Journal of Molecular Neuroscience

, Volume 15, Issue 2, pp 85–97 | Cite as

Pivotal role for acidic sphingomyelinase in cerebral ischemia-induced ceramide and cytokine production, and neuronal apoptosis

  • Zai Fang Yu
  • Mariana Nikolova-Karakashian
  • Daohong Zhou
  • Guanjun Cheng
  • Edward H. Schuchman
  • Mark P. Mattson


Stroke is a major cause of long-term disability, the severity of which is directly related to the numbers of neurons that succumb to the ischemic insult. The signaling cascades activated by cerebral ischemia that may either promote or protect against neuronal death are not well-understood. One injury-responsive signaling pathway that has recently been characterized in studies of non-neural cells involves cleavage of membrane sphingomyelin by acidic and/or neutral sphingomyelinase (ASMase) resulting in generation of the second messenger ceramide. We now report that transient focal cerebral ischemia induces large increases in ASMase activity, ceramide levels, and production of inflammatory cytokines in wild-type mice, but not in mice lacking ASMase. The extent of brain tissue damage is decreased and behavioral outcome improved in mice lacking ASMase. Neurons lacking ASMase exhibit decreased vulnerability to excitotoxicity and hypoxia, which is associated with decreased levels of intracellular calcium and oxyradicals. Treatment of mice with a drug that inhibits ASMase activity and ceramide production reduces ischemic neuronal injury and improves behavioral outcome, suggesting that drugs that inhibit this signaling pathway may prove beneficial in stroke patients.

