MyosinIIa contractility is required for maintenance of platelet structure during spreading on collagen and contributes to thrombus stability.
BACKGROUND
MyosinIIs are adenosine triphosphate-driven molecular motors that form part of a cell's contractile machinery. They are activated by phosphorylation of their light chains, by either activation of myosin light chain (MLC) kinase or inhibition of MLC phosphatase via Rho kinase (ROCK). MyosinIIa phosphorylation underlies platelet rounding and stress fiber formation.


OBJECTIVE
To identify the functional significance of myosinIIa in platelet spreading and thrombus formation on collagen using inhibitors of ROCK (Y27632) and myosinII (blebbistatin).


RESULTS
Stress fiber formation on collagen is inhibited by both Y27632 and blebbistatin. A substantial proportion of spread platelets generate internal holes or splits on collagen, presumably because of a reduction in contractile strength. Platelet integrity, however, is maintained. In an in vitro model, thrombus embolization on collagen is increased in the presence of Y27632 and blebbistatin at intermediate shear, leading to a reduction in platelet aggregate growth. Moreover, Y27632 causes a marked reduction in thrombus formation in an in vivo laser-injury model.


CONCLUSIONS
MyosinIIa contractility is required for maintenance of platelet structure during spreading on collagen and contributes to thrombus stability.
Calaminus
S D J
SD
Centre for Cardiovascular Sciences, Institute of Biomedical Research, The Medical School, University of Birmingham, Birmingham, UK. sdc379@bham.ac.uk
Auger
J M
JM
McCarty
O J T
OJ
Wakelam
M J O
MJ
Machesky
L M
LM
Watson
S P
SP
D000199:Actins / Q000378:metabolism; D000577:Amides / Q000494:pharmacology; D001792:Blood Platelets / Q000166:cytology / Q000378:metabolism; D002465:Cell Movement; D003094:Collagen / Q000737:chemistry / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D006576:Heterocyclic Compounds, 4 or More Rings / Q000494:pharmacology; D006801:Humans; D008954:Models, Biological; D018994:Myosin Light Chains / Q000737:chemistry; D024507:Nonmuscle Myosin Type IIA / Q000096:biosynthesis* / Q000502:physiology*; D010766:Phosphorylation; D010974:Platelet Aggregation; D011725:Pyridines / Q000494:pharmacology; D013927:Thrombosis / Q000378:metabolism; D054460:rho-Associated Kinases / Q000378:metabolism
D000199:Actins; D000577:Amides; D006576:Heterocyclic Compounds, 4 or More Rings; D018994:Myosin Light Chains; D011725:Pyridines; C108830:Y 27632; C472645:blebbistatin; D003094:Collagen; D054460:rho-Associated Kinases; D024507:Nonmuscle Myosin Type IIA