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Journal of Lipid Research, Vol 34, 1165-1176, Copyright © 1993 by Lipid Research, Inc.
RA Coxey, PG Pentchev, G Campbell and EJ Blanchette-Mackie
Cholesterol accumulation in the Golgi of normal and Niemann-Pick Type C
(NP-C) fibroblasts was shown by freeze-fracture electron microscopy using
filipin as a probe for unesterified cholesterol. The specific distribution
of cholesterol within individual Golgi compartments could be examined
because membrane cholesterol forms complexes with filipin that are visible
as membrane deformations (pits and protuberances) in freeze-fracture
replicas. The density of filipin-cholesterol deformations, quantitated for
cis, medial, and trans Golgi cisternae and trans Golgi vacuoles, was shown
to increase in a cis to trans direction. After addition of low density
lipoproteins (LDL) to cultured fibroblasts for 24 h, the cholesterol
content increased within specific compartments of the Golgi. Normal cells
showed an increase in filipin- cholesterol deformations in membranes of
cis/medial cisternae and trans Golgi vacuoles, whereas NP-C cells showed
only an increase in membranes of trans Golgi cisternae. LDL uptake by cells
appears to induce a disparate cholesterol enrichment of Golgi compartments
of normal and mutant cells. The ability of cells to process endocytosed
cholesterol may in part depend on modulation of cholesterol-enriched
membrane transport through the Golgi, a function which appears to be
defective in NP-C cells.
ARTICLES
Differential accumulation of cholesterol in Golgi compartments of normal and Niemann-Pick type C fibroblasts incubated with LDL: a cytochemical freeze-fracture study
Endocrinology Section, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892.
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