Multiparameter analysis of apoptosis in puromycin-treated Saccharomyces cerevisiae

Barbara Citterio, Maria Cristina Albertini, Lina Ghibelli, Elisabetta Falcieri, Michela Battistelli, Barbara Canonico, Marco B. L. Rocchi, Laura Teodori, Maurizio Ciani, Elena Piatti

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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.
Original languageEnglish
Pages (from-to)773 - 780
Number of pages8
JournalArchives of Microbiology
Volume197
Issue number6
DOIs
Publication statusPublished - 25 Aug 2015
Externally publishedYes

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All Science Journal Classification (ASJC) codes

  • Microbiology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Citterio, B., Albertini, M. C., Ghibelli, L., Falcieri, E., Battistelli, M., Canonico, B., ... Piatti, E. (2015). Multiparameter analysis of apoptosis in puromycin-treated Saccharomyces cerevisiae. Archives of Microbiology, 197(6), 773 - 780. https://doi.org/10.1007/s00203-015-1110-7