Abstract
Lipid droplets are key organelles in energy homeostasis. LD coating proteins, the Perilipins, are strongly involved in lipid metabolism and metabolic disorders. Abundance of Perilipin 5 (PLIN5), a LD coating protein exclusively found in oxidative tissue, increases lipid content in skeletal muscle without negative metabolic effects. In this work, we use quantitative, label-free coherent anti-Stokes Raman scattering (CARS) microscopy to analyze lipid composition in LDs in muscle upon PLIN5 overexpression in vivo and in vitro. Additionally, single-band vibrational microscopy allows live-cell imaging of lipids in vitro. Combined with fluorescence microscopy, the effect of PLIN5 on lipid dynamics under conditions of lipid surplus and after lipolytic stimulation were observed. Apart from confirming the previously reported size increase of LDs, vibrational imaging also revealed that PLIN5 has marked effects on the local lipid density and composition in droplets, and alters lipid dynamics under varying conditions...
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