Apoptosis

, Volume 10, Issue 6, pp 1317–1331 | Cite as

Biphasic behavior of changes in elemental composition during staurosporine-induced apoptosis

  • F. Arrebola
  • J. Cañizares
  • M. A. Cubero
  • P. V. Crespo
  • A. Warley
  • E. Fernández-Segura
Article

Abstract

Although the identification of events that occur during apoptosis is a fundamental goal of apoptotic cell death research, little is know about the precise sequence of changes in total elemental composition during apoptosis. We evaluated total elemental composition (Na, Mg, P, Cl, S, and K) in relation to molecular and morphological features in human U937 cells induced to undergo apoptosis with staurosporine, an intrinsic pathway activator. To evaluate total elemental content we used electron probe X-ray microanalysis to measure simultaneously all elements from single, individual cells. We observed two phases in the changes in elemental composition (mainly Na, Cl and K). The early phase was characterized by a decrease in intracellular K (P < 0.001) and Cl (P < 0.001) content concomitant with cell shrinkage, and preceded the increase in proteolytic activity associated with the activation of caspase-3. The later phase started with caspase-3 activation, and was characterized by a decrease in the K/Na ratio (P < 0.001) as a consequence of a significant decrease in K and increase in Na content. The inversion of intracellular K and Na content was related with the inhibition of Na+/K+ ATPase. This later phase was also characterized by a significant increase (P < 0.001) in intracellular Cl with respect to the early phase. In addition, we found a decrease in S content and an increase in the P/S ratio. These distinctive changes coincided with chromatin condensation and DNA fragmentation. Together, these findings support the concept that changes in total elemental composition take place in two phases related with molecular and morphological features during staurosporine-induced apoptosis.

Keywords

apoptosis apoptotic volume decrease caspases chlorine electron probe X-ray microanalysis Na+/K+ ATPase PARP potassium sodium 

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Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  • F. Arrebola
    • 1
  • J. Cañizares
    • 1
    • 2
  • M. A. Cubero
    • 1
  • P. V. Crespo
    • 1
    • 2
  • A. Warley
    • 3
  • E. Fernández-Segura
    • 1
    • 2
    • 4
  1. 1.Department of Histology, Faculty of MedicineUniversity of GranadaGranadaSpain
  2. 2.Institute of NeurosciencesUniversity of GranadaGranadaSpain
  3. 3.CUI King's College LondonLondonUK
  4. 4.Department of Histology, Faculty of MedicineUniversity of GranadaGranadaSpain

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