Volume 55, Issue 3, Pages 793-807 (March 1999) Recovery following relief of unilateral ureteral obstruction in the neonatal rat  Robert L. Chevalier, Andrew Kim, Barbara A. Thornhill, Jennifer T. Wolstenholme  Kidney International  Volume 55, Issue 3, Pages 793-807 (March 1999) DOI: 10.1046/j.1523-1755.1999.055003793.x Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 1 Scheme of the experimental design for the five groups of rats studied. In group 1, rats were sham operated within the first 24 hours of life, and kidneys were removed for study five days later. In group 2, rats were subjected to unilateral ureteral obstruction (UUO) within the first 24 hours of life, and kidneys were removed for study five days later. In group 3, rats were subjected to sham operation during the first 24 hours of life, and kidneys were removed for study either 14 or 28 days later (19 or 33 days following the initial operation). In group 4, rats were subjected to UUO within the first 24 hours of life. The obstruction was released five days later, and kidneys were removed for study either 14 or 28 days later. In group 5, rats were subjected to UUO within the first 24 hours of life, and kidneys were removed for study either 14 or 28 days later. Kidney International 1999 55, 793-807DOI: (10.1046/j.1523-1755.1999.055003793.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 2 Immunochemical distribution of cytoplasmic proteins or markers of cellular proliferation or apoptosis in rat kidneys. (A–E) Distribution of immunoreactive renin in kidneys of rats from groups 1 through 5. Renin-containing vascular cells are brown. In sham-operated groups (A, group 1, and C, group 3, 33 days), renin is localized to the juxtaglomerular region of the afferent arteriole. In UUO groups (B, group 2, and E, group 5, 33 days), renin extends along the length of the afferent arteriole. In UUO followed by relief at five days (D, group 4, 33 days), renin distribution is intermediate between that of groups 3 and 5. (F) The localization of proliferating cells by proliferating cell nuclear antigen (PCNA) in obstructed kidney from rat in group 2 (19 days). Tubular cell nuclei stain brown (arrowheads). (G) The localization of apoptotic cells by TdT uridine-nick-end-label (TUNEL) technique from rat in group 5 (19 days): fragmented DNA in apoptotic nuclei stain brown (arrowheads). (H) The distribution of epidermal growth factor (EGF) in tubules from rat in group 3 (33 days; *). (I) The distribution of transforming growth factor-β1 (TGF-β1) in tubules from rat in group 5 (33 days; *). (J) The distribution of clusterin in tubules from rat in group 5 (33 days; *). (K) The distribution of vimentin in tubules from rat in group 5 (33 days; *). Kidney International 1999 55, 793-807DOI: (10.1046/j.1523-1755.1999.055003793.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 3 Length of afferent arteriole adjacent to glomerulus containing immunoreactive renin for rats in groups 1 and 2 (A) and in groups 3 through 5 (B). Data are mean ± SEM. Symbols are: (□) left kidney, five days (groups 1 and 2); (▪) left kidney, 33 days; (▩) right (contralateral) kidney. *P < 0.05 vs. sham; #P < 0.05 vs. contralateral kidney; +P < 0.05 vs. 5 days of relief. Kidney International 1999 55, 793-807DOI: (10.1046/j.1523-1755.1999.055003793.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 4 (A) Weighted glomerular maturation score in sham-operated and UUO rats five days after operation. (B) Glomerular tuft area 28 days after operation. (C) Relative number of glomeruli five days after operation. (D) The relative number of glomeruli 28 days after operation. Symbols are: (□) left kidney, five days (groups 1 and 2); (▨) right (contralateral) kidney; (▪) left kidney, 33 days; (▩) right (contralateral) kidney. Kidney International 1999 55, 793-807DOI: (10.1046/j.1523-1755.1999.055003793.