Granule cells in a dissociated neuro-glial cell culture of cerebellum when exposed to ouabain (10 33 M) for 25 min apparently swell, increase their [Ca 2+ ] i with obvious depolarization of the mitochondrial membrane. In 3 h after ouabain was omitted from the solution, 62 þ 3% of granule cells had pycnotic nuclei. The supplement of a solution with competitive specific antagonist of NMDA receptors, L-2-amino-7-phosphonoheptanoate (10 34 M, APH) together with ouabain prevented cells from swelling, mitochondrial deenergization, neuronal death and increase of [Ca 2+ ] i . These data suggest that cellular Na + /K + -ATPase inactivation in neuro-glial cell cultures of cerebellum leads to glutamate (Glu) accumulation, hyperstimulation of glutamate receptors, higher Ca 2+ and Na + influxes into the cells through the channels activated by Glu. This process leads to cell swelling, mitochondrial deenergization and death of granule cells. Possibly, the decrease of Na + /K + -ATPase activity in brain cells can lead to the onset of at least some chronic neurological disorders.z 1999 Federation of European Biochemical Societies.