Analyzing corticosterone metabolites in fecal samples of mice: a noninvasive technique to monitor stress hormones

Horm Behav. 2004 Jan;45(1):10-22. doi: 10.1016/j.yhbeh.2003.07.002.


In small animals like mice, the monitoring of endocrine functions over time is constrained seriously by the adverse effects of blood sampling. Therefore, noninvasive techniques to monitor, for example, stress hormones in these animals are highly demanded in laboratory as well as in field research. The aim of our study was to evaluate the biological relevance of a recently developed technique to monitor stress hormone metabolites in fecal samples of laboratory mice. In total, six experiments were performed using six male and six female mice each. Two adrenocorticotropic hormone (ACTH) challenge tests, two dexamethasone (Dex) suppression tests and two control experiments [investigating effects of the injection procedure itself and the diurnal variation (DV) of glucocorticoids (GCs), respectively] were conducted. The experiments clearly demonstrated that pharmacological stimulation and suppression of adrenocortical activity was reflected accurately by means of corticosterone metabolite (CM) measurements in the feces of males and females. Furthermore, the technique proved sensitive enough to detect dosage-dependent effects of the ACTH/Dex treatment and facilitated to reveal profound effects of the injection procedure itself. Even the naturally occurring DV of GCs could be monitored reliably. Thus, our results confirm that measurement of fecal CM with the recently established 5alpha-pregnane-3beta,11beta,21-triol-20-one enzyme immunoassay is a very powerful tool to monitor adrenocortical activity in laboratory mice. Since mice represent the vast majority of all rodents used for research worldwide and the number of transgenic and knockout mice utilized as animal models is still increasing, this noninvasive technique can open new perspectives in biomedical and behavioral science.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Adrenal Cortex / drug effects
  • Adrenal Cortex Function Tests
  • Adrenocorticotropic Hormone / pharmacology
  • Analysis of Variance
  • Animals
  • Circadian Rhythm
  • Corticosterone / analysis*
  • Corticosterone / metabolism
  • Dexamethasone / pharmacology
  • Feces / chemistry*
  • Female
  • Immunoenzyme Techniques / methods*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Models, Animal
  • Reproducibility of Results
  • Stress, Psychological / metabolism*


  • Dexamethasone
  • Adrenocorticotropic Hormone
  • Corticosterone