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Direct analysis of vescicular dynamics in neuronal and non-neuronal cells by a novel tool : dottorato di ricerca in fisiologia : tesi di dottorato

Ripamonti, Maddalena <1978- >

Tesi o dissertazioni - 2008

Maddalena Ripamonti Abstract: A fascinating aim is the possibility to visualize the dynamics of intracellular vesicular compartments as synaptic vesicles (SSVs). In the last years many tools have been developed to this use but, each one, has many limitations the most important of which is poor signal-to-noise ratio. We have developed a family of new tools highly specific for vesicular compartments in living cells. These tools are fusion constructs based on the scaffold of vesicular protein VAMP2. We inserted at the intralumenal end a peptide cassette containing an acidic sequence recognized with high affinity and specificity by a small (4,7 KDa) fluorescent basic peptide. When an exo-endocytotic cycle occurs, the intralumenal portion of these proteins is exposed to the extracellular environment and binds basic peptide molecules present in the extracellular medium. Expressing our probe in both neuronal and non-neuronal cells I could study the fate of vesicular compartments, or of individual synaptic boutons in living conditions, with a specificity and resolution never achieved before [...]
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