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PMID: 19471020 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Differential expression of NADPH oxidases in megakaryocytes and their role in polyploidy.

Blood ·Vol. 114 ·No. 6 ·2009-08-06 ·Pages 1243-9

McCrann DJ, Eliades A, Makitalo M, Matsuno K, Ravid K

Abstract

Megakaryocytes (MKs) undergo an endomitotic cell cycle, leading to polyploidy. We examined the expression of the flavoproteins and oxidative stress-promoting enzymes, NADPH oxidases (Nox's), in MKs because of their known role in promoting the cell cycle. Although the expression of Nox isoforms varies between cell types, they are induced at the mRNA level by mitogenic stimuli. Western blotting or reverse transcription-polymerase chain reaction of purified mouse MKs isolated from thrombopoietin (TPO)-treated bone marrow (BM) cultures indicated high expression of Nox1, a weak expression of Nox4, and no significant expression of Nox2. Immunofluorescence of freshly isolated MKs confirmed strong expression of Nox1 in one-third of MKs, whereas Nox1 staining was detected in nearly all MKs in TPO-stimulated BM cultures. Treatment of mouse BM cultures with Nox inhibitors resulted in accumulation of MKs with low DNA content levels and significant reduction of higher ploidy MKs. Purified, Nox-inhibited MKs showed a notable decrease in the level of the G(1) phase cyclin E, a cyclin associated with MK polyploidy, and its up-regulation restored most of the effect of Nox inhibitors. Hence, this study shows the expression of Nox isoforms in MKs and highlights a potential role of flavoproteins in promoting polyploidization in this lineage.

MeSH Terms
Animals Bone Marrow/enzymology Enzyme Inhibitors/pharmacology G1 Phase/drug effects,physiology Gene Expression Regulation, Enzymologic/drug effects,physiology Isoenzymes/biosynthesis Megakaryocytes/enzymology Membrane Glycoproteins/biosynthesis Mice Mice, Knockout NADH, NADPH Oxidoreductases/biosynthesis NADPH Oxidase 1 NADPH Oxidase 2 NADPH Oxidase 4 NADPH Oxidases/biosynthesis Oxidative Stress/drug effects,physiology Ploidies Thrombopoietin/pharmacology Tissue Culture Techniques
Chemicals
Enzyme Inhibitors Isoenzymes Membrane Glycoproteins Thrombopoietin NADH, NADPH Oxidoreductases Cybb protein, mouse NADPH Oxidase 1 NADPH Oxidase 2 NADPH Oxidase 4 NADPH Oxidases NOX1 protein, mouse Nox4 protein, mouse
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
McCrann Donald J
Department of Biochemistry, Whitaker Cardiovascular Institute, Boston University School of Medicine, MA 02118, USA.
Eliades Alexia
Makitalo Maria
Matsuno Kuniharu
Ravid Katya
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Article Info
Journal
Blood
Abbr.
Blood
ISSN
1528-0020
DOI
10.1182/blood-2008-12-195883
Published
2009-08-06
Epub
2009-00-26
Pages
1243-9
Language
English
Region
United States
NLM ID
7603509
PMCID
PMC2723018
Subset
IM
Grants
NHLBI NIH HHS · R01 HL080442 · United States
NHLBI NIH HHS · T32 HL007969 · United States
NHLBI NIH HHS · HL80442 · United States
External Links
PubMed source DOI: 10.1182/blood-2008-12-195883 PMC Full Text View journal details
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