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 due to the potential for prevention of morbidity and brain damage associated with stroke and brain trauma. We have developed a simple rapid method to study excitotoxicity in primary cortical neuronal cultures using propidium iodide (PI) fluorescence read by a multiwell fluorescence scanner. Transient (25 min) or continuous N-methyl-D-aspartate (NMDA) treatment led to progressive neuronal death over 24 h that was blocked by 1 microM MK-801, 10 microM ifenprodil, and 200 mM ethanol. Results with PI fluorescence were identical to those found using the lactate dehydrogenase (LDH) release and trypan blue staining assays of excitotoxicity. This method provides a simple rapid means to test the effects of drugs during glutamate excitotoxicity and to do accurate time course experiments of delayed neuronal death.
Rudolph
J G
JG
Department of Pharmacology and Therapeutics, University of Florida, Gainesville 32610-0267, USA.
Lemasters
J J
JJ
Crews
F T
FT
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D018690:Excitatory Amino Acid Agonists / Q000633:toxicity*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D016202:N-Methylaspartate / Q000633:toxicity*; D009474:Neurons / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D011419:Propidium; D051381:Rats
D018690:Excitatory Amino Acid Agonists; D011419:Propidium; D016202:N-Methylaspartate; D007770:L-Lactate Dehydrogenase