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.


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.
Boström
Pontus
P
Wallenberg Laboratory for Cardiovascular Research, Göteborg University, Göteborg, Sweden.
Rutberg
Mikael
M
Ericsson
Johanna
J
Holmdahl
Peter
P
Andersson
Linda
L
Frohman
Michael A
MA
Borén
Jan
J
Olofsson
Sven-Olof
SO
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D050197:Atherosclerosis / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins / Q000494:pharmacology; D050356:Lipid Metabolism; D008565:Membrane Proteins / Q000494:pharmacology; D051379:Mice; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism*; D020409:Molecular Motor Proteins / Q000378:metabolism; D041681:NIH 3T3 Cells; D015739:Nocodazole / Q000494:pharmacology; D019301:Oleic Acid / Q000737:chemistry / Q000493:pharmacokinetics*; D010078:Oxazines; D010316:Particle Size; D000070780:Perilipin-2; D014280:Triglycerides / Q000096:biosynthesis
D000970:Antineoplastic Agents; D005456:Fluorescent Dyes; D008565:Membrane Proteins; D020409:Molecular Motor Proteins; D010078:Oxazines; D000070780:Perilipin-2; C000602983:Plin2 protein, mouse; D014280:Triglycerides; D049452:Green Fluorescent Proteins; D019301:Oleic Acid; C044808:nile red; D015739:Nocodazole