Volume 281, October 2001
Volume 25, October 2001

Pages L993-L1000, 2001: C. Song, A. B. Al-Mehdi, and A. B. Fisher. "An immediate endothelial cell signaling response to lung ischemia." On pages L994, L997, and L998 (Figs. 1, A-C, 2, A-C, and 3, A-C, respectively), the figures should have appeared in color. In these figures, panels D and E are correctly printed in black and white. The correct figures and their legends follow.

View larger version (100K): [in this window] [in a new window]   Fig. 1.   4-{2-[6-(Dioctylamino)-2-naphthalenyl]ethenyl}1-(3-sulfopropyl)-pyridinium (di-8-ANEPPS) fluorescence in subpleural endothelial cells in situ in the intact rat lung. Lungs were perfused with di-8-ANEPPS for 30 min of equilibration, and then images were acquired, usually at 1-s intervals, before and after ischemia (global cessation of flow). Images are in pseudocolor with the intensity scale shown in C. Nos. in the images indicate time in seconds for the control period (negative numbers) or ischemic period (positive numbers for the same lung, with start of ischemia at 0. A: endothelial response to ischemia. Lungs were perfused under control conditions and then subjected to abrupt ischemia. B: effect of a K+ channel activator. Lemakalim (30 µM) was added to the perfusate during the 30-min equilibration period. C: quantitation of time course membrane potential with ischemia. Fluorescence intensity of 3 lungs (each representing the averaged value for 4-7 endothelial cells) for control and lemakalim-treated lungs was plotted as means ± SE. In inset, images of di-8-ANEPPS-stained vascular endothelial cells were taken as a stream (18 frames/s) for up to 5 s to increase resolution during the 1st s. D: calibration of di-8-ANEPPS fluorescence change by perfusion of isolated lungs with elevated K+ to induce membrane depolarization. Cells were loaded with di-8-ANEPPS and perfused with Krebs-Ringer bicarbonate (KRB; 5.9 mM K+) to obtain a baseline. The perfusate was abruptly changed to modified KRB with 7.5, 10, or 12 mM K+. Each curve represents the mean response of 3 endothelial cells from 1 lung. E: mean ± SE plot from 3 different lungs for each K+ concentration to calibrate di-8-ANEPPS fluorescence change. Con, control.

View larger version (109K): [in this window] [in a new window]   Fig. 2.   Amplex Red fluorescence in subpleural microvasculature in the intact rat lung in situ. Images were acquired as a stream (18 frames/s) for 12 s before and 11 s after ischemia. Nos. in the images indicate time in seconds for the control period (negative numbers) or ischemic period (positive numbers) for the same lung, with start of ischemia at 0. A: effect of ischemia in lungs without additions. B: effect of catalase. Lungs were perfused with catalase (1,000 U/ml) during the equilibration period. C: effect of extracellular Ca2+. Lungs were perfused with Ca2+-free buffer with added 1 mM EGTA during the equilibration period. D: time course of intravascular H2O2 with ischemia. Images were selected at 0.5-s intervals for quantification. Data are means ± SE of fluorescence intensity for 3 lungs for each condition. E: longer duration of ischemia. Images were acquired at 30-s intervals for 30 min. Note the different time scale compared with D.

View larger version (151K): [in this window] [in a new window]   Fig. 3.   Effect of ischemia on fluo 3 fluorescence in lung endothelial cells in situ. Fluo 3 was preperfused for 30 min before the onset of ischemia. Nos. in the images indicate time in seconds for the control period (negative numbers) or ischemic period (positive numbers) for the same lung, with start of ischemia at 0. A: fluo 3 fluorescence with ischemia under control conditions. B: effect of Ca2+-free perfusion on changes in fluo 3 fluorescence with ischemia. Lungs were perfused with Ca2+-free medium containing 1 mM EGTA. C: effect of depletion of intracellular Ca2+ stores on changes in fluo 3 fluorescence with ischemia. Thapsigargin (TG; 1 µM) was administered in the perfusate for 30 min during the equilibration period. D: time course for changes in intracellular Ca2+ with ischemia. Each data point represents the mean ± SE for 3 lungs for each condition.