Innocent_2010_Neuropharmacol_59_343

Key neuropathological hallmarks of Alzheimer's disease include the accumulation of amyloid-beta (Abeta), disruption of Ca(2+) homeostasis and neurodegeneration. However, the physical nature of the toxic Abeta species is controversial. Here, we examined the effect of aging on acute and chronic actions of Abeta peptides: changes in intracellular Ca(2+) and toxic responses, respectively. Acute application of Abeta(1-42) to PC12 cells potentiated KCl-evoked increases in Ca(2+), while chronic application decreased mitochondrial function with concomitant perturbation of membrane integrity and activation of apoptosis in PC12 cells, and reduced neurite length and synaptogenesis in rat cortical neurons. Both the acute and chronic effects of Abeta(1-42) were prevented by the anti-oligomerisation peptide D-KLVFFA, implicating oligomeric structures. The generation of a range of oligomeric species by aging Abeta(1-42) at 37 degrees C for different times was supported by thioflavin T fluorescence and atomic force microscopy. Abeta(1-42) aged for 24 h maximally potentiated KCl-evoked increases in Ca(2+), and this correlated with oligomers composed of 3-6 monomers, as judged by size exclusion filtration. Aging for 72 or 96 h, which generated fibrillar structures, was less efficacious. The Abeta(25-35) fragment that lacks the self-recognition element targeted by D-KLVFFA failed to potentiate KCl-evoked increases in Ca(2+). However, Abeta(25-35) was more efficacious than Abeta(1-42) at decreasing cellular functions when applied chronically. The acute and chronic effects of Abeta(1-42) also showed differential sensitivity to blockade of voltage operated Ca(2+) channels. These results suggest that the acute effects of Abeta(1-42) on Ca(2+) signals do not underpin the toxic responses measured, although both acute and chronic effects are promoted by small oligomeric species.