J. Cell Biol.
© The Rockefeller University Press
0021-9525/97/03/1137/13 $2.00
Volume 136, Number 5, March 10, 1997 1137-1149

Neuroprotective Action of Cycloheximide Involves Induction of Bcl-2 and Antioxidant Pathways

Katsutoshi Furukawa,^* Steven Estus,^* Weiming Fu,^* Robert J. Mark,^* and Mark P. Mattson^*§

^* Sanders-Brown Research Center on Aging,  Department of Physiology, ^§ Department of Anatomy and Neurobiology, University of Kentucky, Lexington, Kentucky 40536

The ability of the protein synthesis inhibitor cycloheximide (CHX) to prevent neuronal death in different paradigms has been interpreted to indicate that the cell death process requires synthesis of "killer" proteins. On the other hand, data indicate that neurotrophic factors protect neurons in the same death paradigms by inducing expression of neuroprotective gene products. We now provide evidence that in embryonic rat hippocampal cell cultures, CHX protects neurons against oxidative insults by a mechanism involving induction of neuroprotective gene products including the antiapoptotic gene bcl-2 and antioxidant enzymes. Neuronal survival after exposure to glutamate, FeSO₄, and amyloid -peptide was increased in cultures pretreated with CHX at concentrations of 50-500 nM; higher and lower concentrations were ineffective. Neuroprotective concentrations of CHX caused only a moderate (20-40%) reduction in overall protein synthesis, and induced an increase in c-fos, c-jun, and bcl-2 mRNAs and protein levels as determined by reverse transcription-PCR analysis and immunocytochemistry, respectively. At neuroprotective CHX concentrations, levels of c-fos heteronuclear RNA increased in parallel with c-fos mRNA, indicating that CHX acts by inducing transcription. Neuroprotective concentrations of CHX suppressed accumulation of H₂O₂ induced by FeSO₄, suggesting activation of antioxidant pathways. Treatment of cultures with an antisense oligodeoxynucleotide directed against bcl-2 mRNA decreased Bcl-2 protein levels and significantly reduced the neuroprotective action of CHX, suggesting that induction of Bcl-2 expression was mechanistically involved in the neuroprotective actions of CHX. In addition, activity levels of the antioxidant enzymes Cu/ Zn-superoxide dismutase, Mn-superoxide dismutase, and catalase were significantly increased in cultures exposed to neuroprotective levels of CHX. Our data suggest that low concentrations of CHX can promote neuron survival by inducing increased levels of gene products that function in antioxidant pathways, a neuroprotective mechanism similar to that used by neurotrophic factors.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

