Calbindin-28K (CaBP) is a calcium-binding protein widely distributed in the brain. This protein appears to be involved in the sequestration and the translocation of intracellular free calcium. In this study, we have examined the distribution pattern of the structures immunoreactive for CaBP in the hippocampal formation from slices of young (4 months) and aged (24–27 months) rats previously submitted to electrophysiological measurements. We demonstrated a marked loss in the number of pyramidal cells immunoreactive for CaBP in aged rats as compared to young rats. A consistent decrease in the staining intensity was also revealed by optical density measurements. Some experiments have suggested that calcium homeostasis is modified in hippocampal neurons of aged rats. The loss of CaBP-like immunoreactivity (CaBP-LI) labelling could result from an increase in intracellular calcium concentrations. To support this hypothesis, we showed that in young rats (i) the CaBP-LI was enhanced in pyramidal neurons when the slice was preincubated in a calcium-free medium, and (ii) CaBP-LI was strongly decreased when the slice was preincubated in a high-calcium medium (5 mM) and when the entry of calcium into the cell was increased by a short application of an excitatory amino acid in the medium. Our results suggest that the loss of CaBP-LI in the hippocampus of aged rats could be due to an increase in intracellular calcium concentration. Preliminary observations of hippocampal slices at different times after induction of long-term potentiation (LTP) failed to show significant changes in CaBP immunoreactivity, suggesting that this calcium-binding protein is not directly involved in LTP processes.