USE OF A MULTIWELL FLUORESCENCE SCANNER WITH PROPIDIUM IODIDE TO ASSESS NMDA MEDIATED EXCITOTOXICITY IN RAT CORTICAL NEURONAL CULTURES

Glutamate mediated excitotoxicity is a major area of experimentation d ue to the potential for prevention of morbidity and brain damage assoc iated with stroke and brain trauma. We have developed a simple rapid m ethod to study excitotoxicity in primary cortical neuronal cultures us ing propidium iodide (PI) fluorescence read by a multiwell fluorescenc e scanner. Transient (25 min) or continuous N-methyl-D-aspartate (NMDA ) treatment led to progressive neuronal death over 24 h that was block ed by 1 mu M MK-801, 10 mu M ifenprodil, and 200 mM ethanol. Results w ith PI fluorescence were identical to those found using the lactate de hydrogenase (LDH) release and trypan blue staining assays of excitotox icity. This method provides a simple rapid means to test the effects o f drugs during glutamate excitotoxicity and to do accurate time course experiments of delayed neuronal death. (C) 1997 Elsevier Science Irel and Ltd.