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 5 Density of proliferating cells in kidneys, identified by proliferating cell nuclear antigen (PCNA). (A) PCNA-staining tubular cells five days after operation. (B) PCNA-staining tubular cells 14 days after operation. (C) PCNA-staining interstitial cells five days after operation. (D) PCNA-staining interstitial cells 14 days after operation. Symbols are: (□) left kidney, five days (groups 1 and 2); (▨) right (contralateral) kidney; (▪) left kidney, 33 days; (▩) right (contralateral) kidney. Kidney International 1999 55, 793-807DOI: (10.1046/j.1523-1755.1999.055003793.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 6 Density of apoptotic cells in kidneys, identified by TdT uridine-nick-end-label (TUNEL) technique. (A) Apoptotic tubular cells five days after operation. (B) Apoptotic tubular cells 14 days after operation. (C) Apoptotic interstitial cells five days after operation. (D) Apoptotic interstitial cells 14 days after operation. Symbols are: (□) left kidney, five days (groups 1 and 2); (▨) right (contralateral) kidney; (▪) left kidney, 33 days; (▩) right (contralateral) kidney. Kidney International 1999 55, 793-807DOI: (10.1046/j.1523-1755.1999.055003793.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 7 (A) Density of epidermal growth factor (EGF)-staining tubules five days after operation. (B) Density of EGF-staining tubules 28 days after operation (note scale is 100-fold reduced compared with A). (C) Density of atrophic tubules five days after operation. (D) Density of atrophic tubules 28 days after operation. Symbols are: (□) left kidney, five days (groups 1 and 2); (▨) right (contralateral) kidney; (▪) left kidney, 33 days; (▩) right (contralateral) kidney. Kidney International 1999 55, 793-807DOI: (10.1046/j.1523-1755.1999.055003793.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 8 (A) Relative tubular staining for transforming growth factor-β1 (TGF-β1) five days after operation. (B) Relative tubular staining for transforming growth factor-β1 (TGF-β1) 28 days after operation. (C) Density of clusterin-staining tubules five days after operation. (D) Density of clusterin-staining tubules 28 days after operation. (E) Density of vimentin-staining tubules 28 days after operation. Symbols are: (□) left kidney, five days (groups 1 and 2); (▨) right (contralateral) kidney; (▪) left kidney, 33 days; (▩) right (contralateral) kidney. Kidney International 1999 55, 793-807DOI: (10.1046/j.1523-1755.1999.055003793.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 9 Interstitial immunostaining for collagen (A–C), α-smooth muscle actin (D–F), and vimentin (G–I) for groups studied 28 days after operation. For each stain, sham-operated kidneys are on the left (A, D, and G); five-day relief kidneys are in the center (B, E, and H); and UUO kidneys are on the right (C, F, and I). Kidney International 1999 55, 793-807DOI: (10.1046/j.1523-1755.1999.055003793.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 10 (A) The relative area of interstitial collagen immunostaining 28 days after operation. (B) The relative area of interstitial α-smooth muscle actin immunostaining five days after operation. (C) The relative area of interstitial α-smooth muscle actin immunostaining 28 days after operation. (D) The relative area of interstitial vimentin immunostaining five days after operation. (E) The relative area of interstitial vimentin 28 days after operation. Symbols are: (□) left kidney, five days (groups 1 and 2); (▨) right (contralateral) kidney; (▪) left kidney, 33 days; (▩) right (contralateral) kidney. Kidney International 1999 55, 793-807DOI: (10.1046/j.1523-1755.1999.055003793.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Kidney International 1999 55, 793-807DOI: (10. 1046/j. 1523-1755. 1999 Kidney International 1999 55, 793-807DOI: (10.1046/j.1523-1755.1999.055003793.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Kidney International 1999 55, 793-807DOI: (10. 1046/j. 1523-1755. 1999 Kidney International 1999 55, 793-807DOI: (10.1046/j.1523-1755.1999.055003793.x) Copyright © 1999 International Society of Nephrology Terms and Conditions