Calyculin A retraction of mature megakaryocytes proplatelets from embryonic stem cells

Platelets are produced by megakaryocytes (MKs) through proplatelet formation (PPF), or cytoplasmic extensions, in vitro. Through the use of video-enhanced light microscopy, as well as localization of cytoskeletal proteins by confocal microscopy, the reaction of fully mature MK proplatelets, derived...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2008-02, Vol.366 (3), p.763-768
Main Authors: Tamaru, Satoshi, Kitajima, Kenji, Nakano, Tohru, Eto, Koji, Yazaki, Akira, Kobayashi, Toshihiko, Matsumoto, Takeshi, Wada, Hideo, Katayama, Naoyuki, Nishikawa, Masakatsu
Format: Article
Language:English
Subjects:
Actin
Animals
Blood Platelets - cytology
Blood Platelets - physiology
Calyculin A
Cell Culture Techniques - methods
Cell Differentiation
Cell Line
Embryonic stem cell
Embryonic Stem Cells - cytology
Embryonic Stem Cells - physiology
Megakaryocyte
Megakaryocytes - cytology
Megakaryocytes - physiology
Mice
Myosin light chain phosphorylation
OP9
Oxazoles - metabolism
Proplatelet formation
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Summary:Platelets are produced by megakaryocytes (MKs) through proplatelet formation (PPF), or cytoplasmic extensions, in vitro. Through the use of video-enhanced light microscopy, as well as localization of cytoskeletal proteins by confocal microscopy, the reaction of fully mature MK proplatelets, derived from murine embryonic stem cells, to various agents was studied. Calyculin A (protein phosphatase 1/2A inhibitor) treatment induced proplatelet retraction. In MKs with PPF, the expression of actin, myosin IIA, monophosphorylated myosin light chain (MLC-P1), and diphosphorylated myosin light chain (MLC-P2) was diffusely located. Following calyculin A treatment, actin was diffusely localized in retracted MKs and was expressed particularly in the periphery. MLC-P1 was also localized primarily in the periphery; however, MLC-P2 was expressed mostly in the inner area of proplatelets. Protein phosphatase inhibitors may result in increased hyperphosphorylation of localized MLC, which could alter the balance of actomyosin force in a cell, and therefore induce proplatelets retraction.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2007.12.032