Within neurites of nearly all cultured hippocampal neurons, transient ATP depletion rapidly inducesrod- shaped structures composed primarily of actin and ADF/cofilin (AC). Rod formation, which sequesters a portion of the actin but virtually all of the AC, is transiently beneficial to the stressed neuron because it spares ATP associated with actin turnover. However, rods can completely occlude the neurite, blocking transport and causing distal neurite withering. Rods are prominent features of Alzeimer's disease (AD) brain but not of control human brain lacking amyloid plaques. Similar structures are found in brains of animals with Niemann-Pick disease typed (NPC1) and of transgenic mice (Tg2576) expressing mutant human amyloid precursor protein (APP). In cultured neurons and mouse brain slices, rods are induced by ischemia, peroxide, NO, and excitotoxic glutamate. In up to 20% of hippocampal neurons, whether from region CA1 or CAS, the AD amyloid beta peptide (Ab) also induces rods: induction is dose-dependent, occuring within 6 h after treatment and plateauing 12-24 h later. As little as 10 nM of Ab oligomer has a significant effect compared to the scrambled peptide control. The nature of the sensitivity of only a subset of neurons to Ab will be explored. Rods block vesicular transport of APP. APP-containing vesicles accumulate at the ends and sides of rods. Within these stalled vesicles is beta-secretase cleaved APP, suggesting that these may be sites of Ab production and/or conversion into more damaging conformers. Taken together, these results suggest a model for AD in which neuronal stress, including Ab formed in familial AD, induces rods that stall vesicle transport and increase toxic Ab, thus inducing rods in neighboring cells. Such a model could explain the formation of amyloid plaques, which would enlarge around the initial site of injury. Using cell culture and organotypic brain slices, we will determine: 1) what activities of cofilin are required for rod formation; 2) if mutations in AC can be identified that prevent rod formation; 3) if rods promote the production or oligomerization of Ab; 4) what makes a subset of neurons sensitive to Ab: and 5) how organotypic brain slices can be used as a model to study where rods form and how they disrupt synapses. Relevance to public health: AD dramatically impacts life quality of senior Americans, affecting 25% of those > 85. This proposal tests a new hypothesis for AD progression and identifies possible sites for targeted intervention.
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