|Distinct functional effects for dynamin 3 during megakaryocytopoiesis.|
|PMID: 21671749 Owner: NLM Status: MEDLINE|
|Dynamin 3 (DNM3) is a member of a family of motor proteins that participate in a number of membrane rearrangements such as cytokinesis, budding of transport vesicles, phagocytosis, and cell motility. Recently, DNM3 was implicated as having a role in megakaryocyte (MK) development. To further investigate the functional role of DNM3 during megakaryocytopoiesis, we introduced sequence-specific short hairpin RNAs (shRNAs) into developing MKs. The results showed that knockdown of DNM3 inhibited a stage of MK development that involved progenitor amplification. This was evident by significant decreases in the number of colony forming unit-megakaryocytes, the total number of nucleated cells, and the number of CD41(+) and CD61(+) MKs produced in culture. Using a styrl membrane dye to quantify the demarcation membrane system (DMS) of terminally differentiated MKs, we found that DNM3 co-localized with the DMS and that DNM3 lentiviral shRNAs precluded the formation of the DMS. Knockdown of dynamin 3 in murine MKs also caused a decrease in the number of morphologically large MKs and the overall size of large MKs was decreased relative to controls. MK protein lysates were used in overlay blots to show that both DNM3 and actin bind to nonmuscle myosin IIA (MYH9). Consistent with these observations, immunofluorescence studies of MKs and proplatelet processes showed co-localization of DNM3 with MYH9. Overall, these studies demonstrate that DNM3 not only participates in MK progenitor amplification, but is also involved in cytoplasmic enlargement and the formation of the DMS.|
|Wenjing Wang; Diana M Gilligan; Sijie Sun; Xiaoping Wu; Jo-Anna Reems|
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|Type: Journal Article; Research Support, N.I.H., Extramural Date: 2011-08-04|
|Title: Stem cells and development Volume: 20 ISSN: 1557-8534 ISO Abbreviation: Stem Cells Dev. Publication Date: 2011 Dec|
|Created Date: 2011-11-29 Completed Date: 2012-03-15 Revised Date: 2013-02-19|
Medline Journal Info:
|Nlm Unique ID: 101197107 Medline TA: Stem Cells Dev Country: United States|
|Languages: eng Pagination: 2139-51 Citation Subset: IM|
|Puget Sound Blood Center, Seattle, Washington 98104, USA. firstname.lastname@example.org|
|APA/MLA Format Download EndNote Download BibTex|
Blood Platelets / drug effects, metabolism
Cell Differentiation / drug effects
Cell Proliferation / drug effects
DNA / metabolism
Down-Regulation / drug effects
Dynamin III / metabolism*
Gene Knockdown Techniques
Indoles / pharmacology
Lentivirus / genetics
Megakaryocytes / cytology, drug effects, metabolism
Molecular Sequence Data
Myosin Heavy Chains / metabolism
Protein Binding / drug effects
Protein Transport / drug effects
RNA, Small Interfering / metabolism
Stem Cells / cytology, drug effects, metabolism
Sulfonamides / pharmacology
Thrombopoiesis* / drug effects
|P50HL081015/HL/NHLBI NIH HHS|
|0/Actins; 0/Indoles; 0/Myosin Heavy Chains; 0/RNA, Small Interfering; 0/SU 6656; 0/Sulfonamides; 9007-49-2/DNA; EC 22.214.171.124/Dynamin III|
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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