Abstract
Increased brain ganglioside levels are a hallmark of various neuroinflammatory pathologies. Here, we provide evidence that murine microglia can secrete disialoganglioside GD3 upon exposure to inflammatory stimuli. Comparison of different neural cell types revealed a particular and specific sensitivity of oligodendrocytes towards exogenous GD3. Oligodendrocyte death triggered by GD3 was preceded by degeneration of cellular processes, and associated with typical features of apoptosis, such as chromatin condensation, exposure of phosphatidylserine, release of cytochrome c from mitochondria, and loss of mitochondrial membrane potential, followed by the loss of plasma membrane integrity and detachment of disintegrated oligodendrocytes. Overexpression of bcl-2 partially protected oligodendrocytes from death. In contrast, treatment with the pan-caspase inhibitor zVAD-fmk did not prevent phosphatidylserine exposure, chromatin margination at the nuclear periphery, and death, although caspase-3 was blocked. Thus, GD3 produced by microglia under neuroinflammatory conditions may function as a novel mediator triggering mitochondria-mediated, but caspase-independent, apoptosis-like death of oligodendrocytes.
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Abbreviations
- CSF:
-
cerebrospinal fluid
- CGC:
-
cerebellar granule neurons
- CNPase:
-
cyclic nucleotide phosphodiesterase
- cyt c:
-
cytochrome c
- EH-1:
-
ethidium homodimer-1
- FCS:
-
foetal calf serum
- GD3:
-
disialoganglioside 3
- GD1a:
-
disialoganglioside 1a
- GD1b:
-
disialoganglioside 1b
- GM3:
-
monosialoganglioside 3
- LPS:
-
lipopolysaccharide
- MOG:
-
myelin oligodendrocyte glycoprotein
- MPT:
-
mitochondrial permeability transition
- MTT:
-
3-(4,5-Dimethyldiazol-2-yl)-3,5-diphenyltetrazolium bromide
- MS:
-
Multiple Sclerosis
- NO:
-
nitric oxide
- PS:
-
phosphatidylserine
- P/S:
-
penicillin/streptomycine
- TLC:
-
thin layer chromatography
- TMRE:
-
tetramethylrhodamine-ethylester
- TNFα:
-
tumour necrosis factor alpha
- ΔΨm:
-
mitochondrial membrane potential
- zVAD-fmk:
-
N-benzyloxycarbonyl-Val-Ala-aspartyl-fluoromethylketone
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Acknowledgements
The excellent technical assistance of Elvira Gawlitta-Gorka is gratefully acknowledged. We are grateful to Dr. J Schenkel (Institute of Physiology II, University of Heidelberg, Germany) and Dr. R Jäger (Forschungszentrum Karlsruhe, Germany) for providing bcl-2 transgenic mice, to Dr. T Mürdter (Dr. M Fischer-Bosch-Institute for Clinical Pharmacology, Stuttgart, Germany) for measurements of ceramides by combined gas-chromatography and mass-spectroscopy and to Dr. J Castro-Palomino (Bayer AG, Wuppertal, Germany) and Prof. Dr. RR Schmidt (University of Konstanz, Germany) for chemical synthesis of GD3. This study was supported by the DFG grant We686/18, by AIRC, by ISS Progetto Sclerosi Multiple as well as the EEC grants BMH4CT97-2410 and 12029-97-06 F1ED ISP D. F Malisan was supported by a fellowship from FIRC.
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Simon, B., Malisan, F., Testi, R. et al. Disialoganglioside GD3 is released by microglia and induces oligodendrocyte apoptosis. Cell Death Differ 9, 758–767 (2002). https://doi.org/10.1038/sj.cdd.4401027
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DOI: https://doi.org/10.1038/sj.cdd.4401027
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