Elsevier

Neuroscience

Volume 156, Issue 3, 15 October 2008, Pages 436-449
Neuroscience

Behavioural neuroscience
Differential expression of phosphorylated Ca2+/calmodulin-dependent protein kinase II and phosphorylated extracellular signal-regulated protein in the mouse hippocampus induced by various nociceptive stimuli

https://doi.org/10.1016/j.neuroscience.2008.08.002Get rights and content

Abstract

In the present study, we characterized differential expressions of phosphorylated Ca2+/calmodulin-dependent protein kinase IIα (pCaMKIIα) and phosphorylated extracellular signal-regulated protein (pERK) in the mouse hippocampus induced by various nociceptive stimuli. In an immunoblot study, s.c. injection of formalin and intrathecal (i.t.) injections of glutamate, tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1 β) significantly increased pCaMKIIα expression in the hippocampus, but i.p. injections of acetic acid did not. pERK1/2 expression was also increased by i.t. injection of glutamate, TNF-α, and IL-1β but not by s.c. injections of formalin or i.p. injections of acetic acid. In an immunohistochemical study, we found that increased pCaMKIIα and pERK expressions were mainly located at CA3 or the dentate gyrus of the hippocampus. In a behavioral study, we assessed the effects of PD98059 (a MEK 1/2 inhibitor) and KN-93 (a CaMKII inhibitor) following i.c.v. administration on the nociceptive behaviors induced by i.t. injections of glutamate, pro-inflammatory cytokines (TNF-α or IL-1β), and i.p. injections of acetic acid. PD98059 as well as KN-93 significantly attenuated the nociceptive behavior induced by glutamate, pro-inflammatory cytokines, and acetic acid. Our results suggest that (1) pERKα and pCaMK-II located in the hippocampus are important regulators during the nociceptive processes induced by s.c. formalin, i.t. glutamate, i.t. pro-inflammatory cytokines, and i.p. acetic acid injection, respectively, and (2) the alteration of pERK and pCaMKIIα in nociceptive processing induced by formalin, glutamate, pro-inflammatory cytokines and acetic acid was modulated in a different manner.

Section snippets

Experimental procedures

These experiments were approved by the Hallym University Animal Care and Use Committee. All procedures were conducted in accordance with the ‘Guide for Care and Use of Laboratory Animals’ published by the National Institutes of Health and the ethical guidelines of the International Association for the Study of Pain. We did our best to minimize the number of animals used and their suffering.

The effect of various nociceptive stimuli on pERK1/2 and pCaMK-IIα expression in the mouse hippocampus

All Western blot data shown in this study were from experiments repeated at least three times (the same three mice tested three times). To examine the effects of various nociceptive stimuli (1% acetic acid i.p. injection, 5% formalin s.c. injection, glutamate i.t. injection, and proinflammatory cytokines i.t. injection) on the hippocampal pCaMKIIα (53 kDa) and pERK1/2 (44 and 42 kDa) expression, Western blot analysis was performed at various time points (30 min, 1 h, and 2 h) after nociceptive

Discussion

We have tested the hypothesis that the hippocampal pCaMK-IIα and pERK1/2 are involved in nociceptive processing. We found that the expressions of pCaMK-IIα and pERK1/2 were significantly increased by various nociceptive stimuli. In addition, supraspinally administered CaMK-II inhibitor (KN-93) as well as MEK1/2 inhibitor (PD98059) significantly attenuated the nociceptive behaviors induced by i.p. acetic acids, i.t. glutamate, TNF-α, and IL-1β injection. Although the differential antinociceptive

Conclusion

In conclusion, the present data demonstrated that nociceptive stimuli induced by formalin, acetic acid, glutamate, and pro-inflammatory cytokines activate the hippocampal pCaMK-IIα and pERK1/2 molecules in a different manner, and these results suggest that alterations of hippocampal pCaMK-IIα and pERK1/2 may play an important role in supraspinal nociceptive processing.

Acknowledgments

This research was supported by research grants from the Korean Ministry of Science and Technology under the auspices of Brain Frontier (M103KV010014-08K2201-01410) and the MRC program of MOST/KOSEF (R13-2005-022-03002-0).

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