[Measurement of free intracellular Ca2+ concentration in cultivated hippocampal neurons]

The cascade of pathobiochemical reactions caused by cerebral ischemia is complex and not completely understood. One important factor seems to be the disturbed calcium homeostasis. Measurement of cytosolic-free Ca2+ concentration [Ca2+]i is possible with the Ca(2+)-sensitive dye fura-2. Using this technique it can be investigated if, and how, neuroprotective drugs influence the cellular calcium homeostasis. In hippocampal neurons K+ or glutamate, added to the buffer solution, evoked elevations of [Ca2+]i in a concentration-dependent manner. We demonstrated that the calcium antagonist nimodipine or the non-competitive NMDA receptor antagonist dizocilpine added to the buffer solution could inhibit concentration-dependently the elevation of [Ca2+]i. The measurement of [Ca2+]i can be used for studies on the mechanism of action as well as for screening purposes of neuroprotective drugs.