Index Entries

Apoptosis calcium excitotoxicity interleukin stroke tumor necrosis factor 


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  1. Bajjalieh S. and Batchelor R. (2000) Ceramide kinase. Methods Enzymol. 311, 207–215.PubMedGoogle Scholar
  2. Ballou L. R., Laulederkind S. J., Rosloniec E. F., and Raghow R. (1996) Ceramide signalling and the immune response. Biochim. Biophys. Acta 1301, 273–287.PubMedGoogle Scholar
  3. Barone F. C., Arvin B., White R. F., Miller A., Webb C. L., Willette R. N., et al. (1997) Tumor necrosis factor-alpha. A mediator of focal ischemic brain injury. Stroke 28, 1233–1244.PubMedGoogle Scholar
  4. Bruce A. J., Boling W., Kindy M. S., Peschon J., Kraemer P. J., Carpenter M. K., et al. (1996) Altered neuronal and microglial responses to excitotoxic and ischemic brain injury in mice lacking TNF receptors. Nature Med. 2, 788–794.PubMedCrossRefGoogle Scholar
  5. Brugg B., Michel P. P., Agid Y., and Ruberg M. (1996) Ceramide induces apoptosis in cultured mesencephalic neurons. J. Neurochem. 66, 733–739.PubMedCrossRefGoogle Scholar
  6. Chen J., Nikolova-Karakashian M., Merrill A. H., and Morgan E. T. (1995) Regulation of cytochrome p450 2C11 (CYP2C11) gene expression by interleukin-1, sphingomyelin hydrolysis, and ceramides in rat hepatocytes. J. Biol. Chem. 270, 25,233–25,236.Google Scholar
  7. Cifone M. G., Roncaioli P., De Maria R., Camarda G., Santoni A., Ruberti G., and Testi R. (1995) Multiple pathways originate at the Fas/APO-1 (CD95) receptor: sequential involvement of phosphatidylcholine-specific phospholipase C and acidic sphingomyelinase in the propagation of the apoptotic signal. EMBO J. 14, 5859–5868.PubMedGoogle Scholar
  8. Cifone M. G., Migliorati G., Parroni R., Marchetti C., Millimaggi D., Santoni A., and Riccardi C. (1999) Dexamethasone-induced thymocyte apoptosis: apoptotic signal involves the sequential activation of phosphoinositide-specific phospholipase C, acidic sphingomyelinase, and caspases. Blood 93, 2282–2296.PubMedGoogle Scholar
  9. Crumrine R. C., Thomas A. L., and Morgan P. F. (1994) Attenuation of p53 expression protects against focal ischemic damage in transgenic mice. J. Cereb. Blood Flow Metab. 14, 887–891.PubMedGoogle Scholar
  10. Degli Esposti M. and McLennan H. (1998) Mitochondria and cells produce reactive oxygen species in virtual anaerobiosis: relevance to ceramide-induced apoptosis. FEBS Lett. 430, 338–342.PubMedCrossRefGoogle Scholar
  11. DeGraba T. J. (1998) The role of inflammation after acute stroke: utility of pursuing anti-adhesion molecule therapy. Neurology 51, S62-S68.PubMedGoogle Scholar
  12. Dirnagl U., Iadecola C. and Moskowitz M. A. (1999) Pathobiology of ischaemic stroke: an integrated view. Trends Neurosci. 22, 391–397.PubMedCrossRefGoogle Scholar
  13. Endres M., Fink K., Zhu J., Stagliano N. E., Bondada V., Geddes J. W., et al. (1999) Neuroprotective effects of gelsolin during murine stroke. J. Clin. Invest. 103, 347–354.PubMedCrossRefGoogle Scholar
  14. Fedoroff S., Berezovskaya O., and Maysinger D. (1997) Role of colony stimulating factor-1 in brain damage caused by ischemia. Neurosci. Biobehav. Rev. 21, 187–191.PubMedCrossRefGoogle Scholar
  15. Fink K., Zhu J., Namura S., Shimizu-Sasamata M., Endres M., Ma J., et al. (1998) Prolonged therapeutic window for ischemic brain damage caused by delayed caspase activation. J. Cereb. Blood Flow Metab. 18, 1071–1076.PubMedCrossRefGoogle Scholar
  16. France-Lanord V., Brugg B., Michel P. P., Agid Y., and Ruberg M. (1997) Mitochondrial free radical signal in ceramide-dependent apoptosis: a putative mechanism for neuronal death in Parkinson’s disease. J. Neurochem. 69, 1612–1621.PubMedCrossRefGoogle Scholar
  17. Garcia-Ruiz C., Colell A., Mari M., Morales A., and Fernandez-Checa J. C. (1997) Direct effect of ceramide on the mitochondrial electron transport chain leads to generation of reactive oxygen species. Role of mitochondrial glutathione. J. Biol. Chem. 272, 11,369–11,377.Google Scholar
  18. Gary D. S., Bruce-Keller A. J., Kindy M. S., and Mattson M. P. (1998) Ischemic and excitotoxic brain injury is enhanced in mice lacking the p55 tumor necrosis factor receptor. J. Cereb. Blood Flow Metab. 18, 1283–1287.PubMedCrossRefGoogle Scholar
