Skip to main content
SpringerLink
Log in

Internuclear chromosome distribution of dysplastic megakaryocytes in myelodysplastic syndromes is dependent on the level of ploidy

  • Research Article
  • Published:
Chromosoma Aims and scope Submit manuscript

Abstract

Megakaryopoiesis is largely disturbed in myelodysplastic syndromes (MDS), and megakaryocytes (MKs) frequently show multinucleation. Here, we investigated dysplastic mono-, bi-, and multinuclear MKs (n = 169) of seven patients with MDS and one patient with myelodysplastic/myeloproliferative neoplasm by sequential multilocus FISH. Analysis of binuclear MKs with a combined DNA content of 4 N (n = 46) indicated a significantly even (symmetric) chromosome distribution between the two separate nuclei (p = 0.0223), which suggests bipolar spindle orientation and symmetric chromosome segregation during the first endomitotic cell cycle. In contrast, multinuclear MKs of higher ploidy (>4 N, n = 108) demonstrated a significantly uneven (asymmetric) chromosome distribution between the separate nuclei (p = 0.0248). Thus, the internuclear chromosomal distribution of dysplastic MKs depends on the level of ploidy. In addition, centrosomal aberrations were not found in dysplastic MKs. Our results indicate that megakaryocytic multinucleation in MDS originates from dysregulated endomitosis, including restoration of karyokinesis.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Bartova E, Kozubek S, Jirsova P, Kozubek M, Lukasova E, Skalnikova M, Cafourkova A, Koutna I, Pasekova R (2001) Higher-order chromatin structure of human granulocytes. Chromosoma 110:360–370

    Article  PubMed  CAS  Google Scholar 

  • Braun T, Carvalho G, Grosjean J, Ades L, Fabre C, Boehrer S, Debili N, Fenaux P, Kroemer G (2007) Differentiating megakaryocytes in myelodysplastic syndromes succumb to mitochondrial derangement without caspase activation. Apoptosis 12:1101–1108

    Article  PubMed  CAS  Google Scholar 

  • Essers J, van Cappellen WA, Theil AF, van Drunen E, Jaspers NG, Hoeijmakers JH, Wyman C, Vermeulen W, Kanaar R (2005) Dynamics of relative chromosome position during the cell cycle. Mol Biol Cell 16:769–775

    Article  PubMed  CAS  Google Scholar 

  • Ganem NJ, Godinho SA, Pellman D (2009) A mechanism linking extra centrosomes to chromosomal instability. Nature 460:278–282

    Article  PubMed  CAS  Google Scholar 

  • Geddis AE, Kaushansky K (2006) Endomitotic megakaryocytes form a midzone in anaphase but have a deficiency in cleavage furrow formation. Cell Cycle 5:538–545

    Article  PubMed  CAS  Google Scholar 

  • Geddis AE, Fox NE, Tkachenko E, Kaushansky K (2007) Endomitotic megakaryocytes that form a bipolar spindle exhibit cleavage furrow ingression followed by furrow regression. Cell Cycle 6:455–460

    Article  PubMed  CAS  Google Scholar 

  • Giehl M, Fabarius A, Frank O, Hochhaus A, Hafner M, Hehlmann R, Seifarth W (2005) Centrosome aberrations in chronic myeloid leukemia correlate with stage of disease and chromosomal instability. Leukemia 19:1192–1197

    Article  PubMed  CAS  Google Scholar 

  • Hancock V, Martin JF, Lelchuk R (1993) The relationship between human megakaryocyte nuclear DNA content and gene expression. Br J Haematol 85:692–697

    Article  PubMed  CAS  Google Scholar 

  • Knockleby J, Lee H (2010) Same partners, different dance: involvement of DNA replication proteins in centrosome regulation. Cell Cycle 9:4487–4491

    Article  PubMed  CAS  Google Scholar 

  • Kwon M, Godinho SA, Chandhok NS, Ganem NJ, Azioune A, Thery M, Pellman D (2008) Mechanisms to suppress multipolar divisions in cancer cells with extra centrosomes. Genes Dev 22:2189–2203

