Repeated NMDA receptor activation induces distinct intracellular calcium changes in subpopulations of striatal...

The mechanisms underlying long-term calcium changes evoked by excitatory amino acids have not been previously examined in striatal neurons. Fura-2 fluorescence measurements were used to examine intracellular calcium concentration ([Ca2+]i) changes due to repeated N-methyl-D-aspartate (NMDA) receptor activation, in primary cultures of murine striatal neurons. Three applications of 200 microM NMDA (for 2 min, each application separated by 7 min), in 0 magnesium-containing artificial cerebral spinal fluid, elicited three distinct responses. In 50 +/- 8% of the NMDA-responsive neurons, no persistent increases in [Ca2+]i (final [Ca2+]i < or = 150% baseline) were observed, while in 33 +/- 7% and 17 +/- 3% of the cells, sustained (peak response > final [Ca2+]i > 150% baseline) and uncontrolled increases (final [Ca2+]i > or = peak response) were observed, respectively. NMDA-responsive neurons that were intensely immunoreactive for the calcium binding protein calbindin-D28k never exhibited uncontrolled increases in [Ca2+]i. Removal of extracellular Ca2+ significantly attenuated sustained, but not uncontrolled, increases in [Ca2+]i; sustained increases in some neurons were also attenuated by application of verapamil (100 microM) or MK-801 (1 microM). Pre-treatment of striatal neurons with the protein kinase C blocker sphingosine (20 microM), virtually eliminated the development of sustained or uncontrolled increases in [Ca2+]i. These findings suggest that specific intracellular mechanisms regulate the distinct [Ca2+]i responses of subpopulations of striatal neurons to repeated NMDA receptor activation.