Abstract
Cells preferentially expressing GluR4-containing α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) receptors are particularly sensitive to excitotoxicity mediated through non-N-methyl-D-aspartate receptors. However, the excitotoxic signalling pathways associated with GluR4-containing AMPA receptors are not known. In this work, we investigated the downstream signals coupled to excitotoxicity mediated by Ca2+-permeable GluR4-containing AMPA receptors, using a HEK 293 cell line constitutively expressing the GluR4flip subunit of AMPA receptors (HEK-GluR4). Glutamate stimulation of GluR4-containing AMPA receptors decreased cell viability, in a calcium-dependent manner, when the receptor desensitisation was prevented with cyclothiazide. The excitotoxic stimulation mediated through GluR4-containing AMPA receptors increased activator protein-1 (AP-1) DNA-binding activity. Inhibition of the AP-1 activity by overexpression of a c-Jun dominant-negative form protected HEK-GluR4 cells against excitotoxic damage. Taken together, the results indicate that overactivation of Ca2+-permeable GluR4-containing AMPA receptors is coupled to a death pathway mediated, at least in part, by the AP-1 transcription factor.
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Abbreviations
- HEK:
-
human embrionic kidney
- AMPA:
-
α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid
- KA:
-
kainate
- NMDA:
-
N-methyl-D-aspartate
- GluR:
-
glutamate receptor
- CNQX:
-
6-cyano-7-nitroquinoxaline-2,3-dione
- CTZ:
-
cyclothiazide
- MTT:
-
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl Tetrazolium bromide
- PBS:
-
phosphate buffered saline
- AP-1:
-
activator protein-1
- dn c-Jun:
-
c-Jun dominant negative
- JNK:
-
c-Jun N-terminal kinase
- MLK:
-
mixed lineage kinase
- ATF2:
-
activating transcription factor 2
- PSD:
-
postsynaptic density
- ALS:
-
amyotrophic lateral sclerosis
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Acknowledgements
We thank Professor Caetana M Carvalho for the numerous suggestions and for the critical reading of the manuscript. This work was supported by the Portuguese Science Foundation (FCT) FEDER, and by the Bissaya Barreto Foundation.
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Santos, A., Duarte, C., Iizuka, M. et al. Excitotoxicity mediated by Ca2+-permeable GluR4-containing AMPA receptors involves the AP-1 transcription factor. Cell Death Differ 13, 652–660 (2006). https://doi.org/10.1038/sj.cdd.4401785
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DOI: https://doi.org/10.1038/sj.cdd.4401785
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