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Alzheimer's disease (AD) is a neurodegenerative disorder characterized by progressive and irreversible loss of memory and other cognitive functions. Substantial evidence based on genetic, neuropathological and biochemical data has established the central role of beta-amyloid protein (betaAP) in this pathology. Although the precise etiology of AD is not well understood yet, strong evidence for some of the molecular events that lead to progressive brain dysfunction and neurodegeneration in AD has been afforded by identification of biochemical pathways implicated in the generation of betaAP, development of transgenic models exhibiting progressive disease pathology and by data on the effects of betaAP at the neuronal network level. However, the mechanisms by which betaAP causes cognitive decline have not been determined, nor is it clear if the degree of dementia correlates in time with the degree of neuronal loss. Hence, it is of interest to understand the biochemical processes involved in the mechanisms of betaAP-induced neurotoxicity and the mechanisms involved in electrophysiological effects of this protein on different parameters of synaptic transmission and on neuronal firing properties. In this review we analyze recent evidence suggesting a complex role of betaAP in the molecular events that lead to progressive loss of function and eventually to neurodegeneration in AD as well as the therapeutic implications based on betaAP metabolism inhibition.

The molecular characterization of extracellular vesicles (EVs) has revealed a great heterogeneity in their composition at a cellular and tissue level. Current isolation methods fail to efficiently separate EV subtypes for proteomic and functional analysis. The aim of this study was to develop a reproducible and scalable isolation workflow to increase the yield and purity of EV preparations. Through a combination of polymer‐based precipitation and size exclusion chromatography (Pre‐SEC), we analyzed two subsets of EVs based on their CD9, CD63 and CD81 content and elution time. EVs were characterized using transmission electron microscopy, nanoparticle tracking analysis, and Western blot assays. To evaluate differences in protein composition between the early‐ and late‐eluting EV fractions, we performed a quantitative proteomic analysis of MDA‐MB‐468‐derived EVs. We identified 286 exclusive proteins in early‐eluting fractions and 148 proteins with a differential concentration between early‐ and late‐eluting fractions. A density gradient analysis further revealed EV heterogeneity within each analyzed subgroup. Through a systems biology approach, we found significant interactions among proteins contained in the EVs which suggest the existence of functional clusters related to specific biological processes. The workflow presented here allows the study of EV subtypes within a single cell type and contributes to standardizing the EV isolation for functional studies.

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