Progesterone blocks cholesterol translocation from lysosomes

J. D. Butler, J. Blanchette-Mackie, E. Goldin, R. R. O'Neill, G. Carstea, C. F. Roff, M. C. Patterson, S. Patel, M. E. Comly, A. Cooney, M. T. Vanier, R. O. Brady, P. G. Pentchev

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Abstract

Fluorescent microscopic examination of fibroblasts cultured with low density lipoprotein (LDL) and progesterone (10 μg/ml) for 24 h revealed extensive filipin-cholesterol staining of perinuclear lysosomes. Levels of unesterified cholesterol were 2-fold greater than in fibroblasts cultured with LDL alone. Progesterone strongly blocked cholesteryl ester synthesis. When cellular uptake of LDL was monitored in the presence of 58035, a specific inhibitor of acyl-CoA:cholesterol acyltransferase, excess unesterified cholesterol was not stored in lysosomes. Discontinuation of LDL uptake in conjunction with progesterone washout markedly reversed the filipin-cholesterol staining of lysosomes. Reversal of the lysosomal cholesterol lipidosis was associated with a rapid burst of cholesteryl ester synthesis and a normalization of the cellular levels of free and esterified cholesterol. In contrast to normal cells, progesterone removal from Niemann- Pick C fibroblasts did not reverse the lysosomal cholesterol accumulation of these mutant cultures. The metabolic precursor of progesterone, pregnenolone, also induced extensive accumulation of cholesterol in lysosomes. Other steroids induced less vacuolar cholesterol accumulation in the following decreasing order: corticosterone and testosterone, promegestone, RU 486. The relative inhibition of cellular cholesterol esterification by the steroids paralleled their respective abilities to sequester cholesterol in lysosomes rather than their inhibition of acyl-CoA:cholesterol acyltransferase activity in cell-free extracts. The progesterone-related inhibition and restoration of lysosomal cholesterol trafficking is a useful experimental means of studying intracellular cholesterol transport. A particularly important feature of its utility is the facile reversibility of the steroid-induced block. The lysosomal cholesterol lipidosis established with a hydrophobic amine, U18666A, was not as readily reversed.

Original languageEnglish (US)
Pages (from-to)23797-23805
Number of pages9
JournalJournal of Biological Chemistry
Volume267
Issue number33
StatePublished - 1992
Externally publishedYes

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Lysosomes
Progesterone
Cholesterol
LDL Lipoproteins
Fibroblasts
Filipin
Lipidoses
Sterol O-Acyltransferase
Cholesterol Esters
Steroids
Promegestone
Staining and Labeling
Mifepristone
Pregnenolone
Esterification
Corticosterone
Cell Extracts
Restoration
Amines
Testosterone

ASJC Scopus subject areas

  • Biochemistry

Cite this

Butler, J. D., Blanchette-Mackie, J., Goldin, E., O'Neill, R. R., Carstea, G., Roff, C. F., ... Pentchev, P. G. (1992). Progesterone blocks cholesterol translocation from lysosomes. Journal of Biological Chemistry, 267(33), 23797-23805.

Progesterone blocks cholesterol translocation from lysosomes. / Butler, J. D.; Blanchette-Mackie, J.; Goldin, E.; O'Neill, R. R.; Carstea, G.; Roff, C. F.; Patterson, M. C.; Patel, S.; Comly, M. E.; Cooney, A.; Vanier, M. T.; Brady, R. O.; Pentchev, P. G.

In: Journal of Biological Chemistry, Vol. 267, No. 33, 1992, p. 23797-23805.

Research output: Contribution to journalArticle

Butler, JD, Blanchette-Mackie, J, Goldin, E, O'Neill, RR, Carstea, G, Roff, CF, Patterson, MC, Patel, S, Comly, ME, Cooney, A, Vanier, MT, Brady, RO & Pentchev, PG 1992, 'Progesterone blocks cholesterol translocation from lysosomes', Journal of Biological Chemistry, vol. 267, no. 33, pp. 23797-23805.
Butler JD, Blanchette-Mackie J, Goldin E, O'Neill RR, Carstea G, Roff CF et al. Progesterone blocks cholesterol translocation from lysosomes. Journal of Biological Chemistry. 1992;267(33):23797-23805.
Butler, J. D. ; Blanchette-Mackie, J. ; Goldin, E. ; O'Neill, R. R. ; Carstea, G. ; Roff, C. F. ; Patterson, M. C. ; Patel, S. ; Comly, M. E. ; Cooney, A. ; Vanier, M. T. ; Brady, R. O. ; Pentchev, P. G. / Progesterone blocks cholesterol translocation from lysosomes. In: Journal of Biological Chemistry. 1992 ; Vol. 267, No. 33. pp. 23797-23805.
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T1 - Progesterone blocks cholesterol translocation from lysosomes

AU - Butler, J. D.

AU - Blanchette-Mackie, J.

AU - Goldin, E.

AU - O'Neill, R. R.

AU - Carstea, G.

AU - Roff, C. F.

AU - Patterson, M. C.

AU - Patel, S.

AU - Comly, M. E.

AU - Cooney, A.

AU - Vanier, M. T.

AU - Brady, R. O.

AU - Pentchev, P. G.

PY - 1992

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N2 - Fluorescent microscopic examination of fibroblasts cultured with low density lipoprotein (LDL) and progesterone (10 μg/ml) for 24 h revealed extensive filipin-cholesterol staining of perinuclear lysosomes. Levels of unesterified cholesterol were 2-fold greater than in fibroblasts cultured with LDL alone. Progesterone strongly blocked cholesteryl ester synthesis. When cellular uptake of LDL was monitored in the presence of 58035, a specific inhibitor of acyl-CoA:cholesterol acyltransferase, excess unesterified cholesterol was not stored in lysosomes. Discontinuation of LDL uptake in conjunction with progesterone washout markedly reversed the filipin-cholesterol staining of lysosomes. Reversal of the lysosomal cholesterol lipidosis was associated with a rapid burst of cholesteryl ester synthesis and a normalization of the cellular levels of free and esterified cholesterol. In contrast to normal cells, progesterone removal from Niemann- Pick C fibroblasts did not reverse the lysosomal cholesterol accumulation of these mutant cultures. The metabolic precursor of progesterone, pregnenolone, also induced extensive accumulation of cholesterol in lysosomes. Other steroids induced less vacuolar cholesterol accumulation in the following decreasing order: corticosterone and testosterone, promegestone, RU 486. The relative inhibition of cellular cholesterol esterification by the steroids paralleled their respective abilities to sequester cholesterol in lysosomes rather than their inhibition of acyl-CoA:cholesterol acyltransferase activity in cell-free extracts. The progesterone-related inhibition and restoration of lysosomal cholesterol trafficking is a useful experimental means of studying intracellular cholesterol transport. A particularly important feature of its utility is the facile reversibility of the steroid-induced block. The lysosomal cholesterol lipidosis established with a hydrophobic amine, U18666A, was not as readily reversed.

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