Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Normal Hemopoiesis

Caspase activation is involved in early megakaryocyte differentiation but not in platelet production from megakaryocytes

Abstract

To elucidate whether caspase activation is involved in megakaryopoiesis, we characterized megakaryocytes (MKs) in vav-bcl-2 transgenic (Tg) mice, in which Bcl-2 is overexpressed in hematopoietic cells. To exclude the effect of splenomegaly in Tg mice on megakaryopoiesis, splenectomy was performed. After splenectomy, basal platelet counts in peripheral blood were not significantly different between Tg and wild-type (WT) mice. However, when experimental thrombocytopenia was induced by injecting 5-fluorouracil into splenectomized mice, overshoot of platelet counts during the recovery phase was hardly observed in Tg mice. Analyses of MK ploidy during the recovery phase showed that MKs less than 16 N ploidy were significantly decreased in Tg mice, suggesting that MK supply from progenitors is impaired. Supporting this, differentiation of CD34−/c-kit+/Sca-1+/Lineage− stem cells into MKs was significantly hampered in Tg mice, whereas megakaryocyte-erythroid progenitors (MEPs) normally differentiated into MKs. It suggests that differentiation into MKs is impaired in Tg mice before the stage of MEP. Furthermore, MK colony formation in WT cells was dose-dependently inhibited in the presence of a caspase inhibitor. Contrary, Bcl-2-overexpressing MKs showed normal ability for in vitro platelet production. We thus believe that caspase activation is involved in the differentiation of progenitors into megakaryocytic lineage but not in platelet production.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  1. Shivdasani RA . Molecular and transcriptional regulation of megakaryocyte differentiation. Stem Cells 2001; 19: 397–407.

    Article  CAS  Google Scholar 

  2. Pang L, Weiss MJ, Poncz M . Megakaryocyte biology and related disorders. J Clin Invest 2005; 115: 3332–3338.

    Article  CAS  Google Scholar 

  3. Italiano Jr JE, Lecine P, Shivdasani RA, Hartwig JH . Blood platelets are assembled principally at the ends of proplatelet processes produced by differentiated megakaryocytes. J Cell Biol 1999; 147: 1299–1312.

    Article  CAS  Google Scholar 

  4. Junt T, Schulze H, Chen Z, Massberg S, Goerge T, Krueger A et al. Dynamic visualization of thrombopoiesis within bone marrow. Science 2007; 317: 1767–1770.

    Article  CAS  Google Scholar 

  5. Italiano JE, Hartwig JH . Megakaryocyte development and platelet formation. In: Michelson AD (ed). Platelets 2nd edn. Academic Press: Burlington, MA, 2007, pp 23–44.

    Chapter  Google Scholar 

  6. Zauli G, Vitale M, Falcieri E, Gibellini D, Bassini A, Celeghini C et al. In vitro senescence and apoptotic cell death of human megakaryocytes. Blood 1997; 90: 2234–2243.

    CAS  PubMed  Google Scholar 

  7. Sanz C, Benet I, Richard C . Antiapoptotic protein Bcl-xL is up-regulated during megakaryocytic differentiation of CD34+ progenitors but is absent from senescent megakaryocytes. Exp Hematol 2001; 29: 728–735.

    Article  CAS  Google Scholar 

  8. Clarke MC, Savill J, Jones DB, Noble BS, Brown SB . Compartmentalized megakaryocyte death generates functional platelets committed to caspase-independent death. J Cell Biol 2003; 160: 577–587.

    Article  CAS  Google Scholar 

  9. De Botton S, Sabri S, Daugas E, Zermati Y, Guidotti JE, Hermine O et al. Platelet formation is the consequence of caspase activation within megakaryocytes. Blood 2002; 100: 1310–1317.

    Article  CAS  Google Scholar 

  10. Kaluzhny Y, Yu G, Sun S, Toselli PA, Nieswandt B, Jackson CW et al. BclxL overexpression in megakaryocytes leads to impaired platelet fragmentation. Blood 2002; 100: 1670–1678.

