Glucose Tolerance Tests for Systematic Screening of Glucose Homeostasis in Mice

Jan Rozman1, Birgit Rathkolb1, Susanne Neschen1, Helmut Fuchs2, Valérie Gailus‐Durner2, Martin Klingenspor3, Eckhard Wolf4, Martin Hrabě de Angelis5

1 These authors contributed equally to this work, 2 Institute of Experimental Genetics, German Mouse Clinic, Helmholtz‐Zentrum München, German Center for Environmental Health, Neuherberg, 3 Molecular Nutritional Medicine, Else‐Kröner Fresenius Center for Food Sciences & ZIEL–Research Center for Nutrition and Food Sciences, Technische Universität München, Freising, 4 Institute of Molecular Animal Breeding and Biotechnology, Gene Center, Ludwig‐Maximilians‐Universität München, Munich, 5 Corresponding author: Martin Hrabě de Angelis (hrabe@helmholtz-muenchen.de)
Publication Name:  Current Protocols in Mouse Biology
Unit Number:   
DOI:  10.1002/9780470942390.mo140111
Online Posting Date:  March, 2015
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Abstract

This article presents a detailed description of intraperitoneal and oral glucose tolerance tests in mice. The former is widely used in initial high‐throughput phenotyping of mutant mice to assess a diabetic phenotype and alterations in glucose homeostasis. Each protocol provides a comprehensive description of each step in the workflow, including variation of the standard protocol under particular circumstances (e.g., sensitivity to food deprivation, excessive deviations in body composition, or need for extra blood samples for additional analyses). We also describe how reduction of body mass and body temperature can be used as additional readouts to monitor metabolic function in response to food deprivation. © 2015 by John Wiley & Sons, Inc.

Keywords: glucose tolerance test; IPGTT; OGTT; type 2 diabetes mellitus; mouse models; glucose intolerance; thermovision; high‐throughput phenotyping

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Intraperitoneal Glucose Tolerance Test (IPGTT)
  • Basic Protocol 2: Oral Glucose Tolerance Test (OGTT)
  • Support Protocol 1: Evaluate Mouse Capacity to Compensate for Energy Shortage Induced by Food Deprivation
  • Commentary
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1: Intraperitoneal Glucose Tolerance Test (IPGTT)

  Materials
  • Mice (matched for sex, genotype, and body mass)
  • 20% aqueous glucose solution, sterile (B. Braun Melsungen AG)
  • Sterile 1‐ml syringes, Omnican‐F, 0.01 ml scale (B. Braun Melsungen AG)
  • Needles, Sterican cannula, 27G × 12 mm or 26G × 20 mm (B. Braun Melsungen AG), length depends on mouse size and body fat content (Fig.  A)
  • Protocol sheets [one per mouse; for example, see Fig.  : the protocol sheet can be a printed paper form or an electronic spreadsheet that can be uploaded to a laboratory information management system (LIMS)]
  • Scales, e.g., Kern 440‐47 N (Kern & Sohn GmbH) that can be linked to a computer for automated data storage (e.g., Kern SHM‐01 software, Kern & Sohn GmbH)
  • Mouse cages with clean bedding only (no food in hopper or bottom of the cage) to be used during food deprivation
  • Mouse cages without bedding (one cage per animal) to be used during the test
  • Blood glucose meters (e.g., ACCU‐CHECK® Aviva, Roche Diagnostics, or Contour®, Bayer) and corresponding test strips (Fig.  B)
  • Timers (ideally one per mouse)
  • Cage grid
  • Sterile scalpels or razor blades

Basic Protocol 2: Oral Glucose Tolerance Test (OGTT)

  Materials
  • Mice (matched for sex, genotype, and body mass)
  • 20% aqueous glucose solution, sterile (B. Braun Melsungen AG)
  • Sterile 1‐ml syringes, Omnican‐F, 0.01 ml scale (B. Braun Melsungen AG)
  • Curved or straight gavage needles with ball‐tip for oral glucose application (Fig.  A), size of needle depends on mouse size (e.g., VWR, cat. no. 20068‐636 for adult; VWR, cat. no. 20068‐608 for newborn)
  • Protocol sheets (one per mouse)
  • Scales, e.g., Kern 440‐47 N (Kern & Sohn GmbH), that can be linked to a computer for automated data storage (e.g., Kern SHM‐01 software, Kern & Sohn GmbH)
  • Mouse cages with clean bedding only (no food in hopper or bottom of the cage) to be used during food deprivation
  • Mouse cages without bedding (one cage per animal) to be used during the test
  • Blood glucose meters (e.g., ACCU‐CHECK® Aviva, Roche Diagnostics, or Contour®, Bayer) and corresponding test strips (Fig.  B)
  • Timers (ideally one per mouse)
  • Cage grid
  • Sterile scalpel or razor blades

Support Protocol 1: Evaluate Mouse Capacity to Compensate for Energy Shortage Induced by Food Deprivation

  Materials
  • Mice
  • Thermovision camera, e.g., FLIR I60 (TOPA GmbH; Fig.  A, left panel)
  • Animal scales, e.g., Kern 440‐47 N (Kern & Sohn GmbH)
  • Disinfectant for cleaning and disinfection of surfaces that were in contact with mice
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Figures

Videos

Literature Cited

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Key References
  Joslin, E.P. and Kahn, C.R. 2005. Joslin's diabetes mellitus, 14th ed. Lippincott Williams & Wilkins. Philadelphia, PA.
  This book provides detailed background information on diabetes mellitus and glucose homeostasis.
Internet Resources
  http://www.endotext.org/
  This site provides comprehensive, authoritative, and constantly updated information on endocrine diseases, covering diabetes mellitus and other carbohydrate homeostasis diseases.
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