Jason G. Umans, Debra Salvi, Patrick T. Murray, Mark E. Wylam

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Selectivity of endotoxin-induced defect in endothelial calcium mobilization Jason G. Umans, Debra Salvi, Patrick T. Murray, Mark E. Wylam Kidney International Volume 54, Issue 4, Pages 1063-1069 (October 1998) DOI: 10.1046/j.1523-1755.1998.00090.x Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 1 Sample traces of endothelial [Ca2+]i versus time following stimulation with 25 μmbradykinin (BK). Preincubation with 1 μg/ml lipopolysaccharide (LPS) decreased peak response when compared with vehicle-treated control. Kidney International 1998 54, 1063-1069DOI: (10.1046/j.1523-1755.1998.00090.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 2 Dose-response and time-action data for effect of lipopolysaccharide (LPS) on subsequent bradykinin (BK)-induced Ca2+ responses (N = 12/dose and time). (A) No effect of one hour LPS exposure. (B) Significant decrease in BK response with two hours exposure to 1 μg/ml LPS. LPS incubation for four hours (C) led to dose-dependent (1pg to 1 μg/ml) decrements of BK response. Graded effects of LPS (1ng to 1 μg/ml) were also observed at six hours (D). Symbols are: (▪) control; (•) 1pg/ml; (▴) 1ng/ml; (▾) 1 μg/ml. Kidney International 1998 54, 1063-1069DOI: (10.1046/j.1523-1755.1998.00090.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 3 Lipopolysaccharide (LPS; 1 μg/ml, 18hr incubation, N = 4 to 6/dose and treatment group) decreased bradykinin (BK)-induced Ca2+ mobilization in confluent endothelial cells (A and B), whether cultured with 20% FBS (A) or previously quiesced with 1% FBS (B). By contrast, the effect of LPS was significantly attenuated in subconfluent cells (C and D), even when quiesced with 1% FBS. Symbols are: (▪) control; (•) 1pg/ml; (▴) 1ng/ml; (▾) 1 μg/ml. Kidney International 1998 54, 1063-1069DOI: (10.1046/j.1523-1755.1998.00090.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 4 Time course of endothelial [Ca2+]i following graded doses of LPS (1pg to 1mg/ml, N = 4 to 16/dose). Low doses (1pg to 1ng/ml) failed to increase Ca2+ while higher doses (1 to 10 μg/ml) led to only delayed increments of Ca2+. Only the highest doses of LPS (0.1 to 1mg/ml) increased endothelial Ca2+ within the first minute following drug addition. Symbols are: (□) control; (○) 1pg/ml LPS; (▵) 1ng/ml LPS; (▿) 1 μg/ml LPS; (X) 100 μg/ml; (*) 1mg/ml LPS. Kidney International 1998 54, 1063-1069DOI: (10.1046/j.1523-1755.1998.00090.x) Copyright © 1998 International Society of Nephrology Terms and Conditions

Figure 5 Histograms showing decreased bradykinin (BK) response due to 18hours incubation with 1 μg/ml lipopolysaccharide (LPS; N = 8 to 12/group), without any amelioration of this effect by coincubation with 1.0mmL-NAME (left bars). Likewise, coincubation with 10 μm indomethacin failed to attenuate the effect of LPS (right bars). Kidney International 1998 54, 1063-1069DOI: (10.1046/j.1523-1755.1998.00090.x) Copyright © 1998 International Society of Nephrology Terms and Conditions