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Strategies for Study of Neuroprotection from Cold-preconditioning
Published on: September 2, 2010
Preconditioning neuroprotection in global cerebral ischemia involves NMDA receptor-mediated ERK-JNK3 crosstalk
Quan-Guang Zhang1, Rui-Min Wang, Dong Han
1Developmental Neurobiology Program, Institute of Molecular Medicine and Genetics, and Department of Neurology, Medical College of Georgia, Augusta, GA 30912, USA.
Abstract:
Previous work has demonstrated that ischemic preconditioning neuroprotection is associated with inhibition of JNK pathway activation. The present study was designed to examine the hypothesis that the suppression of JNK3 activation by preconditioning is mediated by NMDA receptors and crosstalk between ERK1/2 and JNK3. Preconditioning (3 min ischemia) 2 days before global cerebral ischemia (8-min) markedly decreased neuronal degeneration in hippocampus CA1, an effect abolished by pretreatment with the NMDA receptor antagonist, MK-801. Furthermore, preconditioning abolished cerebral ischemia-induced JNK3 activation and enhanced ERK1/2 activation, an effect reversed by MK-801. Due to the inverse relationship between ERK1/2 and JNK3 activation following preconditioning, we hypothesized that ERK1/2 may regulate JNK3 activation following preconditioning. In support of this contention, pretreatment with the MEK inhibitor, PD98059 significantly attenuated preconditioning-induced ERK1/2 phosphorylation, and strongly reversed preconditioning down-regulation of JNK3 phosphorylation. This finding suggests that ERK1/2 signaling is responsible for preconditioning-induced down-regulation of JNK3 activation. Western blot analysis and immunohistochemistry further demonstrated that preconditioning, in an NMDA-dependent manner, enhanced activation of the pro-survival factors, p-CREB and Bcl-2, while attenuating activation of putative pro-death factors, p-c-Jun and Fas-L in the hippocampus CA1. As a whole, the study demonstrates that preconditioning attenuation of pro-death JNK3 in the hippocampus CA1 following global cerebral ischemia is mediated by NMDA receptor-induced crosstalk between ERK1/2 and JNK3. The ERK1/2-mediated reduction of JNK3 activation leads to enhanced pro-survival signaling (P-CREB and Bcl-2 induction) and attenuation of pro-death signaling (p-c-Jun and Fas-L), with subsequent induction of ischemic tolerance.
Insights
Ischemic preconditioning protects neurons by inhibiting JNK3 activation via NMDA receptors and ERK1/2 signaling. This pathway enhances pro-survival factors and reduces pro-death factors, inducing tolerance to cerebral ischemia.
Area of Science:
- Neuroscience
- Molecular Biology
- Cell Signaling
Background:
- Ischemic preconditioning confers neuroprotection against cerebral ischemia.
- This protection is linked to the inhibition of c-Jun N-terminal kinase (JNK) pathway activation.
- The precise molecular mechanisms, particularly the roles of NMDA receptors and extracellular signal-regulated kinases (ERK1/2), remain to be fully elucidated.
Purpose of the Study:
- To investigate the hypothesis that NMDA receptor activation and crosstalk between ERK1/2 and JNK3 mediate the suppression of JNK3 activation by ischemic preconditioning.
- To elucidate the downstream signaling events, including pro-survival and pro-death factors, influenced by this preconditioning-induced pathway.
Main Methods:
- Global cerebral ischemia model in rodents with preconditioning (3 min ischemia) applied 2 days prior to the main ischemic insult (8 min).
- Pharmacological interventions using NMDA receptor antagonist (MK-801) and MEK inhibitor (PD98059).
- Western blot analysis and immunohistochemistry to assess the activation of JNK3, ERK1/2, p-CREB, Bcl-2, p-c-Jun, and Fas-L in the hippocampus CA1 region.
Main Results:
- Preconditioning significantly reduced neuronal degeneration in the hippocampus CA1.
- This neuroprotection was abolished by MK-801, indicating NMDA receptor dependence.
- Preconditioning inhibited ischemia-induced JNK3 activation and enhanced ERK1/2 activation, effects reversed by MK-801.
- MEK inhibition (PD98059) attenuated ERK1/2 phosphorylation and reversed the down-regulation of JNK3 phosphorylation, suggesting ERK1/2 mediates JNK3 suppression.
- Preconditioning, in an NMDA-dependent manner, enhanced pro-survival factors (p-CREB, Bcl-2) and attenuated pro-death factors (p-c-Jun, Fas-L).
Conclusions:
- Ischemic preconditioning attenuates JNK3 activation in the hippocampus CA1 following global cerebral ischemia through an NMDA receptor-dependent mechanism involving crosstalk with ERK1/2 signaling.
- The ERK1/2-mediated reduction in JNK3 activation promotes pro-survival signaling and suppresses pro-death signaling, ultimately inducing ischemic tolerance.
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