Impaired platelet aggregation and sustained bleeding in mice lacking the fibrinogen motif bound by integrin alpha IIb beta 3.
Blood loss at sites of vascular rupture is controlled by the adhesion and aggregation of platelets and the formation of an insoluble fibrin matrix. Fibrinogen is considered to be critical in these processes by both providing an abundant dimeric ligand for alpha IIb beta 3-mediated platelet aggregation, and serving as the fundamental building block of the fibrin polymer. To establish an in vivo model system to examine in detail the importance of alpha IIb beta 3-fibrinogen interactions in platelet function, hemostasis, response to injury and vasoocclusive disease, and to test the prevailing hypothesis that the C-terminal segment of the fibrinogen gamma chain is essential for alpha IIb beta 3 binding, we have used gene-targeting technology in mice to eliminate the last five residues (QAGDV) from the gamma chain. Mice homozygous for the modified gamma chain gene (gamma delta 5/gamma delta 5) displayed a generally normal hematological profile, including normal platelet count, plasma fibrinogen level, clotting time and fibrin crosslinking. However, both gamma delta 5-fibrinogen binding to alpha IIb beta 3 and platelet aggregation were highly defective. Remarkably, another alpha IIb beta 3-dependent process, clot retraction, was unaffected by the gamma delta 5 mutation. Despite the preservation of clotting function, gamma delta 5/gamma delta 5 mice were unable to control blood loss following a surgical challenge and occasionally developed fatal neonatal bleeding events.
Holmbäck
K
K
Division of Developmental Biology, Children's Hospital Research Foundation, Cincinnati, OH 45229, USA.
Danton
M J
MJ
Suh
T T
TT
Daugherty
C C
CC
Degen
J L
JL
D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn; D001483:Base Sequence; D001760:Bleeding Time; D001777:Blood Coagulation / Q000235:genetics / Q000502:physiology; D003432:Cross-Linking Reagents; D004247:DNA / Q000235:genetics; D005260:Female; D005340:Fibrinogen / Q000737:chemistry / Q000235:genetics* / Q000502:physiology*; D006470:Hemorrhage / Q000235:genetics / Q000503:physiopathology; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010974:Platelet Aggregation / Q000235:genetics* / Q000502:physiology*; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000235:genetics* / Q000502:physiology*; D011487:Protein Conformation; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion
D003432:Cross-Linking Reagents; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D012333:RNA, Messenger; D005340:Fibrinogen; D004247:DNA