1. Changes in free intracellular calcium concentrations ([Ca2+]i) were measured in the nucleus and perinuclear regions of cultured rat hippocampal neurons using either fura-2 or fluo-3 calcium indicators. 2. Brief application of glutamate caused a transient rise of [Ca2+]i in all cell compartments, which recovered to pre-drug levels in all but the nuclear region. The new, higher level of nuclear calcium ([Ca2+]n) was sustained for as long as the cell was monitored. 3. The new level of [Ca2+]n was dependent on the magnitude of the calcium transient, and was higher in older cells in culture, but it did not affect responses to subsequent applications of glutamate. 4. The sustained elevation of [Ca2+]n was prevented by drugs which affect calcium stores (caffeine, ryanodine and Ruthenium Red), indicating that an extranuclear calcium store interacts with [Ca2+]n.