Intracellular calcium response of endothelial cells exposed to flow in the presence of thrombin or histamine Laura M. Worthen, MS, Matthias U. Nollert,

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Presentation transcript:

Intracellular calcium response of endothelial cells exposed to flow in the presence of thrombin or histamine Laura M. Worthen, MS, Matthias U. Nollert, PhD Journal of Vascular Surgery Volume 32, Issue 3, Pages 593-601 (September 2000) DOI: 10.1067/mva.2000.106955 Copyright © 2000 Society for Vascular Surgery and The American Association for Vascular Surgery, a Chapter of the International Society for Cardiovascular Surgery Terms and Conditions

Fig. 1 Intracellular calcium response of a single 40× field of view of HUVECs (approximately 60-80 cells) exposed to flow at several concentrations of histamine. On each graph, first vertical bar represents time at which flow was started; second vertical bar represents time at which, according to prediction of convection/diffusion model, histamine concentration at wall reaches 50% of bulk concentration. Time to reach 50% of bulk value is 13 s for 0.2 dyne/cm2, 3.4 s for 1 dyne/cm2, and 0.4 s for 20 dyne/cm2. Journal of Vascular Surgery 2000 32, 593-601DOI: (10.1067/mva.2000.106955) Copyright © 2000 Society for Vascular Surgery and The American Association for Vascular Surgery, a Chapter of the International Society for Cardiovascular Surgery Terms and Conditions

Fig. 2 Maximum increase in intracellular calcium concentration for endothelial cells exposed to shear stress and either histamine or thrombin. Results from 6 to 8 different cord pools were averaged for each data point. Error bars represent SEM. A, Cells exposed to histamine. B, Cells exposed to thrombin. Asterisk indicates significant difference. Journal of Vascular Surgery 2000 32, 593-601DOI: (10.1067/mva.2000.106955) Copyright © 2000 Society for Vascular Surgery and The American Association for Vascular Surgery, a Chapter of the International Society for Cardiovascular Surgery Terms and Conditions

Fig. 3 Response of single cells in monolayer of primary HUVECs that were exposed to thrombin and shear stresses of 0.2, 1.0, or 20 dyne/cm2. Black vertical bar at approximately 20 seconds indicates time at which flow was started. Small vertical bars on each single cell tracing illustrate time required for concentration of thrombin near cell surface to reach 50% of its bulk value, as determined by solution to convection/diffusion equations; this time is 25.3 s for 0.2 dyne/cm2, 7.7 s for 1 dyne/cm2, and 0.6 s for 20 dyne/cm2. A, Response to 0.02 U/mL of thrombin. B, Response to 0.1 U/mL of thrombin. C, Response to 0.2 U/mL of thrombin. Journal of Vascular Surgery 2000 32, 593-601DOI: (10.1067/mva.2000.106955) Copyright © 2000 Society for Vascular Surgery and The American Association for Vascular Surgery, a Chapter of the International Society for Cardiovascular Surgery Terms and Conditions

Fig. 4 Confluent monolayers of endothelial cells were exposed to shear stress and thrombin. Percent of cells that showed increase of at least 20 nmol in intracellular calcium was determined for each field of view. Two to 5 experiments were used for each shear stress and concentration of thrombin. A is for 0.02 U/mL thrombin; B is for 0.2 U/mL thrombin. Journal of Vascular Surgery 2000 32, 593-601DOI: (10.1067/mva.2000.106955) Copyright © 2000 Society for Vascular Surgery and The American Association for Vascular Surgery, a Chapter of the International Society for Cardiovascular Surgery Terms and Conditions