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Echicetin Coated Polystyrene Beads: A Novel Tool to Investigate GPIb-Specific Platelet Activation and Aggregation

Please always quote using this URN: urn:nbn:de:bvb:20-opus-119815
  • von Willebrand factor/ristocetin (vWF/R) induces GPIb-dependent platelet agglutination and activation of αIIbβ3 integrin, which also binds vWF. These conditions make it difficult to investigate GPIb-specific signaling pathways in washed platelets. Here, we investigated the specific mechanisms of GPIb signaling using echicetin-coated polystyrene beads, which specifically activate GPIb. We compared platelet activation induced by echicetin beads to vWF/R. Human platelets were stimulated with polystyrene beads coated with increasing amounts ofvon Willebrand factor/ristocetin (vWF/R) induces GPIb-dependent platelet agglutination and activation of αIIbβ3 integrin, which also binds vWF. These conditions make it difficult to investigate GPIb-specific signaling pathways in washed platelets. Here, we investigated the specific mechanisms of GPIb signaling using echicetin-coated polystyrene beads, which specifically activate GPIb. We compared platelet activation induced by echicetin beads to vWF/R. Human platelets were stimulated with polystyrene beads coated with increasing amounts of echicetin and platelet activation by echicetin beads was then investigated to reveal GPIb specific signaling. Echicetin beads induced αIIbβ3-dependent aggregation of washed platelets, while under the same conditions vWF/R treatment led only to αIIbβ3-independent platelet agglutination. The average distance between the echicetin molecules on the polystyrene beads must be less than 7 nm for full platelet activation, while the total amount of echicetin used for activation is not critical. Echicetin beads induced strong phosphorylation of several proteins including p38, ERK and PKB. Synergistic signaling via P2Y12 and thromboxane receptor through secreted ADP and TxA2, respectively, were important for echicetin bead triggered platelet activation. Activation of PKG by the NO/sGC/cGMP pathway inhibited echicetin bead-induced platelet aggregation. Echicetin-coated beads are powerful and reliable tools to study signaling in human platelets activated solely via GPIb and GPIb-triggered pathways.show moreshow less

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Metadaten
Author: Alexey Navdaev, Hariharan Subramanian, Alexey Petunin, Kenneth J. Clemetson, Stepan Gambaryan, Ulrich Walter
URN:urn:nbn:de:bvb:20-opus-119815
Document Type:Journal article
Faculties:Medizinische Fakultät / Institut für Klinische Biochemie und Pathobiochemie
Language:English
Parent Title (English):PLoS ONE
ISSN:1932-6203
Year of Completion:2014
Volume:9
Issue:4
Pagenumber:e93569
Source:PLoS ONE 9(4): e93569. doi:10.1371/journal.pone.0093569
DOI:https://doi.org/10.1371/journal.pone.0093569
Pubmed Id:http://www.ncbi.nlm.nih.gov/pubmed?term=24705415
Dewey Decimal Classification:6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 612 Humanphysiologie
Tag:ERK signaling cascade; integrins; phosphorylation; platelet activation; platelet aggregation; platelets; polystyrene; tyrosine
Release Date:2015/11/13
Licence (German):License LogoCC BY: Creative-Commons-Lizenz: Namensnennung