TY - JOUR TI - An in vivo reporter for tracking lipid droplet dynamics in transparent zebrafish AU - Lumaquin, Dianne AU - Johns, Eleanor AU - Montal, Emily AU - Weiss, Joshua M AU - Ola, David AU - Abuhashem, Abderhman AU - White, Richard M A2 - Schlegel, Amnon A2 - Stainier, Didier YR A2 - Schlegel, Amnon A2 - Minchin, James VL - 10 PY - 2021 DA - 2021/06/11 SP - e64744 C1 - eLife 2021;10:e64744 DO - 10.7554/eLife.64744 UR - https://doi.org/10.7554/eLife.64744 AB - 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. KW - adipocyte KW - obesity KW - lipid droplet KW - cancer metabolism JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -