Article Figures & Data
Figures
- FIG 1
Timeline of experiments. T0, time zero.
- FIG 2
(A and D) Serial mean (±SEM) urine cAMP concentrations (A) and urine osmolality levels (D) in kidneys challenged with vasopressin or the diluent control alone and perfused under constant flow. (B, C, and E) Serial mean changes from the baseline (±SEM) for fractional free water reabsorption (B), the urine flow rate (C), and fractional sodium reabsorption (E). The numbers of kidneys employed in these experiments and baseline data for parameters shown as serial changes are noted in Tables 1 and 2, respectively.
- FIG 3
(A and D) Serial mean (±SEM) urine cAMP concentrations (A) and urine osmolality levels (D) in kidneys challenged with ET or PA alone and perfused under constant flow. (B, C, and E) Serial mean changes from the baseline (±SEM) for fractional free water reabsorption (B), the urine flow rate (C), and fractional sodium reabsorption (E). The numbers of kidneys employed in these experiments and baseline data for parameters shown as serial changes are noted in Tables 1 and 2, respectively.
- FIG 4
(A and D) Serial mean (±SEM) urine cAMP concentrations (A) and urine osmolality levels (D) in kidneys challenged with ET or PA alone and perfused under constant pressure. (B, C, and E) Serial mean changes from the baseline (±SEM) for fractional free water reabsorption (B), the urine flow rate (C) and fractional sodium reabsorption (E). The numbers of kidneys employed in these experiments and baseline data for parameters shown as serial changes are noted in Tables 1 and 2, respectively.
- FIG 5
Serial GFRs for experiments with kidneys challenged with ET or PA alone and perfused under constant flow (A) or constant pressure (B). The numbers of kidneys employed in these experiments and baseline data for parameters shown as serial changes are noted in Tables 1 and 2, respectively. * and ** denote levels of significance for overall effects between study groups and for time interactions between groups, respectively.
- FIG 6
(A and D) Serial mean (±SEM) urine cAMP concentrations (A) and urine osmolality levels (D) in kidneys challenged with ET, treated with raxibacumab or a nonspecific control MAb, and perfused under constant flow. (B, C, and E) Serial mean changes from the baseline (±SEM) for fractional free water reabsorption (B), the urine flow rate (C), and fractional sodium reabsorption (E). The numbers of kidneys employed in these experiments and baseline data for parameters shown as serial changes are noted in Tables 1 and 2, respectively.
- FIG 7
(A and D) Serial mean (±SEM) urine cAMP concentrations (A) and urine osmolality levels (D) in kidneys challenged with ET, treated with adefovir or a diluent, and perfused under constant flow. (B, C, and E) Serial mean changes from the baseline (±SEM) for fractional free water reabsorption (B), the urine flow rate (C), and fractional sodium reabsorption (E). The numbers of kidneys employed in these experiments and baseline data for parameters shown as serial changes are noted in Tables 1 and 2, respectively.
- FIG 8
Sample photomicrographs (A, C, E, G, I, and K) and AQP2 fluorescence intensity normalized to the maximum intensity for each image plotted along a 10-μm line crossing tubular cells from extraluminal to intraluminal regions in five selected tubules for each of the six kidneys (B, D, F, H, J, and L). In the photomicrographs, AQP2 staining is shown in pink, while nuclear staining and autofluorescence are shown in gray. Shown are photomicrographs and fluorescence intensity data for one kidney each challenged with AVP (A and B) or HEPES (AVP diluent) (C and D) and for two kidneys each challenged with edema toxin (ET for kidney 1 [ET 1] [E and F] and ET 2 [G and H]) or protective antigen alone (PA for kidney 1 [PA 1] [I and J] and PA 2 [K and L]). While the fluorescence intensity was dispersed widely along the plotted lines across tubules from kidneys challenged with HEPES and PA, this intensity tended to be localized over a short distance for tubules from AVP- and ET-challenged kidneys. As presented in Results, consistent with this narrow localization with ET but not PA, free AQP2 levels were significantly higher in urine from ET-perfused kidneys than in urine from PA-perfused kidneys at 90 and 120 min of perfusion (5.52 ± 1.06 ng/ml [n = 14 kidneys] versus 1.51 ± 0.44 ng/ml [n = 12] [means ± SEM, averaged over the two time points]; P = 0.0001).
Tables
- TABLE 1
Summary of dosages of challenges and treatments and numbers of kidneys employed in the isolated perfused kidney experiments
Expt Method of perfusion Challenge Treatment No. of kidneys tested Type Dosea Drug Dose Effect of challenge with vasopressin (AVP) Constant flow AVP 0.025 μM 3 Diluent 0 5 Constant flow AVP 0.05 μM 6 Diluent 0 7 Constant flow AVP 0.1 μM 5 Diluent 0 μg/ml 5 Effect of ET challenge with constant flow Constant flow PA 2.0 μg/ml 7 ET 1.0 μg/ml 7 Constant flow PA 4.0 μg/ml 7 ET 2.0 μg/ml 6 Effect of ET challenge with constant pressure Constant pressure PA 1.0 μg/ml 9 ET 0.5 μg/ml 9 Constant pressure PA 2.0 μg/ml 6 ET 1.0 μg/ml 6 Effect of raxibacumab treatment with ET challenge Constant flow ET 1.0 μg/ml Raxibacumab 10×b 8 ET 1.0 μg/ml Control 0 6 Constant flow ET 1.0 μg/ml Raxibacumab 50× 3 ET 1.0 μg/ml Control 0 3 Effect of adefovir treatment with ET challenge Constant flow ET 1.0 μg/ml Adefovir 20 μM 7 ET 1.0 μg/ml Control 0 5 Constant flow ET 2.0 μg/ml Adefovir 20 μM 4 ET 2.0 μg/ml Control 0 4 - TABLE 2
Baseline measurementsa
Parameter Mean value ± SD Effect of challenge with vasopressin (AVP) Effect of ET challenge with constant pressure Effect of Raxi with ET challenge Effect of adefovir with ET challenge Effect of ET challenge with constant flow Diluent AVP PA ET ET + NS-Ab ET + Raxi ET + diluent ET + adefovir PA ET Flow (ml/min) 32.8 ± 1.0 31.4 ± 0.9 29.3 ± 1.3 27.5 ± 0.8 33.1 ± 1.7 36.1 ± 1.6 30.8 ± 0.7 30.8 ± 0.9 29.5 ± 0.9 30.4 ± 0.6 Perfusion pressure (mm Hg) 101.8 ± 1.4 100.7 ± 0.9 99.9 ± 0.9 100.8 ± 0.8 98.8 ± 0.5 97.7 ± 1.0 100.9 ± 1.0 101.8 ± 1.0 101.1 ± 1.6 100.9 ± 1.6 GFR (ml/min) 0.83 ± 0.03 0.88 ± 0.05 0.71 ± 0.05 0.77 ± 0.06 0.77 ± 0.04 0.83 ± 0.04 0.75 ± 0.03 0.79 ± 0.04 0.75 ± 0.03 0.77 ± 0.03 Fractional free water reabsorption 0.81 ± 0.01 0.81 ± 0.01 0.88 ± 0.01 0.88 ± 0.01 0.83 ± 0.01 0.84 ± 0.01 0.84 ± 0.01 0.84 ± 0.01 0.84 ± 0.01 0.83 ± 0.02 Urine output (μl/min) 157.2 ± 10.9 168.1 ± 11.1 86.6 ± 8.4 95.4 ± 10.5 132.8 ± 13.9 135.7 ± 11.0 123.5 ± 7.8 125.7 ± 9.6 126 ± 15 130 ± 15 Fractional sodium reabsorption 0.82 ± 0.01 0.81 ± 0.01 0.88 ± 0.01 0.88 ± 0.01 0.83 ± 0.01 0.84 ± 0.01 0.84 ± 0.01 0.85 ± 0.01 0.84 ± 0.02 0.84 ± 0.02 ↵a Raxi, raxibacumab; NS-Ab, nonspecific antibody.
