Regulation of N-methyl-D-aspartate receptor-mediated calcium transport and norepinephrine release in rat hippocampus synaptosomes by polyamines

Abstract

The role of polyamines (PA) synthesis in NMDA receptor-mediated 45Ca2+ fluxes and norepinephrine release was studied in rat hippocampal synaptosomes. NMDA (50 microM) caused a sharp (> 2-fold) transient increase in PA synthesis regulating enzyme, ornithine decarboxylase (ODC) activity with concomitant elevation in PA levels in the order putrescine > spermidine > spermine. ODC inhibitor, alpha-difluoromethylornithine (DFMO), and NMDA antagonist, 2-amino-5-phosphonovaleric acid (D-AP5), both blocked increases in ODC activity and PA levels. Activation of NMDA receptors induced a sharp (3 to 4-fold) and quick (15 seconds) increase in 45Ca2+ uptake by synaptosomes within 15 seconds of exposure at 37 degrees C. The efflux of 45Ca2+ and 3H-norepinephrine (NE) release at 22 degrees C from pre-loaded synaptosomes was also significantly (2 to 4-fold) enhanced by NMDA within 15 seconds. These NMDA receptor-mediated effects on calcium fluxes and NE release were blocked by NMDA receptor-antagonists (DAP-5 and MK-801) and PA synthesis inhibitor, DFMO and the DFMO inhibition nullified by exogenous putrescine. These observations establish that ODC/PA cascade play an important role in transduction of excitatory amino acid mediated signals at NMDA receptors.