Inhibition of [3h]thymidine transport is a nonspecific effect of pdmp in primary cultures of mouse epidermal keratinocytes

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Inhibitors of sphingolipid metabolism are frequently used to investigate the role of ceramide and other sphingolipids as intracellular signaling molecules. For example, the inhibitor of glucosylceramide synthase D-threo-1-phenyl-2-decanoylamino-3-morpholino-l-propanol (PDMP) is commonly used to deplete glycosphingolipids and increase ceramide levels. Ceramide is known to induce growth arrest and differentiation of keratinocytes, and we hypothesized that PDMP would increase ceramide levels and induce growth arrest of primary cultures of mouse epidermal keratinocytes. As expected, PDMP increased ceramide levels and decreased the incorporation of [3H]thymidine into DNA, but surprisingly, PDMP was found to rapidly inhibit the intracellular transport of [3H]thymidine. This is likely due to a direct effect on nucleoside transport by PDMP and not due to elevations in ceramide levels because increasing ceramide levels by the addition of exogenous sphingomyelinase had no effect on [3H]thymidine transport. Furthermore, it is unlikely that the decreased [3H]thymidine transport is in response to growth arrest because PDMP had no effect on the cell cycle profile of keratinocytes. These results reveal that PDMP inhibits nucleoside transport independent of effects on ceramide levels or cell growth but probably by a direct effect on the nucleoside transport apparatus. Thus, this compound may be unsuitable for investigating the role of ceramide or other sphingolipids in some cellular processes.

Original languageEnglish (US)
Pages (from-to)1219-1224
Number of pages6
JournalJournal of Pharmacology and Experimental Therapeutics
Volume294
Issue number3
StatePublished - Sep 13 2000

Fingerprint

Ceramides
Keratinocytes
Thymidine
Sphingolipids
Nucleosides
ceramide glucosyltransferase
Growth
Sphingomyelin Phosphodiesterase
1-Propanol
Morpholinos
Glycogen Synthase
Glycosphingolipids
RV 538
Cell Cycle
DNA

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology

Cite this

@article{bbdc237c034d485d96be91df382c9c43,
title = "Inhibition of [3h]thymidine transport is a nonspecific effect of pdmp in primary cultures of mouse epidermal keratinocytes",
abstract = "Inhibitors of sphingolipid metabolism are frequently used to investigate the role of ceramide and other sphingolipids as intracellular signaling molecules. For example, the inhibitor of glucosylceramide synthase D-threo-1-phenyl-2-decanoylamino-3-morpholino-l-propanol (PDMP) is commonly used to deplete glycosphingolipids and increase ceramide levels. Ceramide is known to induce growth arrest and differentiation of keratinocytes, and we hypothesized that PDMP would increase ceramide levels and induce growth arrest of primary cultures of mouse epidermal keratinocytes. As expected, PDMP increased ceramide levels and decreased the incorporation of [3H]thymidine into DNA, but surprisingly, PDMP was found to rapidly inhibit the intracellular transport of [3H]thymidine. This is likely due to a direct effect on nucleoside transport by PDMP and not due to elevations in ceramide levels because increasing ceramide levels by the addition of exogenous sphingomyelinase had no effect on [3H]thymidine transport. Furthermore, it is unlikely that the decreased [3H]thymidine transport is in response to growth arrest because PDMP had no effect on the cell cycle profile of keratinocytes. These results reveal that PDMP inhibits nucleoside transport independent of effects on ceramide levels or cell growth but probably by a direct effect on the nucleoside transport apparatus. Thus, this compound may be unsuitable for investigating the role of ceramide or other sphingolipids in some cellular processes.",
author = "Griner, {R. D.} and Bollag, {W. B.}",
year = "2000",
month = "9",
day = "13",
language = "English (US)",
volume = "294",
pages = "1219--1224",
journal = "The Journal of pharmacology and experimental therapeutics",
issn = "0022-3565",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "3",

}

TY - JOUR

T1 - Inhibition of [3h]thymidine transport is a nonspecific effect of pdmp in primary cultures of mouse epidermal keratinocytes

AU - Griner, R. D.

AU - Bollag, W. B.

PY - 2000/9/13

Y1 - 2000/9/13

N2 - Inhibitors of sphingolipid metabolism are frequently used to investigate the role of ceramide and other sphingolipids as intracellular signaling molecules. For example, the inhibitor of glucosylceramide synthase D-threo-1-phenyl-2-decanoylamino-3-morpholino-l-propanol (PDMP) is commonly used to deplete glycosphingolipids and increase ceramide levels. Ceramide is known to induce growth arrest and differentiation of keratinocytes, and we hypothesized that PDMP would increase ceramide levels and induce growth arrest of primary cultures of mouse epidermal keratinocytes. As expected, PDMP increased ceramide levels and decreased the incorporation of [3H]thymidine into DNA, but surprisingly, PDMP was found to rapidly inhibit the intracellular transport of [3H]thymidine. This is likely due to a direct effect on nucleoside transport by PDMP and not due to elevations in ceramide levels because increasing ceramide levels by the addition of exogenous sphingomyelinase had no effect on [3H]thymidine transport. Furthermore, it is unlikely that the decreased [3H]thymidine transport is in response to growth arrest because PDMP had no effect on the cell cycle profile of keratinocytes. These results reveal that PDMP inhibits nucleoside transport independent of effects on ceramide levels or cell growth but probably by a direct effect on the nucleoside transport apparatus. Thus, this compound may be unsuitable for investigating the role of ceramide or other sphingolipids in some cellular processes.

AB - Inhibitors of sphingolipid metabolism are frequently used to investigate the role of ceramide and other sphingolipids as intracellular signaling molecules. For example, the inhibitor of glucosylceramide synthase D-threo-1-phenyl-2-decanoylamino-3-morpholino-l-propanol (PDMP) is commonly used to deplete glycosphingolipids and increase ceramide levels. Ceramide is known to induce growth arrest and differentiation of keratinocytes, and we hypothesized that PDMP would increase ceramide levels and induce growth arrest of primary cultures of mouse epidermal keratinocytes. As expected, PDMP increased ceramide levels and decreased the incorporation of [3H]thymidine into DNA, but surprisingly, PDMP was found to rapidly inhibit the intracellular transport of [3H]thymidine. This is likely due to a direct effect on nucleoside transport by PDMP and not due to elevations in ceramide levels because increasing ceramide levels by the addition of exogenous sphingomyelinase had no effect on [3H]thymidine transport. Furthermore, it is unlikely that the decreased [3H]thymidine transport is in response to growth arrest because PDMP had no effect on the cell cycle profile of keratinocytes. These results reveal that PDMP inhibits nucleoside transport independent of effects on ceramide levels or cell growth but probably by a direct effect on the nucleoside transport apparatus. Thus, this compound may be unsuitable for investigating the role of ceramide or other sphingolipids in some cellular processes.

UR - http://www.scopus.com/inward/record.url?scp=0033842668&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033842668&partnerID=8YFLogxK

M3 - Article

C2 - 10945880

AN - SCOPUS:0033842668

VL - 294

SP - 1219

EP - 1224

JO - The Journal of pharmacology and experimental therapeutics

JF - The Journal of pharmacology and experimental therapeutics

SN - 0022-3565

IS - 3

ER -