Figure 5
Figure 5. GPVI mediates vascular repair following neutrophil-induced injury. (A) Comparison of platelet accumulation at unstimulated control sites or at the site of the rpA reaction in the skin of control wild-type, thrombocytopenic, and GPVI−/− mice, as assessed by measurement of skin PF4 content. n = 6-15 mice per group. # indicates a significant difference (P < .0001) as compared with the PF4 content in unstimulated control skin of wild-type mice. (B) Platelets and neutrophils were stained in vivo using fluorescent antibodies to GPIbβ and Ly-6G, respectively, and their interactions with vessels during the rpA reaction were analyzed by intravital microscopy through a dorsal skinfold chamber. Whether at the capillary, postcapillary venule, or vein level, single platelets were seen binding directly to the vessel lining (white arrowheads) or to adherent neutrophils (white asterisks). Vessel edges are highlighted by dotted lines. The images shown were taken between 90 minutes and 2 hours after inducing the rpA reaction and are representative of 4 independent experiments. Scale bar, 100 µm. (C) The interactions of fluorescent microspheres (2 µm diameter) coated with a chimeric form of GPVI (GPVI-Fc) with the inflamed vasculature were analyzed by intravital microscopy. GPVI-Fc-coated microspheres accumulated specifically at sites of leukocyte recruitment and infiltration (white asterisks) identified by rhodamine-6G labeling. Observation of the GPVI-Fc–coated microspheres at higher magnification revealed individual microspheres binding directly to the vessel wall (black arrowhead) or to adherent leukocytes (white arrowhead). Vessel edges are highlighted in white. Scale bars, 200 µm (left) and 50 µm (right). The images shown are representative of 4 independent experiments. (D) Quantification of control and GPVI-Fc–coated microsphere accumulation at unstimulated control sites or at the site of the rpA reaction in the skin of wild-type mice that were immunodepleted or not for neutrophils, as indicated. n = 5-8 mice per group. NS, not significant.

GPVI mediates vascular repair following neutrophil-induced injury. (A) Comparison of platelet accumulation at unstimulated control sites or at the site of the rpA reaction in the skin of control wild-type, thrombocytopenic, and GPVI−/− mice, as assessed by measurement of skin PF4 content. n = 6-15 mice per group. # indicates a significant difference (P < .0001) as compared with the PF4 content in unstimulated control skin of wild-type mice. (B) Platelets and neutrophils were stained in vivo using fluorescent antibodies to GPIbβ and Ly-6G, respectively, and their interactions with vessels during the rpA reaction were analyzed by intravital microscopy through a dorsal skinfold chamber. Whether at the capillary, postcapillary venule, or vein level, single platelets were seen binding directly to the vessel lining (white arrowheads) or to adherent neutrophils (white asterisks). Vessel edges are highlighted by dotted lines. The images shown were taken between 90 minutes and 2 hours after inducing the rpA reaction and are representative of 4 independent experiments. Scale bar, 100 µm. (C) The interactions of fluorescent microspheres (2 µm diameter) coated with a chimeric form of GPVI (GPVI-Fc) with the inflamed vasculature were analyzed by intravital microscopy. GPVI-Fc-coated microspheres accumulated specifically at sites of leukocyte recruitment and infiltration (white asterisks) identified by rhodamine-6G labeling. Observation of the GPVI-Fc–coated microspheres at higher magnification revealed individual microspheres binding directly to the vessel wall (black arrowhead) or to adherent leukocytes (white arrowhead). Vessel edges are highlighted in white. Scale bars, 200 µm (left) and 50 µm (right). The images shown are representative of 4 independent experiments. (D) Quantification of control and GPVI-Fc–coated microsphere accumulation at unstimulated control sites or at the site of the rpA reaction in the skin of wild-type mice that were immunodepleted or not for neutrophils, as indicated. n = 5-8 mice per group. NS, not significant.

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