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Journal of Lipid Research, Vol 31, 1589-1603, Copyright © 1990 by Lipid Research, Inc.
RL Hamilton, JS Wong, LS Guo, S Krisans and RJ Havel
The distribution of apolipoprotein (apo) E in rat hepatocytes was
investigated with an affinity-purified polyclonal antibody raised against
apoE isolated from hepatogeneous very low density lipoproteins (VLDL). The
distribution of this antibody was visualized with colloidal gold complexed
to anti-rabbit IgG. By epipolarization microscopy, apoE was found uniformly
along the basolateral surfaces of all hepatic parenchymal cells, showing a
striking intensity along the sinusoidal front. Punctate deposits of
colloidal gold appeared to be randomly distributed within all hepatocytes.
Widely scattered Kupffer cells also stained for apoE. Electron microscopic
examination of immunogold- labeled cryothin sections showed that
hepatocytic microvilli projecting into the space of Disse consistently
contained clusters of immunogold. The gold particles were variably
associated with evident lipoprotein particles, raising the possibility that
apoE alone may bind to receptors or other macromolecules at the surface of
hepatocytes. Endosomes near the sinusoidal front and multivesicular bodies
in the Golgi/biliary area labeled intensely for apoE, consistent with a
high content of apoE associated with triglyceride-rich lipoprotein remnants
contained within these organelles. Some but not all nascent VLDL particles
within putative forming Golgi secretory vesicles were labeled, but many
other Golgi vesicles and cisternae that lacked evident VLDL particles were
also labeled. These results suggest that at least some apoE associates with
nascent VLDL in forming Golgi secretory vesicles. Unexpectedly, the matrix
of all hepatocytic peroxisomes was heavily labeled. Immunoblots with the
affinity-purified anti-rat apoE IgG against proteins from highly purified
peroxisomes isolated from rat hepatocytes revealed a protein with an
apparent molecular mass of 34.5 kDa, similar to that of rat apoE in rat
blood plasma. In addition, gold was sometimes found in the area either
adjacent to peroxisomes or between multivesicular bodies and the bile
canaliculus not evidently associated with a membranous compartment. These
observations suggest that apoE may participate in interorganellar
cholesterol transport within hepatocytes.
ARTICLES
Apolipoprotein E localization in rat hepatocytes by immunogold labeling of cryothin sections
Department of Anatomy, University of California, San Francisco 94143- 0130.
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