientific advisory board of Consano, but receives no income for this. R.M.W receives royalty payments for the use of the casper zebrafish line from Carolina Biologicals.

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-NC 4.0 International license.
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Posted November 09, 2020.
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An in vivo reporter for tracking lipid droplet dynamics in transparent zebrafish
Dianne Lumaquin, Eleanor Johns, Joshua Weiss, Emily Montal, Olayinka Ooladipupo, Abderhman Abuhashem, Richard M. White
bioRxiv 2020.11.09.375667; doi: https://doi.org/10.1101/2020.11.09.375667
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An in vivo reporter for tracking lipid droplet dynamics in transparent zebrafish
Dianne Lumaquin, Eleanor Johns, Joshua Weiss, Emily Montal, Olayinka Ooladipupo, Abderhman Abuhashem, Richard M. White
bioRxiv 2020.11.09.375667; doi: https://doi.org/10.1101/2020.11.09.375667

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