Volume 69, Issue 11, Pages (June 2006)

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Volume 69, Issue 11, Pages 2013-2021 (June 2006) Subtotal nephrectomy impairs ischemia-induced angiogenesis and hindlimb re-perfusion in rats J. Jacobi, M. Porst, N. Cordasic, B. Namer, R.E. Schmieder, K.-U. Eckardt, K.F. Hilgers Kidney International Volume 69, Issue 11, Pages 2013-2021 (June 2006) DOI: 10.1038/sj.ki.5000448 Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 1 Histomorphology of skeletal muscle and blood vessels. (a) Skeletal muscle histomorphology (M. soleus) 2 weeks after induction of limb ischemia. Arrows indicate atrophied myocytes with centralized nuclei (original magnification, × 20, hematoxylin and eosin (H&E) staining). (b) Analysis of collateral vessels by light microscopy: collateral vessel within ischemic skeletal muscle (M. gracilis) of a sham-treated rat. Blood vessels are surrounded by an inflammatory infiltrate and myocytes with centralized nuclei (original magnification, × 20, H&E staining). (c) Analysis of collateral vessels by fluorescence microscopy: collateral vessel within ischemic skeletal muscle (M. gracilis) of a sham-treated rat; (red) CD31 and (green) α-smooth muscle actin (original magnification, × 40). Kidney International 2006 69, 2013-2021DOI: (10.1038/sj.ki.5000448) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 2 Neovascularization of skeletal muscle. (a) Percent increase of capillary density (capillaries/myocyte) in ischemic hindlimbs vs non-ischemic hindlimbs 14 days after induction of ischemia. Angiogenesis is significantly impaired in rats that underwent renal mass reduction. (b) Percent increase of capillary area (capillary area/100 myocytes) in ischemic hindlimbs vs non-ischemic hindlimbs 14 days after induction of ischemia. *P<0.05; **P<0.01; ***P<0.001. Kidney International 2006 69, 2013-2021DOI: (10.1038/sj.ki.5000448) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 3 Representative images of capillary density in non-ischemic and ischemic calf muscle sections. Top panel: CD31 immunofluorescence (capillaries in red); bottom panel: double fluorescent staining for (red) CD31 and (blue) laminin. (a) Capillary density of non-ischemic muscle of a rat with renal mass reduction. (b) Capillary density of ischemic muscle of a rat with renal mass reduction. (c) Capillary density of ischemic muscle of a sham-treated rat. Hindlimb ischemia causes neovascularization (angiogenesis) in both treatment groups with a greater response in rats without renal impairment. Kidney International 2006 69, 2013-2021DOI: (10.1038/sj.ki.5000448) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 4 (a) Laser Doppler perfusion imaging. Flux images of a sham (top panel) and nephrectomized (bottom panel) rat before surgery and on days 1 and 14 post-surgery. Hindpaw re-perfusion is impaired in red rectangles subtotally nephrectomized rats (red rectangle). (b) LDPI ratio (ischemic/non-ischemic hindpaw). After induction of hindlimb ischemia, there was a dramatic reduction in hindpaw perfusion in both sham as well as nephrectomized rats. At 2 weeks after surgery, perfusion ratio was significantly lower in rats with renal mass reduction as opposed to sham-treated animals. *P<0.05. Kidney International 2006 69, 2013-2021DOI: (10.1038/sj.ki.5000448) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 5 Expression of VEGF and VEGFR-1 and -2 mRNA in non-ischemic and ischemic muscles (reverse transcription-polymerase chain reaction from M. tibialis anterior). Top panel: Relative copy numbers; bottom panel: relative change of expression in ischemic vsnon-ischemic limbs. Kidney International 2006 69, 2013-2021DOI: (10.1038/sj.ki.5000448) Copyright © 2006 International Society of Nephrology Terms and Conditions