  19. Goodman Y. and Mattson M. P. (1996) Ceramide protects hippocampal neurons against excitotoxic and oxidative insults, and amyloid beta-peptide toxicity. J. Neurochem. 6, 869–872.Google Scholar
  20. Guo Q., Sebastian L., Sopher B. L., Miller M. W., Glazner G. W., Ware C. B., et al. (1999) Neurotrophic factors [activity-dependent neurotrophic factor (ADNF) and basic fibroblast growth factor (bFGF)] interrupt excitotoxic neurodegenerative cascades promoted by a PS1 mutation. Proc. Natl. Acad. Sci. USA 96, 4125–4130.PubMedCrossRefGoogle Scholar
  21. Hannun Y. A. and Obeid L. M. (1997) Ceramide and the eukaryotic stress response. Biochem. Soc. Trans. 25, 1171–1175.PubMedGoogle Scholar
  22. Hara H., Fink K., Endres M., Friedlander R. M., Gagliardini V., Yuan J., and Moskowitz M. A. (1997) Attenuation of transient focal cerebral ischemic injury in transgenic mice expressing a mutant ICE inhibitory protein. J. Cereb. Blood Flow Metab. 17, 370–375.PubMedCrossRefGoogle Scholar
  23. Hartfield P. J., Mayne G. C., and Murray A. W. (1997) Ceramide induces apoptosis in PC12 cells. FEBS Lett. 401, 148–152.PubMedCrossRefGoogle Scholar
  24. Herr I., Martin-Villalba A., Kurz E., Roncaioli P., Schenkel J., Cifone M. G., and Debatin K. M. (1999) FK506 prevents stroke-induced generation of ceramide and apoptosis signaling. Brain Res. 826, 210–219.PubMedCrossRefGoogle Scholar
  25. Hida H., Takeda M., and Soliven B. (1998) Ceramide inhibits inwardly rectifying K+ currents via a Ras- and Raf-1-dependent pathway in cultured oligodendrocytes. J. Neurosci. 18, 8712–8719.PubMedGoogle Scholar
  26. Hofmann K. and Dixit V. M. (1998) Ceramide in apoptosis: does it really matter? Trends Biochem. Sci. 23, 374–377.PubMedCrossRefGoogle Scholar
  27. Horinouchi K., Erlich S., Perl D. P., Ferlinz K., Bisgaier C. L., Sandhoff K., et al. (1995) Acid sphingomyelinase deficient mice: a model of types A and B Niemann-Pick disease. Nat. Gen. 10, 288–293.CrossRefGoogle Scholar
  28. Jarvis W. D., Kolesnick R. N., Fornari F. A., Traylor R. S., Gewirtz D. A., and Grant S. (1994) Induction of apoptotic DNA damage and cell death by activation of the sphingomyelin pathway. Proc. Natl. Acad. Sci. USA 91, 73–77.PubMedCrossRefGoogle Scholar
  29. Keller J. N., Kindy M. S., Holtsberg F. W., St Clair D. K., Yen H. C., Germeyer A., et al. (1998) Mitochondrial manganese superoxide dismutase prevents neural apoptosis and reduces ischemic brain injury: suppression of peroxynitrite production, lipid peroxidation, and mitochondrial dysfunction. J. Neurosci. 18, 687–697.PubMedGoogle Scholar
  30. Kobrinsky E., Spielman A. I., Rosenzweig S., and Marks A. R. (1999) Ceramide triggers intracellular calcium release via the IP(3) receptor in Xenopus laevis oocytes. Am. J. Physiol. 277, C665–672.PubMedGoogle Scholar
  31. Kubota M., Kitahara S., Shimasaki H., and Ueta N. (1989) Accumulation of ceramide in ischemic human brain of an acute case of cerebral occlusion. Jpn. J. Exp. Med. 59, 59–64.PubMedGoogle Scholar
  32. Loddick S. A., Turnbull A. V., and Rothwell N. J. (1998) Cerebral interleukin-6 is neuroprotective during permanent focal cerebral ischemia in the rat. J. Cereb. Blood Flow Metab. 18, 176–179.PubMedCrossRefGoogle Scholar
  33. Long S. D. and Pekala P. H. (1996) Lipid mediators of insulin resistance: ceramide signalling down-regulates GLUT4 gene transcription in 3T3-L1 adipocytes. Biochem. J. 319, 179–184.PubMedGoogle Scholar
  34. Love S. (1998) Oxidative stress in brain ischemia. Brain Pathol. 9, 119–131.CrossRefGoogle Scholar
  35. Mansat-de Mas V., Bezombes C., Quillet-Mary A., Bettaieb A., D’orgeix A. D., Laurent G., and Jaffrezou J. P. (1999) Implication of radical oxygen species in ceramide generation, c-Jun N-terminal kinase activation and apoptosis induced by daunorubicin. Mol. Pharmacol. 56, 867–874.PubMedGoogle Scholar
  36. Mathias S., Pena L. A., and Kolesnick R. N. (1998) Signal transduction of stress via ceramide. Biochem. J. 335, 465–480.PubMedGoogle Scholar
