Minute kinetics of proapoptotic proteins

BAX and Smac/DIABLO in living tumor cells revealed by homeostatic confocal microscopy

Michal Marek Godlewski*, Magdalena Gorka, Monika Lamparska-Przybysz, Tomasz Motyl

*Corresponding author for this work

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Traditional methods of visualization and analysis based on fixed cell populations treated with the drug for a different time give the limited possibility of time-sequence analysis. In time-lapse microscopy where the whole cell is observed regardless to intracellular structure, precise localization of events and differentiation between colocalization and overlapping of the fluorescence is impossible. Furthermore prolonged experiments with living cells increased the influence of improper environmental conditions. Homeostatic confocal microscopy gives an exceptional insight into minute pattern of changes occurring in the same living cell maintained in stable conditions during whole experimental period. It is built on a confocal system equipped with the homeostatic chamber providing constant, monitored heating and moisturized, CO2-enriched atmosphere during long period observations. In the present study 2D/time and 4D homeostatic confocal microscopy were applied for analysis of minute pattern of changes occurring at the mitochondria. The release of Smac/DIABLO from mitochondria in tumor cells under the apoptogenic stimulus, consist of two phases: the first immediately after drug administration, and the major second one after 15 min. Furthermore the time-pattern of BAX translocation to the mitochondria and Smac/DIABLO release coincide, suggesting that the release of Smac/DIABLO is correlated with BAX translocation to the mitochondria.

Original languageEnglish
Pages (from-to)141-153
Number of pages13
JournalCytotechnology
Volume45
Issue number3
DOIs
Publication statusPublished - 2004

Keywords

  • BAX
  • Homeostatic confocal microscopy
  • Mitochondria
  • Smac/DIABLO
  • Time-pattern analysis

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