Models of Renal Insufficiency: The Anti‐Thy‐1.1 Model of Acute Proliferative Glomerulonephritis

Glenn A. Reinhart1, Bryan F. Cox1

1 Abbott Laboratories, Abbott Park, Illinois
Publication Name:  Current Protocols in Pharmacology
Unit Number:  Unit 5.21
DOI:  10.1002/0471141755.ph0521s07
Online Posting Date:  May, 2001
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Abstract

This unit describes the rat anti‐Thy‐1.1 model of acute proliferative glomerulonephritis for the study of chronic renal insufficiency. A procedure is detailed for the induction of glomerulonephritis in rats as well as measurement of daily urinary excretion of protein, which is a convenient, primary screening tool. The unit also provides methods for assessment of glomerular filtration rate and effective renal plasma flow in anesthetized rats with anti‐Thy‐1.1‐induced renal insufficiency.

     
 
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Table of Contents

  • Basic Protocol 1: Daily Urinary Excretion of Protein as an Index of Renal Disease in Rats with Acute Proliferative Glomerulonephritis
  • Basic Protocol 2: Measurement of Glomerular Filtration Rate and Effective Renal Plasma Flow in Anesthetized Rats with Proliferative Glomerulonephritis
  • Support Protocol 1: Calculation of Glomerular Filtration Rate and Renal Plasma Flow
  • Reagents and Solutions
  • Commentary
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1: Daily Urinary Excretion of Protein as an Index of Renal Disease in Rats with Acute Proliferative Glomerulonephritis

  Materials
  • Male Wistar rats (130 to 150 g)
  • 10 mg/ml sodium azide solution
  • Test compound solution
  • 1 mg/kg/0.2 ml anti‐Thy‐1.1 antibody solution (see recipe)
  • 0.9% (w/v) NaCl, sterile
  • 50 mg/ml sodium Brevital (Eli Lilly)
  • Metabolism cages (Nalgene)
  • Urine collection tubes (e.g., 75 ml), supplied with metabolism cages
  • Animal balance, accurate to 1.0 g
  • 1‐ml syringes
  • 30‐G needles
  • 45°C water bath
  • 37°C warming pad
  • Additional reagents and equipment for Bradford or Lowry protein assay ( appendix 3A)
CAUTION: Sodium azide is a toxic substance, and should be handled appropriately.NOTE: Due to possible variation in specific activity of the ascites‐antibody between production lots, it is advisable to dedicate specific production lots to specific studies.

Basic Protocol 2: Measurement of Glomerular Filtration Rate and Effective Renal Plasma Flow in Anesthetized Rats with Proliferative Glomerulonephritis

  Materials
  • Male Wistar rats (200 to 225 g)
  • Inactin (Research Biochemicals)
  • 0.9% (w/v) NaCl
  • Isotope solution (see recipe)
  • Sodium pentobarbital
  • Water‐compatible scintillation fluid (Fisher ScintiVerse or equivalent)
  • Venous and arterial catheters (PE‐50; 10 to 14 cm for arterial catheter)
  • Urinary bladder catheter (PE‐90; 4 to 4.5 cm, with flared tip)
  • 500‐ to 1000‐µl urine collection tubes
  • 18‐G needle
  • Cardiovascular workstation (Modular Instruments or equivalent) equipped with MA2 transducer amplifiers for measurement of mean arterial pressure and heart rate
  • Warming pad/table capable of maintaining rat's core temperature at 37° ± 1°C
  • Rodent thermistor rectal probe (YSI Precision 4000A or equivalent)
  • Small animal surgical tools and equipment
  • Pressure transducer (Transpac II, Abbott Laboratories, or equivalent)
  • 23‐G needle hubs
  • PE tubing (PE‐50, ‐90, and ‐205)
  • Syringe infusion pump
  • Microhematocrit tubes and hematocrit centrifuge
  • Hematocrit tube sealing clay
  • Diamond pencil
  • 5‐ml scintillation vials with caps
  • Scintillation counter capable of simultaneous measurement of 14C and 3H
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Figures