  37. Mattson M. P. (1997) Neuroprotective signal transduction: relevance to stroke. Neurosci. Biobehav. Rev. 21, 193–206.PubMedCrossRefGoogle Scholar
  38. Mattson M. P., Goodman Y., Luo H., Fu W., and Furukawa K. (1997) Activation of NF-κB protects hippocampal neurons against oxidative stress-induced apoptosis: evidence for induction of manganese superoxide dismutase and suppression of peroxynitrite production and protein tyrosine nitration. J. Neurosci. Res. 49, 681–697.PubMedCrossRefGoogle Scholar
  39. Merrill, A. H. Jr., Wang E., Mullins R. E., Jamison W. C., Nimkar S., and Liotta D. (1988) Quantitation of free sphingosine in liver by high-performance liquid chromatography. Anal. Biochem. 171, 373–381.PubMedCrossRefGoogle Scholar
  40. Nikolova-Karakashian M. N., Morgan E. T., Alexander C., Liotta D. C., and Merrill A. H. (1997) Biomdal regulation of ceramidase by interleukin-1β: Implication for the regulation of cytochrome P450 2C11 (CYP2C11). J. Biol. Chem. 272, 18,718–18,724.CrossRefGoogle Scholar
  41. Pruschy M., Resch H., Shi Y. Q., Aalame N., Glanzmann C., and Bodis S. (1999) Ceramide triggers p53-dependent apoptosis in genetically defined fibrosarcoma tumour cells. Br. J. Cancer 80, 693–698.PubMedCrossRefGoogle Scholar
  42. Relton J. K. and Rothwell N. J. (1992) Interleukin-1 receptor antagonist inhibits ischaemic and excitotoxic neuronal damage in the rat. Brain Res. Bull. 29, 243–246.PubMedCrossRefGoogle Scholar
  43. Santana P., Pena L. A., Haimovitz-Friedman A., Martin S., Green D., McLoughlin M., et al. (1996) Acid sphingomyelinase-deficient human lymphoblasts and mice are defective in radiation-induced apoptosis. Cell 86, 189–199.PubMedCrossRefGoogle Scholar
  44. Scheid M. P., Foltz I. N., Young P. R., Schrader J. W., and Duronio V. (1999) Ceramide and cyclic adenosine monophosphate (cAMP) induce cAMP response element binding protein phosphorylation via distinct signaling pathways while having opposite effects on myeloid cell survival. Blood 93, 217–225.PubMedGoogle Scholar
  45. Schielke G. P., Yang G. Y., Shivers B. D., and Betz A. L. (1998) Reduced ischemic brain injury in interleukin-1β converting enzyme-deficient mice. J. Cereb. Blood Flow Metab. 18, 180–185.PubMedCrossRefGoogle Scholar
  46. Schutze S., Potthoff K., Machleidt T., Berkovic D., Wiegmann K., and Kronke M. (1992) TNF activates NF-κB by phosphatidylcholine-specific phospholipase C-induced acidic sphingomyelin breakdown. Cell 71, 765–776.PubMedCrossRefGoogle Scholar
  47. Shioda S., Ozawa H., Dohi K., Mizushima H., Matsumoto K., Nakajo S., et al. (1998) PACAP protects hippocampal neurons against apoptosis: involvement of JNK/SAPK signaling pathway. Ann. N.Y. Acad. Sci. 865, 111–117.PubMedCrossRefGoogle Scholar
  48. Wang Y. M., Seibenhener M. L., Vandenplas M. L., and Wooten M. W. (1999) Atypical PKC zeta is activated by ceramide, resulting in coactivation of NF-κB/JNK kinase and cell survival. J. Neurosci. Res. 55, 293–302.PubMedCrossRefGoogle Scholar
  49. Yoshimura S., Banno Y., Nakashima S., Hayashi K., Yamakawa H., Sawada M., et al. (1999) Inhibition of neutral sphingomyelinase activation and ceramide formation by glutathione in hypoxic PC12 cell death. J. Neurochem. 73, 675–683.PubMedCrossRefGoogle Scholar
  50. Yu Z., Zhou D., Bruce-Keller A. J., Kindy M. S., and Mattson M. P. (1999) Lack of the p50 subunit of nuclear factor-κB increases the vulnerability of hippocampal neurons to excitotoxic injury. J. Neurosci. 19, 8856–8865.PubMedGoogle Scholar

Copyright information

© Humana Press Inc 2001

Authors and Affiliations

  • Zai Fang Yu
    • 1
  • Mariana Nikolova-Karakashian
    • 2
  • Daohong Zhou
    • 3
  • Guanjun Cheng
    • 1
  • Edward H. Schuchman
    • 4
  • Mark P. Mattson
    • 1
    • 5
  1. 1.Sanders-Brown Research Center on AgingUniversity of KentuckyLexington
  2. 2.Department of PhysiologyUniversity of KentuckyLexington
  3. 3.Department of MedicineUniversity of KentuckyLexington
  4. 4.Department of Human GeneticsMount Sinai School of MedicineNew York
  5. 5.Laboratory of NeurosciencesNational Institute on AgingBaltimore

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