Streptozotocin‐Induced Diabetic Models in Mice and Rats

Kenneth K. Wu1, Youming Huan2

1 Merck Research Laboratories, Rahway, New Jersey, 2 Mount Sinai School of Medicine, New York, New York
Publication Name:  Current Protocols in Pharmacology
Unit Number:  Unit 5.47
DOI:  10.1002/0471141755.ph0547s40
Online Posting Date:  March, 2008
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Abstract

Streptozotocin (STZ) is an antibiotic that can cause pancreatic β‐cell destruction, so it is widely used experimentally as an agent capable of inducing insulin‐dependent diabetes mellitus (IDDM), also known as type 1 diabetes mellitus (T1DM). This unit describes protocols for the production of insulin deficiency and hyperglycemia in mice and rats, using STZ. These models for diabetes can be employed for assessing the mechanisms of T1DM, screening potential therapies for the treatment of this condition, and evaluation of therapeutic options. Curr. Protoc. Pharmacol. 40:5.47.1‐5.47.14. © 2008 by John Wiley & Sons, Inc.

Keywords: streptozotocin; type 1 diabetes mellitus; hyperglycemia; insulin deficiency; insulitis; mouse; rat

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Induction of Type 1 Diabetes Mellitus in Mice using Repeated Low Doses of Streptozotocin
  • Alternate Protocol 1: Induction of Type 1 Diabetes Mellitus in Mice using a Single, High Dose of Streptozotocin
  • Basic Protocol 2: Streptozotocin‐Induced Type 1 Diabetes Mellitus in Rats
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Induction of Type 1 Diabetes Mellitus in Mice using Repeated Low Doses of Streptozotocin

  Materials
  • C57BL/6 or CD‐1 male mice: 25 g, 8 to 12 weeks old (The Jackson Laboratory or Taconic); 12 to 20 per treatment group, recommended
  • Standard rodent chow diet (Harlan)
  • 50 mM sodium citrate (enzyme grade; Fisher) buffer, pH 4.5: prepared just before use
  • Streptozotocin (STZ; Sigma)
  • 10% (w/v) sucrose (Sigma): prepared just before use
  • Test compound(s)
  • Rodent cages
  • Temperature‐, humidity‐, and light‐controlled housing
  • 1.5‐ml microcentrifuge tubes
  • Aluminum foil
  • 1‐ml syringes
  • 25‐G needles
  • One Touch Basic blood glucose monitoring system (Lifescan)

Alternate Protocol 1: Induction of Type 1 Diabetes Mellitus in Mice using a Single, High Dose of Streptozotocin

  Materials
  • See protocol 1

Basic Protocol 2: Streptozotocin‐Induced Type 1 Diabetes Mellitus in Rats

  Materials
  • Sprague‐Dawley or Wistar male rats: ∼150 to 200 g, 8 to 10 weeks old (Charles River Breeding Laboratories); 10 to 16 per treatment group, recommended
  • Standard rodent chow diet (Harlan)
  • 50 mM sodium citrate (enzyme grade; Fisher) buffer, pH 4.5: prepared just before use
  • Streptozotocin (STZ; Sigma)
  • 10% (w/v) sucrose (Sigma): prepared just before use
  • 50 mM sodium citrate (enzyme grade; Fisher) buffer, pH 4.5: prepared just before use
  • Test compound(s)
  • Rodent cages
  • Temperature‐, humidity‐, and light‐controlled housing
  • 1.5‐ml microcentrifuge tubes
  • Aluminum foil
  • 3‐ml syringes
  • 23‐G needles
  • One Touch Basic blood glucose monitoring system (Lifescan)
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Figures