    Article  PubMed  CAS  Google Scholar 

  • Lordier L, Jalil A, Aurade F, Larbret F, Larghero J, Debili N, Vainchenker W, Chang Y (2008) Megakaryocyte endomitosis is a failure of late cytokinesis related to defects in the contractile ring and Rho/Rock signaling. Blood 112:3164–3174

    Article  PubMed  CAS  Google Scholar 

  • Lukasova E, Kozubek S, Falk M, Kozubek M, Zaloudik J, Vagunda V, Pavlovsky Z (2004) Topography of genetic loci in the nuclei of cells of colorectal carcinoma and adjacent tissue of colonic epithelium. Chromosoma 112:221–230

    Article  PubMed  Google Scholar 

  • Mhawech P, Saleem A (2001) Myelodysplastic syndrome: review of the cytogenetic and molecular data. Crit Rev Oncol Hematol 40:229–238

    Article  PubMed  CAS  Google Scholar 

  • Minn IL, Rolls MM, Hanna-Rose W, Malone CJ (2009) SUN-1 and ZYG-12, mediators of centrosome-nucleus attachment, are a functional SUN/KASH pair in Caenorhabditis elegans. Mol Biol Cell 20:4586–4595

    Article  PubMed  CAS  Google Scholar 

  • Neben K, Giesecke C, Schweizer S, Ho AD, Kramer A (2003) Centrosome aberrations in acute myeloid leukemia are correlated with cytogenetic risk profile. Blood 101:289–291

    Article  PubMed  CAS  Google Scholar 

  • Neukirchen J, Blum S, Kuendgen A et al (2009) Platelet counts and haemorrhagic diathesis in patients with myelodysplastic syndromes. Eur J Haematol 83:477–482

    Article  PubMed  Google Scholar 

  • Nickenig C, Lang NK, Schoch C, Hiddemann W, Haferlach T (2001) New insights into the biology of multiple myeloma using a combination of May-Grunwald-Giemsa staining and fluorescence in situ hybridization techniques at the single cell level. Ann Hematol 80:662–668

    Article  PubMed  CAS  Google Scholar 

  • Papadantonakis N, Makitalo M, McCrann DJ, Liu K, Nguyen HG, Martin G, Patel-Hett S, Italiano JE, Ravid K (2008) Direct visualization of the endomitotic cell cycle in living megakaryocytes: differential patterns in low and high ploidy cells. Cell Cycle 7:2352–2356

    PubMed  CAS  Google Scholar 

  • Patel H, Gordon MY (2009) Abnormal centrosome-centriole cycle in chronic myeloid leukaemia? Br J Haematol 146:408–417

    Article  PubMed  CAS  Google Scholar 

  • Raslova H, Roy L, Vourc'h C, Le Couedic JP, Brison O, Metivier D, Feunteun J, Kroemer G, Debili N, Vainchenker W (2003) Megakaryocyte polyploidization is associated with a functional gene amplification. Blood 101:541–544

    Article  PubMed  CAS  Google Scholar 

  • Raslova H, Kauffmann A, Sekkai D, Ripoche H, Larbret F, Robert T, Le Roux DT, Kroemer G, Debili N, Dessen P, Lazar V, Vainchenker W (2007) Interrelation between polyploidization and megakaryocyte differentiation: a gene profiling approach. Blood 109:3225–3234

    Article  PubMed  CAS  Google Scholar 

  • Ravid K, Lu J, Zimmet JM, Jones MR (2002) Roads to polyploidy: the megakaryocyte example. J Cell Physiol 190:7–20

    Article  PubMed  CAS  Google Scholar 

  • Rigby PW, Dieckmann M, Rhodes C, Berg P (1977) Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251

    Article  PubMed  CAS  Google Scholar 

  • Ring D, Hubble R, Kirschner M (1982) Mitosis in a cell with multiple centrioles. J Cell Biol 94:549–556

    Article  PubMed  CAS  Google Scholar 

  • Roy L, Coullin P, Vitrat N, Hellio R, Debili N, Weinstein J, Bernheim A, Vainchenker W (2001) Asymmetrical segregation of chromosomes with a normal metaphase/anaphase checkpoint in polyploid megakaryocytes. Blood 97:2238–2247

