Enucleation of human erythroblasts involves non-muscle myosin IIB

Blood. 2012 Jan 26;119(4):1036-44. doi: 10.1182/blood-2011-06-361907. Epub 2011 Nov 2.

Abstract

Mammalian erythroblasts undergo enucleation, a process thought to be similar to cytokinesis. Although an assemblage of actin, non-muscle myosin II, and several other proteins is crucial for proper cytokinesis, the role of non-muscle myosin II in enucleation remains unclear. In this study, we investigated the effect of various cell-division inhibitors on cytokinesis and enucleation. For this purpose, we used human colony-forming unit-erythroid (CFU-E) and mature erythroblasts generated from purified CD34(+) cells as target cells for cytokinesis and enucleation assay, respectively. Here we show that the inhibition of myosin by blebbistatin, an inhibitor of non-muscle myosin II ATPase, blocks both cell division and enucleation, which suggests that non-muscle myosin II plays an essential role not only in cytokinesis but also in enucleation. When the function of non-muscle myosin heavy chain (NMHC) IIA or IIB was inhibited by an exogenous expression of myosin rod fragment, myosin IIA or IIB, each rod fragment blocked the proliferation of CFU-E but only the rod fragment for IIB inhibited the enucleation of mature erythroblasts. These data indicate that NMHC IIB among the isoforms is involved in the enucleation of human erythroblasts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amides / pharmacology
  • Aminoquinolines / pharmacology
  • Cells, Cultured
  • Cytokinesis / drug effects
  • Enzyme Inhibitors / pharmacology
  • Erythroblasts / cytology*
  • Erythroblasts / drug effects
  • Erythroblasts / metabolism*
  • Erythroid Precursor Cells / cytology
  • Erythroid Precursor Cells / drug effects
  • Erythroid Precursor Cells / metabolism
  • Erythropoiesis* / drug effects
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Heterocyclic Compounds, 4 or More Rings / pharmacology
  • Humans
  • Microfilament Proteins / antagonists & inhibitors
  • Myosins / antagonists & inhibitors
  • Nonmuscle Myosin Type IIA / antagonists & inhibitors
  • Nonmuscle Myosin Type IIA / genetics
  • Nonmuscle Myosin Type IIA / metabolism
  • Nonmuscle Myosin Type IIB / antagonists & inhibitors
  • Nonmuscle Myosin Type IIB / genetics
  • Nonmuscle Myosin Type IIB / metabolism*
  • Peptide Fragments / genetics
  • Peptide Fragments / metabolism
  • Protein Isoforms / antagonists & inhibitors
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • Pyridines / pharmacology
  • Pyrimidines / pharmacology
  • Recombinant Fusion Proteins / metabolism
  • rac1 GTP-Binding Protein / antagonists & inhibitors
  • rho-Associated Kinases / antagonists & inhibitors

Substances

  • Amides
  • Aminoquinolines
  • Enzyme Inhibitors
  • Heterocyclic Compounds, 4 or More Rings
  • Microfilament Proteins
  • NSC 23766
  • Peptide Fragments
  • Protein Isoforms
  • Pyridines
  • Pyrimidines
  • Recombinant Fusion Proteins
  • enhanced green fluorescent protein
  • Y 27632
  • Green Fluorescent Proteins
  • blebbistatin
  • rho-Associated Kinases
  • Nonmuscle Myosin Type IIA
  • Nonmuscle Myosin Type IIB
  • Myosins
  • rac1 GTP-Binding Protein