  •   FigureFigure 5.47.1 Schematic representation of the time course of multiple, low‐dose STZ‐induced diabetes in mice. Mice were treated with STZ (40 mg/kg) or without STZ (vehicle control) for 5 consecutive days. Typical changes in insulin secretion and blood glucose levels are illustrated.
  •   FigureFigure 5.47.2 A single, high dose of STZ causes diabetes in mice ( n = 20). Mice were treated with 200 mg/kg STZ or sodium citrate buffer vehicle (control). The effect of STZ on (A) nonfasting blood glucose level, (B) body weight, and (C) daily water intake at 3 weeks after the STZ injection. Data represent the mean ± SEM. * p < 0.001 versus control.
  •   FigureFigure 5.47.3 STZ‐induced hyperglycemia in rats. A single 65 mg/kg dose of STZ causes hyperglycemia in rats. Fasting blood glucose levels were monitored before and after the STZ injection on the indicated days.
  •   FigureFigure 5.47.4 Effect of daily treatment with inosine on STZ‐induced diabetes in mice. Daily treatment with 100 or 200 mg/kg inosine for 21 days decreased hyperglycemia (A) and incidence of diabetes (B) following multiple, low‐dose STZ (MLDS) treatment of the mice. Mice were either untreated (open circles); given daily doses of 200 mg/kg inosine alone (open squares); or treated with STZ (on days 1 to 5) in combination with vehicle (filled circles), 100 mg/kg inosine (filled triangles), or 200 mg/kg inosine (filled squares) starting on day 1. Diabetes incidence is expressed as a cumulative percentage of mice with a blood glucose ≥11 mmol/liter. Results are means ± SE for n = 20 mice in two separate experiments with 10 mice per experimental group. ** p < 0.01 compared with vehicle‐treated mice; † p < 0.05; †† p < 0.01 compared with MLDS‐treated mice. Reproduced from Mabley et al., with permission from The Feinstein Institute for Medical Research.
  •   FigureFigure 5.47.5 Restoration of ß‐cell function in STZ‐induced diabetic mice. (A) Blood glucose (BG) levels of single high‐dose STZ‐induced diabetic (open circles) or restored (filled squares) mice. (B) Mean BG levels (±SEM) were calculated from each group. * and ** indicate statistical differences (ANOVA) between the restored ( n = 14) and the normal ( n = 10) or diabetic ( n = 34) groups, respectively. Reproduced from Yin et al., with permission from the American Diabetes Association.
  •   FigureFigure 5.47.6 Effect of valsartan on plasma glucose concentration in STZ‐induced diabetic rats. The diabetic rats were treated with valsartan 2 weeks after STZ‐injection. Values of mean ± SE were obtained from each group of eight animals. *p < 0.05 and **p < 0.01 versus data from animals treated with vehicle (0 mg/kg valsartan). Reproduced from Chan et al., with permission from Lippincott, Williams, & Wilkins.