    Article  CAS  Google Scholar 

  11. Kozuma Y, Kojima H, Yuki S, Suzuki H, Nagasawa T . Continuous expression of Bcl-xL protein during megakaryopoiesis is post-translationally regulated by thrombopoietin-mediated Akt activation, which prevents the cleavage of Bcl-xL. J Thromb Haemost 2007; 5: 1274–1282.

    Article  CAS  Google Scholar 

  12. Lamkanfi M, Festjens N, Declercq W, Vanden Berghe T, Vandenabeele P . Caspases in cell survival, proliferation and differentiation. Cell Death Differ 2007; 14: 44–55.

    Article  CAS  Google Scholar 

  13. Carlile GW, Smith DH, Wiedmann M . Caspase-3 has a nonapoptotic function in erythroid maturation. Blood 2004; 103: 4310–4316.

    Article  CAS  Google Scholar 

  14. Zermati Y, Garrido C, Amsellem S, Fishelson S, Bouscary D, Valensi F et al. Caspase activation is required for terminal erythroid differentiation. J Exp Med 2001; 193: 247–254.

    Article  CAS  Google Scholar 

  15. Ogilvy S, Metcalf D, Print CG, Bath ML, Harris AW, Adams JM . Constitutive Bcl-2 expression throughout the hematopoietic compartment affects multiple lineages and enhances progenitor cell survival. Proc Natl Acad Sci USA 1999; 96: 14943–14948.

    Article  CAS  Google Scholar 

  16. Pronk CJ, Rossi DJ, Månsson R, Attema JL, Norddahl GL, Chan CK et al. Elucidation of the phenotypic, functional, and molecular topography of a myeloerythroid progenitor cell hierarchy. Cell Stem Cell 2007; 1: 428–442.

    Article  CAS  Google Scholar 

  17. Kojima H, Moroi M, Jung SM, Goto S, Tamura N, Kozuma Y et al. Characterization of a patient with glycoprotein (GP) VI deficiency possessing neither anti-GPVI autoantibody nor genetic aberration. J Thromb Haemost 2006; 4: 2433–2442.

    Article  CAS  Google Scholar 

  18. Zinkel S, Gross A, Yang E . BCL2 family in DNA damage and cell cycle control. Cell Death Differ 2006; 13: 1351–1359.

    Article  CAS  Google Scholar 

  19. Sordet O, Rébé C, Plenchette S, Zermati Y, Hermine O, Vainchenker W et al. Specific involvement of caspases in the differentiation of monocytes into macrophages. Blood 2002; 100: 4446–4453.

    Article  CAS  Google Scholar 

  20. Kolbus A, Pilat S, Husak Z, Deiner EM, Stengl G, Beug H et al. Raf-1 antagonizes erythroid differentiation by restraining caspase activation. J Exp Med 2002; 196: 1347–1353.

    Article  CAS  Google Scholar 

  21. Launay S, Hermine O, Fontenay M, Kroemer G, Solary E, Garrido C . Vital functions for lethal caspases. Oncogene 2005; 24: 5137–5148.

    Article  CAS  Google Scholar 

  22. Fernando P, Kelly JF, Balazsi K, Slack RS, Megeney LA . Caspase 3 activity is required for skeletal muscle differentiation. Proc Natl Aca Sci USA 2002; 99: 11025–11030.

    Article  CAS  Google Scholar 

  23. Sun S, Ravid K . Role of a serine/threonine kinase, Mst1, in megakaryocyte differentiation. J Cell Biochem 1999; 76: 44–60.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported in part by a grant from the Japanese Ministry of Education, Culture, Sports, Science and Technology (HK), and Mitsubishi Pharma Research Foundation (HK).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to H Kojima.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kozuma, Y., Yuki, S., Ninomiya, H. et al. Caspase activation is involved in early megakaryocyte differentiation but not in platelet production from megakaryocytes. Leukemia 23, 1080–1086 (2009). https://doi.org/10.1038/leu.2009.7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/leu.2009.7

Keywords

This article is cited by

Search

Quick links