NDLI: Differential roles of microtubule assembly and sliding in proplatelet formation by megakaryocytes

Content Provider	PubMed Central
Author	Patel, Sunita R. Richardson, Jennifer L. Schulze, Harald Kahle, Eden Niels, Galjart Drabek, Ksenija Shivdasani, Ramesh A. Hartwig, John H. Italiano, Joseph E.
Copyright Year	2005
Abstract	Megakaryocytes are terminally differentiated cells that, in their final hours, convert their cytoplasm into long, branched proplatelets, which remodel into blood platelets. Proplatelets elongate at an average rate of 0.85 μm/min in a microtubule-dependent process. Addition of rhodamine-tubulin to permeabilized proplatelets, immunofluorescence microscopy of the microtubule plus-end marker end-binding protein 3 (EB3), and fluorescence time-lapse microscopy of EB3–green fluorescent protein (GFP)–expressing megakaryocytes reveal that microtubules, organized as bipolar arrays, continuously polymerize throughout the proplatelet. In immature megakaryocytes lacking proplatelets, microtubule plus-ends initiate and grow by centrosomal nucleation at rates of 8.9 to 12.3 μm/min. In contrast, plus-end growth rates of microtubules within proplatelets are highly variable (1.5-23.5 μm/min) and are both slower and faster than those seen in immature cells. Despite the continuous assembly of microtubules, proplatelets continue to elongate when net microtubule assembly is arrested. One alternative mechanism for force generation is microtubule sliding. Triton X-100–permeabilized proplatelets containing dynein and its regulatory complex, dynactin, but not kinesin, elongate with the addition of adenosine triphosphate (ATP) at a rate of 0.65 μm/min. Retroviral expression in megakaryocytes of dynamitin (p50), which disrupts dynactindynein function, inhibits proplatelet elongation. We conclude that while continuous polymerization of microtubules is necessary to support the enlarging proplatelet mass, the sliding of overlapping microtubules is a vital component of proplatelet elongation.
Related Links	http://dx.doi.org/10.1182/blood-2005-06-2204
Ending Page	4085
Page Count	10
Starting Page	4076
File Format	PDF
ISSN	00064971
e-ISSN	15280020
Journal	Blood
Issue Number	13
Volume Number	106
Language	English
Publisher	© 2005 by The American Society of Hematology
Publisher Date	2005-12-15
Access Restriction	Open
Rights Holder	© 2005 by The American Society of Hematology
Subject Keyword	Immunology Cell Biology Biochemistry Hematology Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Cell Biology Hematology Biochemistry Immunology

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