Abstract
Acid sphingomyelinase-deficient (asmase−/−) mice generated by gene targeting abundantly store sphingomyelin in the reticuloendothelial system of liver, spleen, bone marrow, and in brain. Liver cells of asmase−/− mice accumulate sphingomyelin and glycosphingolipids in purified lipid bilayers of microsomes, Golgi, and the plasma membrane, but cholesterol is depleted in the plasma membrane. Detergent-insoluble glycolipid-enriched membrane microdomains (GEM) can be isolated from hepatocytes, embryonic fibroblasts, and splenocytes of wild-type, but not of asmase−/− mice, by sucrose gradient density centrifugation. Lck and other Src-family kinases are reduced in isopycnic fractions of asmase−/− splenocytes compared to GEM-containing fractions of wild-type cells. The proliferation of asmase−/− T lymphocytes is reduced, whereas their susceptibility to Fas-induced apoptosis is increased after T cell receptor (TCR) stimulation. TNF receptor I signaling remains unimpaired. The perturbation of GEM impairs tyrosine phosphorylation and, consequently, mitogenic signaling of the TCR. Reduced MAPK activity-dependent FLICE-like inhibitory protein (FLIP) expression in asmase−/− T lymphocytes increases their sensitivity towards Fas-mediated apoptosis. Cell Death and Differentiation (2000) 7, 413–424
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Abbreviations
- AICD:
-
activation-induced cell death
- aSMase:
-
acid sphingomyelinase
- CHX:
-
cycloheximide
- DISC:
-
death-inducing signaling complex
- FLIP:
-
FLICE-like inhibitory protein
- GEM:
-
glycolipid-enriched membrane microdomains
- SPM:
-
sphingomyelin
- PMA:
-
phorbol myristoyl acetate
- TNF-α:
-
tumor necrosis factor-α
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Acknowledgements
We thank B Jenke for her excellent technical assistance and P Bauer for her kind help with FACS analyses. This work was supported by the Deutsche Forschungsgemeinschaft SFB 243, Project A4, and by the Federal Ministry for Education, Science, Research and Technology 01KS9502 (Center for Molecular Medicine Cologne, ZMMK), Project 24.
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Nix, M., Stoffel, W. Perturbation of membrane microdomains reduces mitogenic signaling and increases susceptibility to apoptosis after T cell receptor stimulation. Cell Death Differ 7, 413–424 (2000). https://doi.org/10.1038/sj.cdd.4400666
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DOI: https://doi.org/10.1038/sj.cdd.4400666
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