title: Hierarchical organization of the hemostatic response to penetrating injuries in the mouse macrovasculature.
abstract: Essentials Methods were developed to image the hemostatic response in mouse femoral arteries in real time. Penetrating injuries produced thrombi consisting primarily of platelets. Similar to arterioles, a core-shell architecture of platelet activation occurs in the femoral artery. Differences from arterioles included slower platelet activation and reduced thrombin dependence.
  
  
  
  Background Intravital studies performed in the mouse microcirculation show that hemostatic thrombi formed after penetrating injuries develop a characteristic architecture in which a core of fully activated, densely packed platelets is overlaid with a shell of less activated platelets. Objective Large differences in hemodynamics and vessel wall biology distinguish arteries from arterioles. Here we asked whether these differences affect the hemostatic response and alter the impact of anticoagulants and antiplatelet agents. Methods Approaches previously developed for intravital imaging in the mouse microcirculation were adapted to the femoral artery, enabling real-time fluorescence imaging despite the markedly thicker vessel wall. Results Arterial thrombi initiated by penetrating injuries developed the core-and-shell architecture previously observed in the microcirculation. However, although platelet accumulation was greater in arterial thrombi, the kinetics of platelet activation were slower. Inhibiting platelet ADP P2Y12receptors destabilized the shell and reduced thrombus size without affecting the core. Inhibiting thrombin with hirudin suppressed fibrin accumulation, but had little impact on thrombus size. Removing the platelet collagen receptor, glycoprotein VI, had no effect. Conclusions These results (i) demonstrate the feasibility of performing high-speed fluorescence imaging in larger vessels and (ii) highlight differences as well as similarities in the hemostatic response in the macro- and microcirculation. Similarities include the overall core-and-shell architecture. Differences include the slower kinetics of platelet activation and a smaller contribution from thrombin, which may be due in part to the greater thickness of the arterial wall and the correspondingly greater separation of tissue factor from the vessel lumen.
authors.affiliation: Departments of Medicine and Pharmacology, University of Pennsylvania, Philadelphia, PA, USA.|Department of Chemical and Biomolecular Engineering, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: J D
authors.initials: JD
authors.lastname: Welsh
authors.affiliation: Departments of Medicine and Pharmacology, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: I
authors.initials: I
authors.lastname: Poventud-Fuentes
authors.affiliation: Departments of Medicine and Pharmacology, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: S
authors.initials: S
authors.lastname: Sampietro
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: S L
authors.initials: SL
authors.lastname: Diamond
authors.affiliation: Departments of Medicine and Pharmacology, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: T J
authors.initials: TJ
authors.lastname: Stalker
authors.affiliation: Departments of Medicine and Pharmacology, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: L F
authors.initials: LF
authors.lastname: Brass
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology; D001160:Arterioles / Q000378:metabolism; D001777:Blood Coagulation / Q000187:drug effects; D001792:Blood Platelets / Q000378:metabolism; D005263:Femoral Artery / Q000000981:diagnostic imaging* / Q000293:injuries; D005337:Fibrin / Q000378:metabolism; D006439:Hemodynamics; D006487:Hemostasis; D000069416:Intravital Microscopy; D051379:Mice; D008810:Mice, Inbred C57BL; D008833:Microcirculation; D015539:Platelet Activation; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology; D015398:Signal Transduction; D013917:Thrombin / Q000037:antagonists & inhibitors / Q000378:metabolism; D013925:Thromboplastin / Q000378:metabolism; D013927:Thrombosis / Q000000981:diagnostic imaging / Q000188:drug therapy; D014950:Wounds, Penetrating / Q000628:therapy*
chemical_list: D000925:Anticoagulants; D010975:Platelet Aggregation Inhibitors; D000244:Adenosine Diphosphate; D005337:Fibrin; D013925:Thromboplastin; D013917:Thrombin
keywords: fibrin; platelet activation; platelet aggregation inhibitors; thrombin; thromboplastin