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Journal of Lipid Research, Vol 36, 1211-1226, Copyright © 1995 by Lipid Research, Inc.
EJ Blanchette-Mackie, NK Dwyer, T Barber, RA Coxey, T Takeda, CM Rondinone, JL Theodorakis, AS Greenberg and C Londos
Immunocytochemistry was used to determine the intracellular location of
perilipins in adipocytes and the occurrence of these proteins in tissues
involved in triacylglycerol metabolism. Confocal microscopy and
3-dimensional analysis of 3T3-L1 adipocytes showed that perilipin
immunofluorescence, present on the surfaces of all sized lipid droplets,
appeared unevenly dispersed on the surfaces of many large lipid droplets.
Electron microscopy revealed that immunogold staining for perilipin was
located directly on the surface layer apposed to and surrounding the core
triacylglycerol of intracellular lipid droplets of adipocytes in culture or
from white and brown adipose tissue. Freeze- fracture electron microscopy
indicated that the hydrophobic face of this surface monolayer contained
particles identical in size and distribution to intramembranous particles
(IMPs), which are unique structural features of the hydrophobic faces of
bilayered membranes. Also, freeze-fracture replicas revealed areas of
continuity between the surface layer of lipid droplets and the membrane
leaflets of endoplasmic reticulum, suggesting that the droplet monolayer
surface is an area of endoplasmic reticulum membrane leaflet modified by
its unique content of perilipin. Microperoxisomes, identified by
immunostaining for catalase, were found closely associated with lipid
droplets, but external to and not in contact with the lipid droplet surface
layer. Vimentin, identified by immunofluorescence, was present around the
periphery of most lipid droplets in 3T3-L1 cells during early stages of
adipocyte development but, in contrast to perilipins, vimentin was not
around the periphery of many large lipid droplets in mature cells. Although
perilipin was at the surface of lipid droplets in adipocytes of lactating
mammary gland, none was found to be associated with the milk lipid droplets
in alveolar epithelial cells, nor was the protein found on the surfaces of
lipid droplets in hepatocytes. Studies in mammary gland show that perilipin
immunostaining will be a valuable tool for the identification of tissue
adipocytes severely depleted of their triacylglycerol stores and thus
without their characteristic spherical shape. Perilipin's singular location
on the surface monolayer of intracellular lipid droplets supports an
intimate role for the protein in the triacylglycerol metabolic functions of
adipocytes.
ARTICLES
Perilipin is located on the surface layer of intracellular lipid droplets in adipocytes
Lipid Cell Biology Section, National Institutes of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0850, USA.
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