Protoplasma

, Volume 198, Issue 3–4, pp 163–169 | Cite as

A structural analysis of cytoskeleton components during the execution phase of apoptosis

  • R. Atencia
  • M. García-Sanz
  • G. Pérez-Yarza
  • A. Asumendi
  • E. Hilario
  • J. Aréchaga
Article
Received:
Accepted:

Summary

During the execution phase of apoptosis, the cell undergoes a set of morphological changes which reveal the activation of a complex machinery leading the cell to its disruption into small, spherical, membrane-bounded fragments called apoptotic bodies. In the present study, we have focused on the implications of the micro-filament network in the early stages of the active phase of apoptosis. By using confocal microscopy, we have analysed the location of the actin microfilaments and two actin-binding proteins, α-actinin and myosin, in F9 embryonal carcinoma cells undergoing apoptosis during the stages previous to their fragmentation. Our results show that these proteins locate in the centre of the disrupting cell and form a three-dimensional structure which suggests the existence of a fully functional contractile system involved in the fragmentation of the cell and the formation of apoptotic bodies.

Keywords

Actin Actin-binding proteins Apoptosis F9 embryonal carcinoma cells 

Abbreviations

CI-II

calpain inhibitor II

CD

cytochalasin D

CSLM

confocal scanning laser microscopy

EC

embryonal carcinoma

FALS

forward angle side scattered

FCS

fetal calf serum

FITC

fluorescein isothyocyanate

ISS

integrated side scattered

PBS

phosphate buffered saline: PI propidium iodide

RA

retinoic acid

TEM

transmission electron microscopy

This is a preview of subscription content, log in to check access.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Arends MJ, Wyllie AH (1991) Apoptosis: mechanisms and roles in pathology. Int Rev Exp Pathol 52: 223–254Google Scholar
  2. Atencia R, García-Sanz M, Unda F, Aréchaga J (1994) Apoptosis during retinoic acid-induced differentiation of F9 embryonal carcinoma cells. Exp Cell Res 214: 663–667Google Scholar
  3. Brancolini C, Benedetti M, Schneider C (1995) Microfilament reorganization during apoptosis: the role of Gas2, a possible substrate for ICE-like proteases. EMBO J 21: 5179–5190Google Scholar
  4. Cooper JA (1987) Effects of cytochalasin and phalloidin on actin. J Cell Biol 105: 1473–1478Google Scholar
  5. Cotter JA, Lennon SV, Glynn JM, Green DR (1992) Microfilament disrupting agents prevent the formation of apoptotic bodies in tumour cells. Cancer Res 52: 997–1005Google Scholar
  6. Earnshaw WC (1995) Apoptosis: lessons from in vitro systems. Trends Cell Biol 5: 217–220Google Scholar
  7. Fesus L, Davies PJA, Piacentini M (1991) Apoptosis: molecular mechanisms in programmed cell death. Eur J Cell Biol 56: 170–177Google Scholar
  8. Fox JEB, Boyles JK (1988) The membrane skeleton: a distinct structure that regulates the function of cells. Bioessays 8: 14–18Google Scholar
  9. —, Reynolds CC, Morrow JS, Phillips DR (1987) Spectrin is associated with membrane-bound actin filaments in platelets and is hydrolyzed by endogenous Ca2+-dependent protease during platelet activation. Blood 69: 537–545Google Scholar
  10. García-Sanz M, Atencia R, Pérez-Yarza G, Asumendi A, Hilario E, Aréchaga J (1996) Cytoarchitectural changes during retinoic acid-induced apoptosis in F9 embryonal carcinoma cells. Int J Dev Biol 40 Suppl 1: 195–196Google Scholar
  11. Kerr JFR, Wyllie AH, Currie AR (1972) Apoptosis: a basic biological phenomenon with wide range implications in tissue kinetics. Br J Cancer 26: 239–257Google Scholar
  12. Kumar S (1995) ICE-like proteases in apoptosis. Trends Biochem Sci 20: 198–202Google Scholar
  13. Lazebnik YA, Takahasi A, Poirier GG, Kaufmann SH, Earnshaw (1995) Characterization of the execution phase of apoptosis in vitro using extracts from condemned-phase cells. J Cell Sci Suppl 19: 41–49Google Scholar
  14. Martin SJ, Green DR (1995) Protease activation during apoptosis: death by a thousand cuts? Cell 82: 349–352Google Scholar
  15. —, O'Brien GA, Nishioka WK, McGahon AJ, Mahboubi A, Saido TC, Green DR (1995) Proteolysis of fodrin (non-erythroid spectrin) during apoptosis. J Biol Chem 270: 6425–6428Google Scholar
  16. Miura M, Zhu H, Rotello R, Hartwieg EA, Yuan J (1993) Induction of apoptosis in fibroblasts by IL-1β-converting enzyme, a mammalian homolog of theC. elegans cell death geneced-3. Cell 82: 349–352Google Scholar
  17. Naora H, Naora H (1995) Differential expression patterns of β-actin mRNA in cells undergoing apoptosis. Biochem Biophys Res Commun 211: 491–496Google Scholar
  18. Onji T, Takagi M, Shibata N (1987) Calpain abolishes the effect of filamin on the actomyosin system in platelets. Biochim Biophys Acta 912: 283–286Google Scholar
  19. Schollmeyer JE (1988) Calpain II involvement in mitosis. Science 240: 911–913Google Scholar
  20. Squier MKT, Miller ACK, Malkinson AM, Cohen JJ (1994) Calpain activation in apoptosis. J Cell Physiol 159: 229–237Google Scholar
  21. Unda F, García-Sanz M, Atencia R, Hilario E, Aréchaga J (1994) Co-expression of laminin and a 67 kDa laminin-binding protein in teratocarcinoma embryoid bodies. Int J Dev Biol 38: 121–126Google Scholar
  22. Wyllie AH (1981) Cell death: a new classification separating apoptosis from necrosis. In: Bowen I, Lochshin RA (eds) Cell death in biology and pathology. Chapman and Hall, London, pp 9–23Google Scholar
  23. Yuan J (1995) Molecular control of life and death. Curr Opin Cell Biol 7: 211–214Google Scholar
  24. Zhivotovsky B, Gham A, Ankarcrona M, Nicotera PL, Orrenius S (1995) Multiple proteases are involved in thymocyte apoptosis. Exp Cell Res 221: 404–412Google Scholar

Copyright information

© Springer-Verlag 1997

Authors and Affiliations

  • R. Atencia
    • 1
  • M. García-Sanz
    • 1
  • G. Pérez-Yarza
    • 1
  • A. Asumendi
    • 1
  • E. Hilario
    • 1
  • J. Aréchaga
    • 1
  1. 1.Departamento de Biología Celular, Facultad de MedicinaUniversidad del País VascoVizcayaSpain

Personalised recommendations