Fe³⁺ Facilitates Endocytic Internalization of Extracellular Aβ_1-42 and Enhances Aβ_1-42-Induced Caspase-3/Caspase-4...

Amyloid β (Aβ) peptide is a critical causative factor in Alzheimer's disease (AD) and of a variety of fragmented Aβ peptides Aβ_1-42 thought to exhibit the most neurotoxic effect. The present study investigated the effects of Fe³⁺ on Aβ_1-42 internalization and Aβ_1-42-induced caspase activation and neurotoxicity using mouse hippocampal slices and cultured PC-12 cells. Extracellularly applied Aβ_1-42 increased the cell-associated Aβ_1-42 levels in a concentration-dependent manner, and the effect was enhanced by adding Fe³⁺. Fe³⁺-induced enhancement of the cell-associated Aβ_1-42 levels was significantly inhibited by the endocytosis inhibitors dynasore and methyl-β-cyclodextrin. Aβ_1-42 reduced PC-12 cell viability in a concentration-dependent manner, and further reduction of the cell viability was obtained with Fe³⁺. Aβ_1-42-induced reduction of cell viability was not affected by A187, an antagonist of amylin-3 receptor. Aβ_1-42 activated caspase-3, caspase-4, and caspase-8 to a variety of degrees and Fe³⁺ further enhanced Aβ_1-42-induced activation of caspase-3 and caspase-4. Taken together, these results indicate that Fe³⁺ accelerates endocytic internalization of extracellular Aβ_1-42, enhances Aβ_1-42-induced caspase-3/caspase-4 activation, and promotes Aβ_1-42-induced neuronal cell death, regardless of amylin receptor.