Abstract
Cytidine 5′-diphosphocholine (CDP-choline) has been shown to reduce neuronal degeneration induced in central nervous system (CNS) injury. However, the precise mechanism underlying the neuroprotective properties of this molecule is still unknown. Excitotoxicity causes cell death in CNS injury (trauma or ischemia) and has also been involved in neurodegenerative diseases. We have examined whether CDP-choline prevents glutamate-mediated cell death, determined by trypan blue exclusion and lactate dehydrogenase activity assays. Pretreatment of rat cerebellar granule cells (CGCs) with CDP-choline causes a dose- and time-dependent reduction of glutamate-induced excitotoxicity. Cell death is prevented >50% when 100 µM CDP-choline is added 6 d before the glutamate excitotoxic insult but less than 20% when added concomitantly with glutamate. Pretreatment of CGCs with CDP-choline reduces almost completely (>80%) the number of apoptotic cells analyzed by flow cytometry, suggesting that CDP-choline exerts a neuroprotective effect by inhibiting the apoptotic pathway induced by glutamate.
Similar content being viewed by others
References
Alvarez A. A., Sampedro C., Lozano R., and Cacabelos R. (1999) Citicoline protects hippocampal neurons against apoptosis induced by brain β-amyloid deposits plus cerebral hypoperfusion in rats. Methods Find. Exp. Clin. Pharmacol. 21(8), 535–540.
Ankarcrona M., Dypbukt J., Bonfoco E., Zhivotovsky B., Orrenius S., Lipton S. A., and Nicotera P. (1995) Glutamate-induced neuronal death: a succession of necrosis or apoptosis depending on mitochondrial function. Neuron 15, 961–973.
Ankarcrona M., Dypbukt J., Orrenius S., and Nicotera P. (1996) Calcineurin and mitochondrial function in glutamate-induced neuronal cell death. FEBS Lett. 394, 321–324.
Ariga T., Jarvis W. D., and Yu R. K. (1998) Role of sphingolipid-mediated cell death in neurodegenerative diseases. J. Lipid Res. 39, 1–16.
Bachis A., Colangelo A. M., Vicini S., Doe P. P., De Bernardi M. A., Brooker G., and Mocchetti I. (2001) Interleukin-10 prevents glutamate-mediated cerebellar granule cell death by blocking caspase-3-like activity. J. Neurosci. 21(9), 3104–3112.
Baskaya M. K., Dogan A., and Dempsey, R. J. (2000) Neuroprotective effects of citicoline on brain edema and blood-brain barrier breakdown after traumatic brain injury. J. Neurosury. 92, 448–452.
Blusztajn J. K. (1998) Choline, a vital amine. Science 281, 794–795.
Brandoli C., Sanna A., De Bernardi M. A., Follesa P., Brooker G., Mocchetti I. (1998) Brain derived neurotrophic factor and basic fibroblast growth factor downregulate NMDA receptor function in cerebellar granule cells. J. Neurosci. 18, 7953–7961.
Clark W. M., Wechsler L. R., Sabounjian L. A., and Schiderski U. E. (2001) A phase III randomized efficacy trial of 2000 mg citicoline in acute ischemic stroke patients. Neurology 57(9), 1595–1602.
Dixon C. E., Ma X. C., and Marion D. W. (1997) Effects of CDP-choline treatment on neurobehavioral deficits after TBI and on hippocampal and neocortical acetylcholine release. J. Neurotrauma 14, 161–169.
Gill J. S. and Windebank A. J. (2000) Ceramide initiates Nf-κβ-mediated caspase activation in neuronal apoptosis. Neurobiol. Dis. 7, 448–461.
Kaasik A., Kalda A., Jaako K., and Zharkovsky A. (2001) Dehydroepiandrosterone sulfate prevents oxygen-glucose deprivation-induced injury in cerebellar granule cell culture. Neuroscience 102, 427–432.
Miguel-Hidalgo J. J., Alvarez X. A., Lagares R., Franco A., Fernandez-Novoa L., and Cacabelos R. (1996) Brain neurotoxic lesions in rats: study of the neuroprotective effects of citicoline. Eur. Neuropsychopharmacol. 6 (Suppl. 3), 193–194.
Nonaka S., Hough C. J., and Chuang D. M. (1998) Chronic lithium treatment robustly protects neurons in the central nervous system against excitotoxicity by inhibiting N-methyl-D-aspartate receptor mediated calcium influx. Proc. Natl. Acad. Sci. USA. 95, 2642–2647.
Olmos G., DeGregorio-Rocasolano N., Regalado M. P., Gasull T., Boronat M. A., Trullas R., et al. (1999) Protection by imidazol(ine) drugs and agmantine of glutamate-induced neurotoxicity in cultured cerebellar granule cells through blockade of NMDA receptor. Br. J. Pharmacol. 127(6), 1317–1326.
Pelled D., Raveh T., Riebeling C., Fridkin M., Berissi H., Futerman A. H., and Kimchi A. (2002) Death associated protein (DAP) kinase plays a central role in ceramide-induced apoptosis in cultured hippocampal neurons. J. Biol. Chem. 277(3), 1957–1961.
Ramos B., Salido G. M., Campo M. L., and Claro E. (2000) Inhibition of phosphatidylcholine precedes apoptosis induced by C2-ceramide: protection by exogenous phosphatidylcholine. Neuroreport 11, 3103–3108.
Rao A. M., Hatcher J. F., and Dempsey R. J. (1999) DCP-choline: neuroprotection in transient forebrain ischemia of gerbils J. Neurosci. Res. 58, 697–705.
Rao A. M., Hatcher J. F., and Dempsey R. J. (2000) Lipid alterations in transient forebrain ischemia: possible new mechanisms of CDP-choline neuroprotection. J. Neurochem. 75, 2528–2535.
Shuaib A., Yang Y., Li Q. (2000) Evaluating the efficacy of citicoline in embolic ischemic stroke in rats: neuroprotective effects when used alone or in combination with urokinase. Exp. Neurol. 161, 733–739.
Uberti D., Belloni M., Grilli M., Spano P. F., and Memo M. (1998) Induction of tumor-supressor phosphoprotein p53 in the apoptosis of cultured rat cerebellar neurones triggered by excitatory amino acids. Eur. J. Neurosci. 10, 246–254.
Uberti D., Grilli M., and Memo M. (2000) Contribution of NF-κβ and p53 in the glutamate-induced apoptosis. Int. J. Dev. Neurosci. 18, 447–454.
Willaime S., Vanhoutte P., Caboche J., Lemaigre-Dubreuil Y., Mariani J., and Brugg B. (2001) Ceramide-induced apoptosis in cortical neurons is mediated by an increase in p38 phosphorylation and not by the decrease in ERK phosphorylation. Eur. J. Neurosci. 13, 2037–2046.
Yen C. L., Mar M. H., Meeker R. B., Fernandes A., and Zeisel S. H. (2001) Choline deficiency induces apoptosis in primary cultures of fetal neurons. FASEB J. 15(10), 1704–1710.
Yen C.-L. E., Mar M.-H., and Zeisel, S. H. (1999) Choline deficiency-induced apoptosis in PC12 cells is associated with diminished membrane phosphatidylcholine and sphingomyelin, accumulation of ceramide and diacylglycerol, and activation of a caspase. FASEB J. 13, 135–142.
Yu Z. F., Nikolova-Karakashian M., Zhou D., Cheng G., Schuchuman E. H., and Mattson M. P. (2000) Pivotal role for acidic sphingomyelinase in cerebral ischemia-induced ceramide and cytokine production, and neuronal apoptosis. J. Mol. Neurosci. 15(2), 85–97.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Mir, C., Clotet, J., Aledo, R. et al. CDP-choline prevents glutamate-mediated cell death in cerebellar granule neurons. J Mol Neurosci 20, 53–59 (2003). https://doi.org/10.1385/JMN:20:1:53
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1385/JMN:20:1:53