Converging evidence suggests that the accumulation of cerebral amyloid beta-protein (Abeta) in Alzheimer's disease (AD) reflects an imbalance between the production and degradation of this self-aggregating peptide. Upregulation of proteases that degrade Abeta thus represents a novel therapeutic approach to lowering steady-state Abeta levels, but the consequences of sustained upregulation in vivo have not been studied. Here we show that transgenic overexpression of insulin-degrading enzyme (IDE) or neprilysin (NEP) in neurons significantly reduces brain Abeta levels, retards or completely prevents amyloid plaque formation and its associated cytopathology, and rescues the premature lethality present in amyloid precursor protein (APP) transgenic mice. Our findings demonstrate that chronic upregulation of Abeta-degrading proteases represents an efficacious therapeutic approach to combating Alzheimer-type pathology in vivo.

Amyloid-␤ protein (A␤) aggregates in the brain to form senile plaques. By using thioflavin T, a dye that specifically binds to fibrillar structures, we found that metals such as Zn(II) and Cu(II) normally inhibit amyloid ␤-aggregation. Another method for detecting A␤, which does not distinguish the types of aggregates, showed that these metals induce a non-␤-sheeted aggregation, as reported previously. Secondary structural analysis and microscopic studies revealed that metals induced A␤ to make non-fibrillar aggregates by disrupting ␤-sheet formation. These non-fibrillar A␤ aggregates displayed much weaker Congo Red birefringence, and in separate cell culture experiments, were less toxic than self ␤-aggregates, as demonstrated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. The toxicity of soluble A␤ was enhanced in the presence of Cu(II), which suggests the previously hypothesized role of A␤ in generating oxidative stress. Finally, under an acidic condition, similar to that in the inflammation associated with senile plaques, ␤-aggregation was robustly facilitated at one specific concentration of Zn(II) in the presence of heparin. However, because a higher concentration of Zn(II) virtually abolished this abnormal phenomenon, and at normal pH any concentrations strongly inhibit ␤-aggregation and its associated cytotoxicity, including its anti-oxidative nature we suggest that Zn(II) has an overall protective effect against ␤-amyloid toxicity. Amyloid-␤ protein (A␤)1 is one of the main components of senile plaques, a pathological hallmark of Alzheimer's disease (AD) (1, 2). Although A␤ is undisputedly associated with the pathology of AD, it is still an open question as to what specific aspects of A␤ and its processing are the important variables in the pathophysiology of the disease. For example, fibrillar A␤, but not non-amyloidogenic, amorphous aggregates of A␤, was reported to cause neuronal cell death in primary rat hippocampal cultures (3), and soluble monomeric species of A␤ are relatively nontoxic as compared with fibrillar A␤ (4). Thus, these in vitro studies suggest that the degree of ␤-aggregation is particularly important for neurotoxicity to occur (5-8). However, many controversial results from in vivo studies have been reported concerning the pathological role of plaque formation in AD. Irizarry et al. (9) reported that transgenic (TG) mice expressing human A␤ failed to exhibit neuronal loss despite depositing substantial amounts of A␤. On the other hand, TG mice that express Swedish mutant amyloid precursor protein (APP) formed plaques that were detected by both an anti-APP antibody and a ␤-sheet specific dye (10). Moreover, these APP TG mice also displayed memory deficits. Taken together, these results indicate that, although the plaque assembly process may require further investigation, amyloid ␤-aggregation certainly is an essential event in the pathogenesis of AD.Based on these lines of evidence, the search for a compound that interrupts ␤-aggregation and thus protects agai...

Patients with AD had less density of retinal microvascular networks than controls. Our findings suggest the presence of retinal microvascular dysfunction in AD.

Aberrant processing of the amyloid precursor protein (APP) and the subsequent accumulation of amyloid ␤ (A␤) peptide has been widely established as a central event in Alzheimer's disease (AD) pathogenesis. The sequential cleavage steps required for the generation of A␤ are well outlined; however, there is a relative dearth of knowledge pertaining to signaling pathways and molecular mechanisms that can modulate this process. Here, we demonstrate a novel role for p25/cyclin-dependent kinase 5 (Cdk5) in regulating APP processing, A␤ peptide generation, and intraneuronal A␤ accumulation in inducible p25 transgenic and compound PD-APP transgenic mouse models that demonstrate deregulated Cdk5 activity and a neurodegenerative phenotype. Induction of p25 resulted in enhanced forebrain A␤ levels before any evidence of neuropathology in these mice. Intracellular A␤ accumulated in perinuclear regions and distended axons within the forebrains of these mice. Evidence for modulations in axonal transport or ␤-site APP cleaving enzyme 1 protein levels and activity are presented as mechanisms that may account for the A␤ accumulation caused by p25/Cdk5 deregulation. Collectively, these findings delineate a novel pathological mechanism involving aberrant APP processing by p25/Cdk5 and have important implications in AD pathogenesis.