Nuclear degradation of particular Fos family members expressed following injections of NMDA and kainate in murine hippocampus.

Abstract

Transient glutamate signaling often leads to long lasting and permanent alterations of a variety of cellular functions through particular membrane receptors in the brain. For elucidation of mechanisms underlying long-term consolidation of transient extracellular signals, we have examined expression and degradation of particular Fos family member proteins required for assembly to the nuclear transcription factor activator protein-1 in this study. Transcription factors could modulate the activity of RNA polymerase II responsible for the formation of mRNA from genomic DNA in the nucleus and therefore regulate de novo synthesis of particular target functional proteins. Mice were intraperitoneally injected with 100 mg/kg N-methyl-D-aspartic acid (NMDA) or 40 mg/kg kainic acid (KA), followed by homogenization of hippocampus in the presence of different protease and phosphatase inhibitors 2 h after administration, and subsequent preparation of nuclear and cytosolic fractions. The systemic administration of both NMDA and KA induced marked expression of particular Fos family members, including c-Fos and Fra-2 proteins, in hippocampal nuclear and cytosolic fractions. Incubation at 30 degrees C for 1 to 18 h led to differential degradation profiles of each Fos family member protein in nuclear fractions in a manner peculiar to the individual excitants. Degradation rate was also affected by dialysis and subsequent addition of inhibitors for phosphatases and proteases. These results suggest that in vivo NMDA and KA signals may additionally modulate the activity of heterologous machineries responsible for breakdown of each Fos family member in a unique manner in nuclear fractions, rather than cytosolic fractions, of murine hippocampus.