• Edlich, F., Weiwad, M., Wildemann, D., Jarczowski, F., Kilka, S., Moutty, M.-C., Jahreis, G., Lucke, C., Schmidt, W., Striggow, F., Fischer, G. (2006). The Specific FKBP38 Inhibitor N-(N',N'-Dimethylcarboxamidomethyl)cycloheximide Has Potent Neuroprotective and Neurotrophic Properties in Brain Ischemia. J. Biol. Chem. 281: 14961-14970 [Abstract] [Full Text]  
• Wang, J., Wei, Q., Wang, C.-Y., Hill, W. D., Hess, D. C., Dong, Z. (2004). Minocycline Up-regulates Bcl-2 and Protects against Cell Death in Mitochondria. J. Biol. Chem. 279: 19948-19954 [Abstract] [Full Text]  
• Saha, A. R., Hill, J., Utton, M. A., Asuni, A. A., Ackerley, S., Grierson, A. J., Miller, C. C., Davies, A. M., Buchman, V. L., Anderton, B. H., Hanger, D. P. (2004). Parkinson's disease {alpha}-synuclein mutations exhibit defective axonal transport in cultured neurons. J. Cell Sci. 117: 1017-1024 [Abstract] [Full Text]  
• Ryu, H., Lee, J., Olofsson, B. A., Mwidau, A., Dedeoglu, A., Escudero, M., Flemington, E., Azizkhan-Clifford, J., Ferrante, R. J., Ratan, R. R. (2003). Histone deacetylase inhibitors prevent oxidative neuronal death independent of expanded polyglutamine repeats via an Sp1-dependent pathway. Proc. Natl. Acad. Sci. USA 100: 4281-4286 [Abstract] [Full Text]  
• Spencer, M. L., Shao, H., Tucker, H. M., Andres, D. A. (2002). Nerve Growth Factor-dependent Activation of the Small GTPase Rin. J. Biol. Chem. 277: 17605-17615 [Abstract] [Full Text]  
• Torres-Roca, J. F., Tung, J. W., Greenwald, D. R., Brown, J. M., Herzenberg, L. A., Herzenberg, L. A., Katsikis, P. D. (2000). An Early Oxygen-Dependent Step Is Required for Dexamethasone-Induced Apoptosis of Immature Mouse Thymocytes. J. Immunol. 165: 4822-4830 [Abstract] [Full Text]  
• Alessandrini, A., Namura, S., Moskowitz, M. A., Bonventre, J. V. (1999). MEK1 protein kinase inhibition protects against damage resulting from focal cerebral ischemia. Proc. Natl. Acad. Sci. USA 96: 12866-12869 [Abstract] [Full Text]  
• Choi, J. W., Jung, S. E. (1999). Lovastatin-Induced Proliferation Inhibition and Apoptosis in C6 Glial Cells. J. Pharmacol. Exp. Ther. 289: 572-579 [Abstract] [Full Text]  
• Tamatani, M., Che, Y. H., Matsuzaki, H., Ogawa, S., Okado, H., Miyake, S.-i., Mizuno, T., Tohyama, M. (1999). Tumor Necrosis Factor Induces Bcl-2 and Bcl-x Expression through NFkappa B Activation in Primary Hippocampal Neurons. J. Biol. Chem. 274: 8531-8538 [Abstract] [Full Text]  
• Cunningham, T. J., Hodge, L., Speicher, D., Reim, D., Tyler-Polsz, C., Levitt, P., Eagleson, K., Kennedy, S., Wang, Y. (1998). Identification of a Survival-Promoting Peptide in Medium Conditioned by Oxidatively Stressed Cell Lines of Nervous System Origin. J. Neurosci. 18: 7047-7060 [Abstract] [Full Text]  
• Farinelli, S. E., Greene, L. A., Friedman, W. J. (1998). Neuroprotective Actions of Dipyridamole on Cultured CNS Neurons. J. Neurosci. 18: 5112-5123 [Abstract] [Full Text]  
• Chan, G. C.-K., Hinds, T. R., Impey, S., Storm, D. R. (1998). Hippocampal Neurotoxicity of Delta 9-Tetrahydrocannabinol. J. Neurosci. 18: 5322-5332 [Abstract] [Full Text]  
• Xu, J., Yeh, C.-H., Chen, S., He, L., Sensi, S. L., Canzoniero, L. M. T., Choi, D. W., Hsu, C. Y. (1998). Involvement of de Novo Ceramide Biosynthesis in Tumor Necrosis Factor-alpha /Cycloheximide-induced Cerebral Endothelial Cell Death. J. Biol. Chem. 273: 16521-16526 [Abstract] [Full Text]  
• Esch, F., Lin, K.-I, Hills, A., Zaman, K., Baraban, J. M., Chatterjee, S., Rubin, L., Ash, D. E., Ratan, R. R. (1998). Purification of a Multipotent Antideath Activity from Bovine Liver and Its Identification as Arginase: Nitric Oxide-Independent Inhibition of Neuronal Apoptosis. J. Neurosci. 18: 4083-4095 [Abstract] [Full Text]  
• Liu, X. Z., Xu, X. M., Hu, R., Du, C., Zhang, S. X., McDonald, J. W., Dong, H. X., Wu, Y. J., Fan, G. S., Jacquin, M. F., Hsu, C. Y., Choi, D. W. (1997). Neuronal and Glial Apoptosis after Traumatic Spinal Cord Injury. J. Neurosci. 17: 5395-5406 [Abstract] [Full Text]  
• Kruman, I., Bruce-Keller, A. J., Bredesen, D., Waeg, G., Mattson, M. P. (1997). Evidence that 4-Hydroxynonenal Mediates Oxidative Stress-Induced Neuronal Apoptosis. J. Neurosci. 17: 5089-5100 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents