Novel dimeric acetylcholinesterase inhibitor bis(7)-tacrine, but not donepezil, prevents glutamate-induced neuronal apoptosis by blocking N-methyl-D-aspartate receptors

Wenming Li, Rongbiao Pi, Hugh H.N. Chan, Hongjun Fu, Nelson T.K. Lee, Hing Wai Tsang, Yongmei Pu, Donald C. Chang, Chaoying Li, Jialie Luo, Keming Xiong, Zhiwang Li, Hong Xue, Paul R. Carlier, Yuanping Pang, Karl W.K. Tsim, Mingtao Li, Yifan Han

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Abstract

The neuroprotective properties of bis(7)-tacrine, a novel dimeric acetylcholinesterase (AChE) inhibitor, on glutamate-induced excitotoxicity were investigated in primary cultured cerebellar granule neurons (CGNs). Exposure of CGNs to 75 μM glutamate resulted in neuronal apoptosis as demonstrated by Hoechst staining, TUNEL, and DNA fragmentation assays. The bis(7)-tacrine treatment (0.01-1 μM) on CGNs markedly reduced glutamate-induced apoptosis in dose- and time-dependent manners. However, donepezil and other AChE inhibitors, even at concentrations of inhibiting AChE to the similar extents as 1 μM bis(7)-tacrine, failed to prevent glutamate-induced excitotoxicity in CGNs; moreover, both atropine and dihydro-β-erythroidine, the cholinoreceptor antagonists, did not affect the anti-apoptotic properties of bis(7)-tacrine, suggesting that the neuroprotection of bis(7)-tacrine appears to be independent of inhibiting AChE and cholinergic transmission. In addition, ERK1/2 and p38 pathways, downstream signals of N-methyl-D-aspartate (NMDA) receptors, were rapidly activated after the exposure of glutamate to CGNs. Bis(7)-tacrine inhibited the apoptosis and the activation of these two signals with the same efficacy as the co-application of PD98059 and SB203580. Furthermore, using fluorescence Ca2+ imaging, patch clamp, and receptor-ligand binding techniques, bis(7)-tacrine was found effectively to buffer the intracellular Ca2+ increase triggered by glutamate, to reduce NMDA-activated currents and to compete with [3H]MK-801 with an IC50 value of 0.763 μM in rat cerebellar cortex membranes. These findings strongly suggest that bis(7)-tacrine prevents glutamate-induced neuronal apoptosis through directly blocking NMDA receptors at the MK-801-binding site, which offers a new and clinically significant modality as to how the agent exerts neuroprotective effects.

Original languageEnglish (US)
Pages (from-to)18179-18188
Number of pages10
JournalJournal of Biological Chemistry
Volume280
Issue number18
DOIs
StatePublished - May 6 2005

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Li, W., Pi, R., Chan, H. H. N., Fu, H., Lee, N. T. K., Tsang, H. W., Pu, Y., Chang, D. C., Li, C., Luo, J., Xiong, K., Li, Z., Xue, H., Carlier, P. R., Pang, Y., Tsim, K. W. K., Li, M., & Han, Y. (2005). Novel dimeric acetylcholinesterase inhibitor bis(7)-tacrine, but not donepezil, prevents glutamate-induced neuronal apoptosis by blocking N-methyl-D-aspartate receptors. Journal of Biological Chemistry, 280(18), 18179-18188. https://doi.org/10.1074/jbc.M411085200