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Journal of Lipid Research, Vol 30, 403-413, Copyright © 1989 by Lipid Research, Inc.
Quantification and regulation of apolipoprotein E expression in rat Kupffer cells
PA Dawson, LM Lukaszewski, PF Ells, CC Malbon and DL Williams
Department of Pharmacological Sciences, State University of New York, Stony Brook 11794-8651.
Apolipoprotein E (apoE) is synthesized by a wide variety of cells including
cells of the monocyte-macrophage lineage. In order to assess the
quantitative significance of apoE synthesis in a mature tissue macrophage,
apoE synthesis was compared in Kupffer cells and hepatocytes isolated from
rat liver. Immunoreactive apoE synthesized by both cell types exhibited
identical isoform patterns when examined by high-resolution two-dimensional
gel analysis. ApoE synthesis was not detected in hepatic endothelial cells.
Northern blot analysis using a rat apoE cDNA probe demonstrated a single
mRNA species of approximately 1200 nucleotides in freshly isolated
hepatocytes and Kupffer cells. The absolute content of apoE mRNA in each
cell type was determined with a DNA-excess solution hybridization assay.
The apoE mRNA content (pg/microgram RNA) for Kupffer cells and hepatocytes
was 35.7 and 98.8, respectively. Accounting for cellular RNA content and
the population size of each cell type in the liver, Kupffer cells were
calculated to contain about 0.7% of liver apoE mRNA; hepatocytes account
almost quantitatively for the remainder. These results suggest that Kupffer
cells are not major contributors to the plasma apoE pool. After intravenous
injection of bacterial endotoxin, apoE mRNA was decreased in freshly
isolated Kupffer cells whereas whole liver showed no change in apoE mRNA.
Endotoxin treatment had no effect on the apoE mRNA content in several
peripheral tissues. These results indicate that apoE expression in vivo is
differentially regulated by endotoxin in Kupffer cells as compared to
hepatocytes or apoE-producing cells in peripheral tissues.

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Copyright © 1989 by the American Society for Biochemistry and Molecular Biology.
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