The actin cytoskeleton differentially regulates platelet alpha-granule and dense-granule secretion.
Stimulation of platelets with strong agonists results in centralization of cytoplasmic organelles and secretion of granules. These observations have led to the supposition that cytoskeletal contraction facilitates granule release by promoting the interaction of granules with one another and with membranes of the open canalicular system. Yet, the influence of the actin cytoskeleton in controlling the membrane fusion events that mediate granule secretion remains largely unknown. To evaluate the role of the actin cytoskeleton in platelet granule secretion, we have assessed the effects of latrunculin A and cytochalasin E on granule secretion. Exposure of platelets to low concentrations of these reagents resulted in acceleration and augmentation of agonist-induced alpha-granule secretion with comparatively modest effects on dense granule secretion. In contrast, exposure of platelets to high concentrations of latrunculin A inhibited agonist-induced alpha-granule secretion but stimulated dense granule secretion. Incubation of permeabilized platelets with low concentrations of latrunculin A primed platelets for Ca(2+)- or guanosine triphosphate (GTP)-gamma-S-induced alpha-granule secretion. Latrunculin A-dependent alpha-granule secretion was inhibited by antibodies directed at vesicle-associated membrane protein (VAMP), demonstrating that latrunculin A supports soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) protein-dependent membrane fusion. These results indicate that the actin cytoskeleton interferes with platelet exocytosis and differentially regulates alpha-granule and dense granule secretion.
Flaumenhaft
Robert
R
Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02115, USA. rflaumen@bidmc.harvard.edu
Dilks
James R
JR
Rozenvayn
Nataliya
N
Monahan-Earley
Rita A
RA
Feng
Dian
D
Dvorak
Ann M
AM
D000199:Actins / Q000378:metabolism* / Q000648:ultrastructure; D001792:Blood Platelets / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000819:agonists; D015550:Cell Degranulation / Q000187:drug effects; D003572:Cytochalasins / Q000494:pharmacology; D003594:Cytoplasmic Granules / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D003599:Cytoskeleton / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D006801:Humans; D007700:Kinetics; D046529:Microscopy, Electron, Transmission; D010446:Peptide Fragments / Q000494:pharmacology; D050600:SNARE Proteins; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013844:Thiazoles / Q000819:agonists; D053778:Thiazolidines; D033921:Vesicular Transport Proteins / Q000378:metabolism
D000199:Actins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D003572:Cytochalasins; D010446:Peptide Fragments; D050600:SNARE Proteins; D013844:Thiazoles; D053778:Thiazolidines; D033921:Vesicular Transport Proteins; C082835:thrombin receptor peptide (42-47); C007215:cytochalasin E; D013755:Tetradecanoylphorbol Acetate; C037067:latrunculin A