Videos

Literature Cited

Literature Cited
   Bagchus, W.M., Hoedemaeker, P.J., Rozing, J., and Bakker, W.W. 1986. Glomerulonephritis induced by monoclonal anti‐Thy‐1.1 antibodies. A sequential histological and ultrastructural study in the rat. Lab. Invest. 55:680‐687.
   Border, W.A., Okuda, S., Languino, L.R., Sporn, M.B., and Ruoslahti, E. 1990. Suppression of experimental glomerunephritis transforming growth factor β1. Nature 346:371‐374.
   Border, W.A., Noble, N.A., Yamamoto, T., Harper, J.R., Yamaguchi, Y., Pierschbacher, M.D., and Ruoslahti, E. 1992. Natural inhibitor of transforming growth factor‐β protects against kidney disease. Nature 360:361‐364.
   Gretz, N., Waldherr, R., and Strauch, M. 1993. The remnant kidney model In Experimental and Gentic Rat Models of Chronic Renal Failure (N. Gretz and M. Strauch, eds.) pp. 1‐28. Karger, Basel.
   Morris, R.J. and Ritter, M.A. 1980. Association of Thy‐1 cell surface differentiation antigen with certain connective tissues in vivo. Cell Tissue Res. 206:459‐475.
   Okuda, S., Nakamura, T., Yamamoto, T., Ruoslahti, E., and Border, W.A. 1991. Dietary protein restriction rapidly reduces transforming growth factor β1 expression in experimental glomerulonephritis. Proc. Natl. Acad. Sci. U.S.A. 88:9765‐9769.
   Reif, A.E. and Allen, J.M.V. 1964. The AKR thymic antigen and its distribution in leukaemias and nervous tissues. J. Exp. Med. 120:413‐433.
   Reinhart, G.A., Kramer, D., Opgenorth, T., Wegner, C.D., and Cox, B.F. 1998. Renal functional effects of ETA receptor blockade in the Thy‐1 model of proliferative glomerulonephritis. J. Am. Soc. Nephrol. 9A‐3175.
   Stahl, R.A.K., Thaiss, F., Oberle, G., Brecht, H.M., Schoeppe, W., Wenzel, U., and Helmchen, U.M. 1991. The platelet activating factor receptor antagonist WEB 2170 improves glomerular hemodynamics and morphology in a proliferative model of mesangial cell injury. J. Am. Soc. Nephrol. 2:37‐44.
   Steinmann‐Niggli, K., Ziswiler, R., Kung, M., and Marti, H.P. 1998. Inhibition of matrix metalloproteinases attenuates anti‐Thy‐1.1 nephritis. J. Am. Soc. Nephrol. 9:397‐407.
   Yamamoto, T. and Wilson, C.B. 1987. Quantitative and qualitative studies of antibody‐induced mesangial cell damage in the rat. Kidney Int. 32:514‐525.
   Yamamoto, T., Mundy, C.A., Wilson, C.B., and Blantz, R.C. 1991. Effect of mesangial cell lysis and proliferation on glomerular hemodynamics in the rat. Kidney Int. 40:705‐713.
   Ziswiler, R., Steinmann‐Nigle, L., Kappeler, A., Daniel, C., and Marti, H.P. 1998. Mycophenolic acid: A new approach to the therapy of experimental mesangial proliferative glomerulonephritis. J. Am. Soc. Nephrol. 9:2055‐2066.
Key References
   Vander, A.J. 1995. Renal Physiology, 5th ed. McGraw‐Hill, New York.
  A basic handbook that reviews the fundamental principles of renal physiology, including principles of clearance measurements.
   Levinsky, N.G. and Lieberthal, W. 1992. Clearance Techniques. In Handbook of Physiology, Section 8: Renal Physiology Vol. I (E.E. Windhager, ed.) pp. 227‐248. Oxford University Press, New York, Oxford.
  Reviews clearance techniques, substances used, and related techniques for measurement of renal hemodynamics.
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