Endocytosed β-VLDL and LDL are delivered to different intracellular vesicles in mouse peritoneal macrophages

Ira Tabas, Sungtae Lim, Xiangxi Xu, Frederick R. Maxfield

Research output: Contribution to journalArticle

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Abstract

Hypercholesterolemic rabbit β-VLDL and human LDL are both internalized by mouse peritoneal macrophages by receptor-mediated endocytosis. However, only β-VLDL (which binds to the cells with a much higher affinity than LDL) markedly stimulates acyl-CoA/cholesterol acyl transferase (ACAT) and induces foam cell formation in these cells. As an initial step to test whether the two lipoproteins might be targeted to different organelles (which might differ in their ability to deliver cholesterol to microsomal ACAT), we studied the endocytic pathways of β-VLDL and LDL. Lipoproteins were labeled with the non-transferable fluorescent label, DiI. When the macrophages were incubated with DiI-LDL for 10 min at 37°C, the fluorescence was concentrated near the center of the cell both in heavily labeled vesicles and in a diffuse pattern. The pattern with DiI-β-VLDL was quite different: an array of bright vesicles throughout the cytoplasm was the predominant feature. Differences in distribution were seen as early as 2 min of incubation and persisted throughout a 10-min chase period. By using a procedure in which photobleaching of DiI fluorescence converts diaminobenzidine into an electron-dense marker, we were able to identify at the ultrastructural level vesicles containing electron-dense material in cells incubated with DiI-β-VLDL. Human E2/E2 β-VLDL (from a patient with familial dysbetalipoproteinemia), which has a binding affinity and ACAT-stimulatory potential similar to LDL, gave a pattern of fluorescence virtually identical to LDL. Pulse-chase studies with 125I-labeled and [3H]cholesteryl ester-labeled lipoproteins disclosed that both protein degradation and cholesteryl ester hydrolysis were markedly retarded in β-VLDL compared with LDL. Thus, in mouse peritoneal macrophages, endocytosed β-VLDL appears in a distinct set of widely-distributed vesicles not seen with LDL (or with E2-β-VLDL) and, compared with LDL, has a markedly diminished rate of protein degradation and cholesteryl ester hydrolysis. The differential routing of LDL and β-VLDL may provide a mechanism for differences in ACAT-stimulatory potential between the two lipoproteins.

Original languageEnglish
Pages (from-to)929-940
Number of pages12
JournalJournal of Cell Biology
Volume111
Issue number3
StatePublished - Sep 1 1990
Externally publishedYes

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Peritoneal Macrophages
Endocytosis
Acyl Coenzyme A
Transferases
Cholesterol
Lipoproteins
Cholesterol Esters
Fluorescence
Proteolysis
Hydrolysis
Hyperlipoproteinemia Type III
oxidized low density lipoprotein
Electrons
Photobleaching
Foam Cells
Exercise Test
Organelles
Cytoplasm
Macrophages
Rabbits

ASJC Scopus subject areas

  • Cell Biology

Cite this

Endocytosed β-VLDL and LDL are delivered to different intracellular vesicles in mouse peritoneal macrophages. / Tabas, Ira; Lim, Sungtae; Xu, Xiangxi; Maxfield, Frederick R.

In: Journal of Cell Biology, Vol. 111, No. 3, 01.09.1990, p. 929-940.

Research output: Contribution to journalArticle

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