What is vinculin needed for in platelets?
Summary.


BACKGROUND
Vinculin links integrins to the cell cytoskeleton by virtue of its binding to proteins such as talin and F-actin. It has been implicated in the transmission of mechanical forces from the extracellular matrix to the cytoskeleton of migrating cells. Vinculin's function in platelets is unknown.


OBJECTIVE
To determine whether vinculin is required for the functions of platelets and their major integrin, α(IIb) β(3) .


METHODS
The murine vinculin gene (Vcl) was deleted in the megakaryocyte/platelet lineage by breeding Vcl fl/fl mice with Pf4-Cre mice. Platelet and integrin functions were studied in vivo and ex vivo.


RESULTS
Vinculin was undetectable in platelets from Vcl fl/fl Cre(+) mice, as determined by immunoblotting and fluorescence microscopy. Vinculin-deficient megakaryocytes exhibited increased membrane tethers in response to mechanical pulling on α(IIb) β(3) with laser tweezers, suggesting that vinculin helps to maintain membrane cytoskeleton integrity. Surprisingly, vinculin-deficient platelets displayed normal agonist-induced fibrinogen binding to α(IIb) β(3) , aggregation, spreading, actin polymerization/organization, clot retraction and the ability to form a procoagulant surface. Furthermore, vinculin-deficient platelets adhered to immobilized fibrinogen or collagen normally, under both static and flow conditions. Tail bleeding times were prolonged in 59% of vinculin-deficient mice. However, these mice exhibited no spontaneous bleeding and they formed occlusive platelet thrombi comparable to those in wild-type littermates in response to carotid artery injury with FeCl(3) .


CONCLUSIONS
Despite promoting membrane cytoskeleton integrity when mechanical force is applied to α(IIb) β(3) , vinculin is not required for the traditional functions of α(IIb) β(3) or the platelet actin cytoskeleton.
Mitsios
J V
JV
Department of Medicine, University of California, San Diego, La Jolla, CA 92093-0726, USA.
Prevost
N
N
Kasirer-Friede
A
A
Gutierrez
E
E
Groisman
A
A
Abrams
C S
CS
Wang
Y
Y
Litvinov
R I
RI
Zemljic-Harpf
A
A
Ross
R S
RS
Shattil
S J
SJ
D000199:Actins / Q000737:chemistry*; D000818:Animals; D001792:Blood Platelets / Q000378:metabolism*; D019070:Cell Lineage; D003094:Collagen / Q000737:chemistry; D003599:Cytoskeleton / Q000378:metabolism*; D005340:Fibrinogen / Q000737:chemistry; D017353:Gene Deletion; D008533:Megakaryocytes / Q000166:cytology; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence / Q000379:methods; D010974:Platelet Aggregation; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000378:metabolism; D016596:Vinculin / Q000378:metabolism / Q000502:physiology*
D000199:Actins; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D016596:Vinculin; D005340:Fibrinogen; D003094:Collagen