Alternative Splicing at the C‐terminal but not at the N‐terminal Domain of the NMDA Receptor NR1 is Altered in the Kindled Hippocampus

Abstract

Several lines of evidence suggest that N-methyl-D-aspartate (NMDA) receptors significantly contribute to the development of kindling. In addition, a lasting enhancement of the NMDA receptor function has been suggested to play a significant role in the chronic hyperexcitability occurring in the hippocampus after kindling epileptogenesis. We have investigated whether hippocampal kindling induces changes in the NMDA receptor at the molecular level by assessing the expression of mRNAs of the different spliced variants at the N-terminal (exon 5) and C-terminal (exon 21) position of the NMDA receptor 1 (NR1) gene by means of the reverse transcription-polymerase chain reaction. Alternative splicing at exon 5 confers different sensitivity of the NMDA receptor to polyamines while exon 21 encodes a 37-amino acid insert containing the major phosphorylation sites for protein kinase C. One week after the acquisition of stage 5 of kindling in rats (generalized tonic-clonic seizures), the relative abundance of the two alternatively spliced forms at the C-terminal domain, respectively containing (+) or lacking (-) exon 21, was reversed compared to controls (implanted with electrodes but not stimulated) in the dorsal hippocampus ipsilateral and contralateral to the electrical stimulation. The exon 21+/exon 21- mRNA ratio for controls was 1.3 +/- 0.04 (mean +/- SE); for ipsilaterally kindled rats it was 0.64 +/- 0.05 (P < 0.05), and for contralaterally kindled rats it was 0.48 +/- 0.07 (P < 0.01). Similar bilateral effects were observed in the ventral hippocampus (temporal pole). No changes were found 4 weeks after stage 5 seizures and 1 week after the induction of a single afterdischarge. No significant alterations were induced by kindling in the relative abundance of the spliced variants containing or lacking exon 5. Our findings show selective changes in alternative splicing of the NR1 gene after repeated application of an epileptogenic stimulus. This may generate receptors with different functional properties, which may contribute to the increased sensitivity for the induction of generalized seizures during kindling.

Activity‐dependent regulation of alternative splicing patterns in the rat brain

“…For example, we detected a modest response of NMDAR1 receptor exon 21 after pilocarpine treatment only in cortex. A similar observation was made by Vezzani et al . (1995); prolonged kindling of rats was shown to change this splice variant after 1 week.…”

Development of Post-Traumatic Epilepsy after Controlled Cortical Impact and Lateral Fluid-Percussion-Induced Brain Injury in the Mouse

“…In models in which epileptogenesis has been triggered by an acquired insult, that is, status epilepticus (SE) or TBI, the scant literature available suggests that perhaps it can. Rats with spontaneous seizures triggered by kainic acid or pilocarpineinduced SE at 30-45 days before a PTZ test showed enhanced susceptibility to PTZ-induced generalized convulsions and SE (Vezzani et al, 1995;Blanco et al, 2009). Further, the latency to the first generalized convulsion was reduced from 1280 sec (controls) to 389 sec (post-SE animals; Blanco et al, 2009).…”

Alternative Pre-mRNA Splicing and Neuronal Function

“…There are also a number of interesting reports of the splicing of these NMDA exons being altered by cell excitation and other stimuli (Musshoff et al, 2000;Rafiki et al, 1998;Vallano et al, 1999;Vallano et al, 1996;Vezzani et al, 1995;Xie and Black, 2001). Exons 5 and 21 show divergent patterns of inclusion within the brain.…”