title: Multiparameter analysis of apoptosis in puromycin-treated Saccharomyces cerevisiae.
abstract: In Saccharomyces cerevisiae, a typical apoptotic phenotype is induced by some stress factors such as sugars, acetic acid, hydrogen peroxide, aspirin and age. Nevertheless, no data have been reported for apoptosis induced by puromycin, a damaging agent known to induce apoptosis in mammalian cells. We treated S. cerevisiae with puromycin to induce apoptosis and evaluated the percentage of dead cells by using Hoechst 33342 staining, transmission electron microscopy (TEM) and Annexin V flow cytometry (FC) analysis. Hoechst 33342 fluorescence images were processed to acquire parameters to use for multiparameter analysis [and perform a principal component analysis, (PCA)]. Cell viability was evaluated by Rhodamine 123 (Rh 123) and Acridine Orange microscope fluorescence staining. The results show puromycin-induced apoptosis in S. cerevisiae, and the PCA analysis indicated that the increasing percentage of apoptotic cells delineated a well-defined graph profile. The results were supported by TEM and FC. This study gives new insights into yeast apoptosis using puromycin as inducer agent, and PCA analysis may complement molecular analysis facilitating further studies to its detection.
authors: Department of Biomolecular Sciences, University of Urbino "Carlo Bo", Urbino, Italy.
authors: Barbara
authors: B
authors: Citterio
authors: Maria Cristina
authors: MC
authors: Albertini
authors: Lina
authors: L
authors: Ghibelli
authors: Elisabetta
authors: E
authors: Falcieri
authors: Michela
authors: M
authors: Battistelli
authors: Barbara
authors: B
authors: Canonico
authors: Marco B L
authors: MB
authors: Rocchi
authors: Laura
authors: L
authors: Teodori
authors: Maurizio
authors: M
authors: Ciani
authors: Elena
authors: E
authors: Piatti
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D005434:Flow Cytometry; D046529:Microscopy, Electron, Transmission; D025341:Principal Component Analysis; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D011691:Puromycin / Q000494:pharmacology*; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000502:physiology
chemical_list: D011500:Protein Synthesis Inhibitors; D011691:Puromycin