The neurotoxicity of beta-amyloid protein (AbetaP) is implicated in the etiology of Alzheimer's disease. We previously have demonstrated that AbetaP forms Ca(2+)-permeable pores on neuronal membranes, causes a marked increase in intracellular calcium level, and leads to neuronal death. Here, we investigated in detail the features of AbetaP-induced changes in intracellular Ca(2+) level in primary cultured rat hippocampal neurons using a multisite Ca(2+)-imaging system with fura-2 as a fluorescent probe. Only a small fraction of short-term cultured hippocampal neurons (ca 1 week in vitro) exhibited changes in intracellular Ca(2+) level after AbetaP exposure. However, AbetaP caused an acute increase in intracellular Ca(2+) level in long-term cultured neurons (ca 1 month in vitro). The responses to AbetaP were highly heterogeneous, and immunohistochemical analysis using an antibody to AbetaP revealed that AbetaP is deposited on some but not all neurons. Considering that the disruption of Ca(2+) homeostasis is the primary event in AbetaP neurotoxicity, substances that protect neurons from an AbetaP-induced intracellular Ca(2+) level increase may be candidates as therapeutic drugs for Alzheimer's disease. In line with the search for such protective substances, we found that the preadministration of neurosteroids including dehydroepiandrosterone, dehydroepiandrosterone sulfate, and pregnenolone significantly inhibits the increase in intracellular calcium level induced by AbetaP. Our results suggest the possible significance of neurosteroids, whose levels are reduced in the elderly, in preventing AbetaP neurotoxicity.
Keywords: aging; calcium homeostasis; channel; neurotoxicity; pore.