NMDA receptor dependent and independent immediate early gene expression induced by focal mechanical brain injury.

In the present study we analysed, by in situ hybridization, the effects of an extremely localized mechanical brain injury, obtained by the simple needle insertion (30 g) in rat hippocampus or cortex, on the expression of several immediate early genes (c-fos, fosB, c-jun, junB, junD, zif/268). When the needle is deepened into the hippocampus through the cortex, a simultaneous ipsilateral activation of all examined IEGs is observed in both the cerebral cortex and in the dentate gyrus of hippocampus. Maximal effects are detected between 30 and 60 min with the following rank order of induction: zif/268 > c-fos- > junB > fosB > c-jun > junD. On the other hand, when the penetration of the needle is limited to the cerebral cortex the activation of the IEGs (c-fos, fosB, junB and zif/268) spreads throughout the ipsilateral cortex but does not involve the hippocampal region. Systemic administration of ketamine, a non-competitive antagonist of N-methyl-D-aspartate (NMDA) receptors, blocks IEG expression induced by brain injury in the cerebral cortex and in the hippocampal dentate gyrus. Pretreatment with the anticonvulsant diazepam, the anaesthetic urethane, or the muscarinic receptor antagonist scopolamine do not affect the injury-induced genomic response. An important regional difference in the sensitivity to the blocking effect of ketamine can be observed analysing the results regarding the zif/268 gene expression in the hippocampus. A clear induction of this gene by needle insertion can be detected both in the dentate gyrus and in the hippocampal layers. However, the dentate gyrus induction is completely blocked by the ketamine pretreatment, while the induction in the hippocampal layers is not affected by this NMDA antagonist. The zif/268 induction in the hippocampal layers is not blocked even if the intracerebroventricular administration of a non-NMDA glutamate receptor antagonist is associated to the systemic pretreatment with ketamine. This result represents the first observation of injury-induced neuronal genomic responses that are not critically dependent on the NMDA receptor activity.