EXCITOTOXIC DEGENERATION IS INITIATED AT NONRANDOM SITES IN CULTURED RAT CEREBELLAR NEURONS

Prolonged stimulation of cultured cerebellar neurons by kainic acid (K A) leads to death of neurons first evident from the swelling of soma a nd neurites. Stimulation is accompanied by increases in [Ca2+](l) and [Na+](i) as monitored using digital imaging microfluorimetry. ''Blebs' ' tended to form on neurites with the highest increases in [Ca2+](i). Points of Ca2+ entry into neurites via glutamate-receptor-gated channe ls predicted where approximately 80% of blebs would form tens of minut es later. These sites were close to neurite intersections where there was a high likelihood of synaptic contacts and were enriched in mitoch ondria as revealed by rhodamine 123 staining. Ca2+, but not Na+ entry, produced a loss of mitochondrial potential. Prolonged KA, but not 50K , applications could fully dissipate the neuronal Na+ gradient. Recove ry of resting [Na+](i) was delayed by Ca2+ loading. We propose that bl ebs form at certain synaptic regions due to localized ionic fluxes and local Ca2+ overloading. Increased [Ca2+](l) may hamper restoration of normal [Na+](i) permitting local osmotic swelling as well as activati on of Ca2+-dependent enzymes and other processes. Na+ may slow, block, or reverse Na/Ca exchange and enhance swelling. These conditions coul d not be reproduced by global changes in ion concentrations produced b y Ca2+ or Na+ ionophores. The earliest stages of excitotoxicity thus a ppear to be manifestations of localized disruptions of ionic homeostas is mediated by Ca2+ overload and Na+ influx.