Dynamic changes in the intracellular free Ca2+ concentration ([Ca2+]i) were studied in hippocampal cultured neurons using fluorescent Ca(2+)-indicator dye Indo-1 and somatic whole-cell recordings. During the tetanus stimulation Ca(2+)-transient increased their amplitude up to a steady-state level during repetitive stimulation. We identified two groups of neurons based on Ca-signal dynamics after the end of stimulation: the first group (n = 24) with the monoexponential decay of [Ca2+]i direct after the end of the tetanus; the second group (n = 32) with the monoexponential delayed [Ca2+]i decay after the end of the tetanus, the duration of delay varied from 1 to 27 s and depended on duration and frequency of stimulation. Peak amplitudes of Ca(2+)-transients were statistically different between the first (1820 +/- 195 nM, n = 24) and the second (2618 +/- 165 nM, n = 23) groups. A linear dependence between decay time constant and frequency of stimulation was found for the second group of neurons only. In all cases when the delayed decay was observed the decay time constant changed reliably after emergence of delayed decay; the average rise made up 41 +/- 8%. We suggest dynamic changes and essential rise in the intracellular free Ca2+ concentration arise from the presence of intracellular low-affinity buffer. This statement is to be further tested using pharmacological approach.