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C-Jun N-terminal kinases/c-Jun and p38 pathways cooperate in ceramide-induced neuronal apoptosis

C-Jun N-terminal kinases/c-Jun and p38 pathways cooperate in ceramide-induced neuronal apoptosis
C-Jun N-terminal kinases/c-Jun and p38 pathways cooperate in ceramide-induced neuronal apoptosis
Understanding the regulation of the apoptotic program in neurons by intracellular pathways is currently a subject of great interest. Recent results suggest that c-Jun N-terminal kinases (JNK), mitogen-activated protein kinases and the transcription factor c-Jun are important regulators of this cell death program in post-mitotic neurons following survival-factor withdrawal. Our study demonstrates that ceramide levels increase upon survival-factor withdrawal in primary cultured cortical neurons. Furthermore, survival-factor withdrawal or addition of exogenous c2-ceramide induces JNK pathway activation in these cells. Western blot analyses of JNK and c-Jun using phospho-specific antibodies reveal that JNK and subsequent c-Jun phosphorylation occur hours before the initiation of apoptosis, reflected morphologically by neurite retraction and fragmentation, cell-body shrinkage and chromatin fragmentation. Immunocytochemistry using the same antibodies shows that phospho-JNK are localized in the neurites of control neurons and translocate to the nucleus where phospho-c-Jun concurrently appears upon ceramide-induced apoptosis. To determine if ceramide-induced c-Jun activation is responsible for the induction of the apoptotic program, we performed transient transfections of a dominant negative form of c-Jun, truncated in its transactivation region. Our results show that DNc-Jun partially protects cortical neurons from ceramide-induced apoptosis. Treatment of dominant negative c-Jun-expressing neurons with the pharmacological inhibitor of p38 kinase, SB203580, completely blocked neuronal death. Thus our data show that p38 and JNK/c-Jun pathways cooperate to induce neuronal apoptosis.
primary cultures, SAPK, serum withdrawal, neuronaldeath
0306-4522
387-397
Willaime-Morawek, S.
24a2981f-aa9e-4bf6-ad12-2ccf6b49f1c0
Brami-Cherrier, K.
1d70f208-a693-4595-86cb-81eb4f2ba7d3
Mariani, J.
d41a573a-a1ac-4c1b-b8d3-d97fad4756d1
Caboche, J.
071c14c4-71c6-435d-9725-8bde4e2f6447
Brugg, B.
47c3b9cc-b1ab-4a9c-addd-a100221e513a
Willaime-Morawek, S.
24a2981f-aa9e-4bf6-ad12-2ccf6b49f1c0
Brami-Cherrier, K.
1d70f208-a693-4595-86cb-81eb4f2ba7d3
Mariani, J.
d41a573a-a1ac-4c1b-b8d3-d97fad4756d1
Caboche, J.
071c14c4-71c6-435d-9725-8bde4e2f6447
Brugg, B.
47c3b9cc-b1ab-4a9c-addd-a100221e513a

Willaime-Morawek, S., Brami-Cherrier, K., Mariani, J., Caboche, J. and Brugg, B. (2003) C-Jun N-terminal kinases/c-Jun and p38 pathways cooperate in ceramide-induced neuronal apoptosis. Neuroscience, 119 (2), 387-397. (doi:10.1016/S0306-4522(02)00996-X).

Record type: Article

Abstract

Understanding the regulation of the apoptotic program in neurons by intracellular pathways is currently a subject of great interest. Recent results suggest that c-Jun N-terminal kinases (JNK), mitogen-activated protein kinases and the transcription factor c-Jun are important regulators of this cell death program in post-mitotic neurons following survival-factor withdrawal. Our study demonstrates that ceramide levels increase upon survival-factor withdrawal in primary cultured cortical neurons. Furthermore, survival-factor withdrawal or addition of exogenous c2-ceramide induces JNK pathway activation in these cells. Western blot analyses of JNK and c-Jun using phospho-specific antibodies reveal that JNK and subsequent c-Jun phosphorylation occur hours before the initiation of apoptosis, reflected morphologically by neurite retraction and fragmentation, cell-body shrinkage and chromatin fragmentation. Immunocytochemistry using the same antibodies shows that phospho-JNK are localized in the neurites of control neurons and translocate to the nucleus where phospho-c-Jun concurrently appears upon ceramide-induced apoptosis. To determine if ceramide-induced c-Jun activation is responsible for the induction of the apoptotic program, we performed transient transfections of a dominant negative form of c-Jun, truncated in its transactivation region. Our results show that DNc-Jun partially protects cortical neurons from ceramide-induced apoptosis. Treatment of dominant negative c-Jun-expressing neurons with the pharmacological inhibitor of p38 kinase, SB203580, completely blocked neuronal death. Thus our data show that p38 and JNK/c-Jun pathways cooperate to induce neuronal apoptosis.

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Published date: 27 June 2003
Keywords: primary cultures, SAPK, serum withdrawal, neuronaldeath

Identifiers

Local EPrints ID: 66392
URI: http://eprints.soton.ac.uk/id/eprint/66392
ISSN: 0306-4522
PURE UUID: f63c24b4-3006-444f-a645-097b854805da
ORCID for S. Willaime-Morawek: ORCID iD orcid.org/0000-0002-1121-6419

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Date deposited: 10 Jun 2009
Last modified: 14 Mar 2024 02:53

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Contributors

Author: K. Brami-Cherrier
Author: J. Mariani
Author: J. Caboche
Author: B. Brugg

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