title: Real-time visualization of caspase-3 activation by fluorescence resonance energy transfer (FRET).
abstract: As apoptosis occurs via a complex signaling cascade that is tightly regulated at multiple cell points, different methods exist to evaluate the activity of the proteins involved in the intracellular apoptotic pathways and the phenotype of apoptotic neurons. Detention of the activity of the enzyme caspase-3, the key executioner caspase in programmed cell death, by laser scanning confocal fluorescence microscopy and the fluorescence resonance energy transfer technology is an alternative approach to classical standard techniques, such as Western blotting, activity assays, or histological techniques, and allows working with both fixed and living cells. This technique combined with the organotypic culture approach ex vivo represents a valid tool for the study of the mechanisms of neuronal survival /death and neuroprotection.
authors.affiliation: Department of Veterinary Sciences, University of Turin, Via Leonardo da Vinci 44, Turin, Grugliasco, 10095, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Alasia
authors.forename: Carolina
authors.initials: C
authors.lastname: Cocito
authors.forename: Adalberto
authors.initials: A
authors.lastname: Merighi
authors.forename: Laura
authors.initials: L
authors.lastname: Lossi
mesh_terms: D017209:Apoptosis / Q000235:genetics*; D053148:Caspase 3 / Q000235:genetics / Q000302:isolation & purification*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000235:genetics; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D018613:Microscopy, Confocal; D008967:Molecular Biology / Q000379:methods; D009474:Neurons / Q000378:metabolism / Q000648:ultrastructure*
chemical_list: D053148:Caspase 3