PUBLICATION

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

Authors
Lumaquin, D., Johns, E., Montal, E., Weiss, J.M., Ola, D., Abuhashem, A., White, R.M.
ID
ZDB-PUB-210612-5
Date
2021
Source
eLIFE   10: (Journal)
Registered Authors
White, Richard M.
Keywords
adipocyte, cancer metabolism, cell biology, lipid droplet, obesity, zebrafish
MeSH Terms
  • Adipocytes/metabolism*
  • Adipose Tissue/metabolism
  • Animals
  • Cell Line, Tumor
  • Diet, High-Fat
  • Lipid Droplets/metabolism*
  • Lipid Metabolism
  • Lipolysis
  • Luminescent Proteins/metabolism*
  • Melanoma/metabolism
  • Nitric Oxide/metabolism
  • Perilipin-2/metabolism*
  • Phenylurea Compounds/pharmacology
  • Zebrafish/metabolism
  • Zebrafish Proteins/metabolism*
PubMed
34114952 Full text @ Elife
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 in vertebrate models rely on static imaging or the use of fluorescent dyes, limiting 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. Simultaneously, we performed a large-scale in vitro chemical screen of 1280 compounds and identified several novel regulators of lipolysis in adipocytes. Using our Tg(-3.5ubb:plin2-tdTomato) zebrafish line, we validated several of these novel regulators and revealed 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 using chemical, dietary, or genetic perturbations.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping