The effect of 'simulated ischemia', i.e., combined anoxia and substrate deprivation, was studied in 1- and 3-week-old (i.e., immature and mature) primary cultures of mouse astrocytes. Cell survival, as indicated by retention of the high-molecular cytosolic protein lactate dehydrogenase was compared with retained high-energy phosphate compounds (ATP and phosphocreatine). A previously established longer survival of the immature cells during the metabolic insult was confirmed and found to correlate with a more complete maintenance of high-energy phosphates. However, in both the mature and immature cells, no death occurred as long as the ATP content remained at or above 25% of its control value. ATP concentrations below 10% of control were accompanied by almost complete cell death in both age groups. Thus, the better survival of immature astrocytes during simulated ischemia is correlated with better maintenance of the levels of high-energy phosphates and, regardless of age, cell death occurs only once a critically 'low' threshold of ATP has been reached.