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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
Pt 19
pubmed:dateCreated
2003-9-3
pubmed:abstractText
The role of cytoskeletal elements in regulating transport and docking steps that precede exocytosis of secretory organelles is not well understood. We have used Total Internal Reflection Fluorescence (TIRF) microscopy to visualize the three-dimensional motions of secretory organelles near the plasma membrane in living endothelial cells. Weibel-Palade bodies (WPb), the large tubular storage organelles for von Willebrand factor, were labelled with Rab27a-GFP. By contrast, green fluorescent protein (GFP)-tagged tissue-type plasminogen activator (tPA-GFP) labelled submicron vesicular organelles. Both populations of GFP-labelled organelles underwent stimulated exocytosis. The movement of these morphologically distinct organelles was measured within the evanescent field that penetrated the first 200 nm above the plasma membrane. WPb and tPA-GFP vesicles displayed long-range bidirectional motions and short-range diffusive-like motions. Rotating and oscillating WPb were also observed. TIRF microscopy enabled us to quantify the contribution of actin and microtubules and their associated motors to the organelle motions close to the plasma membrane. Long-range motions, as well as WPb rotations and oscillations, were microtubule-and kinesin-dependent. Disruption of the actin cytoskeleton and inhibition of myosin motors increased the number of long-range motions and, in the case of WPb, their velocity. The actin and microtubules had opposite effects on the mobility of organelles undergoing short-range motions. Actin reduced the mobility and range of motion of both WPb and tPA vesicles, whereas microtubules and kinesin motors increased the mobility of WPb. The results show that the dynamics of endothelial secretory organelles close to the plasma membrane are controlled by the opposing roles of the microtubule and actin cytoskeletal transport systems.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0021-9533
pubmed:author
pubmed:issnType
Print
pubmed:day
1
pubmed:volume
116
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
3927-38
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:12928328-Actins, pubmed-meshheading:12928328-Cell Membrane, pubmed-meshheading:12928328-Cells, Cultured, pubmed-meshheading:12928328-Cytoskeleton, pubmed-meshheading:12928328-Endothelial Cells, pubmed-meshheading:12928328-Exocytosis, pubmed-meshheading:12928328-Green Fluorescent Proteins, pubmed-meshheading:12928328-Humans, pubmed-meshheading:12928328-Kinesin, pubmed-meshheading:12928328-Luminescent Proteins, pubmed-meshheading:12928328-Microscopy, Fluorescence, pubmed-meshheading:12928328-Microtubules, pubmed-meshheading:12928328-Recombinant Fusion Proteins, pubmed-meshheading:12928328-Secretory Vesicles, pubmed-meshheading:12928328-Tissue Plasminogen Activator, pubmed-meshheading:12928328-Weibel-Palade Bodies, pubmed-meshheading:12928328-rab GTP-Binding Proteins, pubmed-meshheading:12928328-von Willebrand Factor
pubmed:year
2003
pubmed:articleTitle
Interaction of the actin cytoskeleton with microtubules regulates secretory organelle movement near the plasma membrane in human endothelial cells.
pubmed:affiliation
National Institute for Medical Research, London NW7 1AA, UK. jmannev@nimr.mrc.ac.uk
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't