    Google Scholar 

  • Schaefer HE, Lubbert M (2005) The hematopathological basis for studying effects of the demethylating agent 5-aza-2′-deoxycytidine (decitabine) in myelodysplasia. Ann Hematol 84(Suppl 1):67–79

    Article  PubMed  CAS  Google Scholar 

  • Schneider M, Lu W, Neumann S et al (2010) Molecular mechanisms of centrosome and cytoskeleton anchorage at the nuclear envelope. Cell Mol Life Sci (in press)

  • Shaffer LG, Tommerup N (eds) (2005) ISCN (2005): An international system for human cytogenetic nomenclature

  • Swerdlow SH, Campo E, Harris NL, Jaffe ES, Pileri SH, Stein H, Thiele J, Vardiman JW (eds) (2008) World Health Organization Classification of Tumours of haematopoietic and lymphoid tissues. WHO Press, Geneve

    Google Scholar 

  • Tefferi A, Vardiman JW (2009) Myelodysplastic syndromes. N Engl J Med 361:1872–1885

    Article  PubMed  CAS  Google Scholar 

  • van Lom K, Houtsmuller AB, van Putten WL, Slater RM, Lowenberg B (1999) Cytogenetic clonality analysis of megakaryocytes in myelodysplastic syndrome by dual-color fluorescence in situ hybridization and confocal laser scanning microscopy. Gene Chromosome Canc 25:332–338

    Article  Google Scholar 

  • Walch A, Bink K, Hutzler P, Bowering K, Letsiou I, Zitzelsberger H, Braselmann H, Stein H, Hofler H, Werner M (2001) Sequential multilocus fluorescence in situ hybridization can detect complex patterns of increased gene dosage at the single cell level in tissue sections. Lab Invest 81:1457–1459

    PubMed  CAS  Google Scholar 

  • Yan J, Zhang XX, Fetni R, Drouin R (2001) Trisomy 8 and monosomy 7 detected in bone marrow using primed in situ labeling, fluorescence in situ hybridization, and conventional cytogenetic analyses. A study of 54 cases with hematological disorders. Cancer Genet Cytogenet 125:30–40

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We are grateful to Mrs. Inga Sandrock for participating in the study as part of a laboratory internship. FISH probes were generously provided by Dr. M. Rocchi (Bari, Italy). This study was awarded a poster prize at the international meeting “Signal Transduction and Disease 2009” organized by the German, Netherlands, and Belgian Societies of Biochemistry and Molecular Biology. S.L. and M.W. further acknowledge the long-term support by the Mushett Family Foundation (Chester, NJ, US).

Author information

Authors and Affiliations

  1. Institute of Pathology, University Medical Center Freiburg, Breisacher Str. 115a, 79106, Freiburg, Germany

    Claudia Münch, Annette M. May, Jasmine Roth, Silke Lassmann & Martin Werner

  2. Institute of Medical Biometry and Medical Informatics, University Medical Center Freiburg, Stefan-Meier Str. 26, 79104, Freiburg, Germany

    Dieter Hauschke

Authors
  1. Claudia Münch
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Annette M. May
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Dieter Hauschke
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jasmine Roth
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Silke Lassmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Martin Werner
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Martin Werner.

Additional information

Communicated by F. Uhlmann

Claudia Münch and Annette M. May contributed equally to this study.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary Table 1

Results of centrosome analysis of MKs in control bone marrow smears (DOC 29 kb)

Supplementary Figure 1

Centrosome staining of control MKs. Centrosomes were stained with a monoclonal mouse antibody to pericentrin and visualized with AlexaFluor568 goat anti-mouse (red). (A) Control MK with separate centrosomes with cytoplasmic localization. (B) Control MK with centrosome cluster and additional separate centrosomes. (C) Control MK with a centrosome cluster.

High Resolution Image File (TIFF 960 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Münch, C., May, A.M., Hauschke, D. et al. Internuclear chromosome distribution of dysplastic megakaryocytes in myelodysplastic syndromes is dependent on the level of ploidy. Chromosoma 120, 265–273 (2011). https://doi.org/10.1007/s00412-011-0309-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00412-011-0309-x

Keywords

Search

Navigation