Cytosolic lipid droplets increase in size by microtubule-dependent complex formation

• OBJECTIVE: Adipocyte differentiation-related protein (ADRP)-containing lipid droplets have an essential role in the development of insulin resistance and atherosclerosis. Such droplets form in a cell-free system with a diameter of 0.1 to 0.4 microm, while the droplets present in cells vary in size, from small to very large, suggesting that the droplets can increase in size after being assembled. We have addressed this possibility. METHODS AND RESULTS: Experiments in NIH 3T3 cells demonstrated that the lipid droplets could increase in size independently of triglyceride biosynthesis. NIH 3T3 cells were either microinjected with ADRP-GFP (green fluorescent protein) or stained with Nile Red and followed by confocal microscopy and time-lapse recordings. The results showed that lipid droplets formed complexes with each other, with a volume equal to the sum of the merging particles. The formation of complexes could be inhibited by the nocodazole-induced depolymerization of the microtubules; thus, the process is dependent on microtubules. The presence of dynein on ADRP-containing droplets supports a role for this motor protein. CONCLUSIONS: Lipid droplets can grow after they have been assembled. This increase in size is independent of triglyceride biosynthesis and involves formation of complexes, which requires intact microtubules.

Nyckelord

Animals

Antineoplastic Agents/pharmacology

Atherosclerosis/metabolism

Cytosol/metabolism

Fluorescent Dyes

Green Fluorescent Proteins/pharmacology

*Lipid Metabolism

Membrane Proteins/pharmacology

Mice

Microtubules/drug effects/*metabolism

Molecular Motor Proteins/metabolism

NIH 3T3 Cells

Nocodazole/pharmacology

Oleic Acid/chemistry/*pharmacokinetics

Oxazines

Particle Size

Triglycerides/biosynthesis

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