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Literature Cited

   Arison, R.N., Ciaccio, E.I., Glitzer, M.S., Cassaro, J.A., and Pruss, M.P. 1967. Light and electron microscopy of lesions in rats rendered diabetic with streptozotocin. Diabetes 16: 51‐56.
   Bond, J.S., Failla, M.L., and Unger, D.F. 1983. Elevated manganese concentration and arginase activity in livers of streptozotocin‐induced diabetic rats. J. Biol. Chem. 258: 8004‐8009.
   Bonnevie‐Nielsen, V., Steffes, M.W., and Lernmark, A. 1981. A major loss in islet mass and B‐cell function precedes hyperglycemia in mice given multiple low doses of streptozotocin. Diabetes 30: 424‐429.
   Breyer, M.D., Bottinger, E., Brosius, F.C. 3rd, Coffman, T.M., Harris, R.C., Heilig, C.W., and Sharma, K., AMDCC. 2005. Mouse models of diabetic nephropathy. J. Am. Soc. Nephrol. 16: 27‐45.
   Brondum, E., Nilsson, H., and Aalkjaer, C. 2005. Functional abnormalities in isolated arteries from Goto‐Kakizaki and streptozotocin‐treated diabetic rat models. Horm. Metab. Res. 37: 56‐60.
   Chan, P., Wong, K.L., Liu, I.M., Tzeng, T.F., Yang, T.L., and Cheng, J.T. 2003. Antihyperglycemic action of angiotensin II receptor antagonist, valsartan, in streptozotocin‐induced diabetic rats. J. Hypertens. 21: 761‐769.
   Flood, J.F., Mooradian, A.D., and Morley, J.E. 1990. Characteristics of learning and memory in streptozotocin‐induced diabetic mice. Diabetes 39: 1391‐1398.
   Ganda, O.P., Rossini, A.A., and Like, A.A. 1976. Studies on streptozotocin diabetes. Diabetes 25: 595‐603.
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   Huang, F. and Wu, W. 2005. Antidiabetic effect of a new peptide from Squalus mitsukurii liver (S‐8300) in streptozocin‐induced diabetic mice. J. Pharm. Pharmacol. 57: 1575‐1580.
   Junod, A., Lambert, A.E., Stauffacher, W., and Renold, A.E. 1969. Diabetogenic action of streptozotocin: Relationship of dose to metabolic response. J. Clin. Invest. 48: 2129‐2139.
   Kolb, H. 1987. Mouse models of insulin dependent diabetes: Low‐dose streptozocin‐induced diabetes and nonobese diabetic (NOD) mice. Diabetes Metab. Rev. 3: 751‐778.
   Kolb‐Bachofen, V., Epstein, S., Kiesel, U., and Kolb, H. 1988. Low‐dose streptozocin‐induced diabetes in mice. Electron microscopy reveals single‐cell insulitis before diabetes onset. Diabetes 37: 21‐27.
   Kunjathoor, V.V., Wilson, D.L., and LeBoeuf, R.C. 1996. Increased atherosclerosis in streptozotocin‐induced diabetic mice. J. Clin. Invest. 97: 1767‐1773.
   Like, A.A. and Rossini, A.A. 1976. Streptozotocin‐induced pancreatic insulitis: New model of diabetes mellitus. Science 193: 415‐417.
   Mabley, J.G., Rabinovitch, A., Suarez‐Pinzon, W., Haskó, G., Pacher, P., Power, R., Southan, G., Salzman, A., and Szabó, C. 2003. Inosine protects against the development of diabetes in multiple‐low‐dose streptozotocin and nonobese diabetic mouse models of type 1 diabetes. Mol. Med. 9: 96‐104.
   Rossini, A.A., Appel, M.C., Williams, R.M., and Like, A.A. 1977. Genetic influence of the streptozotocin‐induced insulitis and hyperglycemia. Diabetes 26: 916‐920.
   Usuki, S., Ito, Y., Morikawa, K., Kise, M., Ariga, T., Rivner, M., and Yu, R.K. 2007. Effect of pregerminated brown rice intake on diabetic neuropathy in streptozotocin‐induced diabetic rats. Nutr. Metab. (Lond). 4: 25.
   Weide, L.G. and Lacy, P.E. 1991. Low‐dose streptozocin‐induced autoimmune diabetes in islet transplantation model. Diabetes 40: 1157‐1162.
   Wu, K.K. and Huan, Y. 2007. Diabetic atherosclerosis mouse models. Atherosclerosis. 191: 241‐249.
   Yin, D., Tao, J., Lee, D.D., Shen, J., Hara, M., Lopez, J., Kuznetsov, A., Philipson, L.H., and Chong, A.S. 2006. Recovery of islet beta‐cell function in streptozotocin‐induced diabetic mice: An indirect role for the spleen. Diabetes 55: 3256‐3263.
Internet Resource
   http://www.AMDCC.org
  Web site for Animal Models of Diabetic Complications Consortium (AMDCC), providing new animal models of diabetic complications, with the goal of identifying the most appropriate animal models to study the etiology, prevention, and treatment of diabetic complications.
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