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Hair cortisol: a parameter of chronic stress? Insights from a radiometabolism study in guinea pigs

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Abstract

Measurement of hair cortisol has become popular in the evaluation of chronic stress in various species. However, a sound validation is still missing. Therefore, deposition of radioactivity in hair and excretion into feces and urine after repeated injection of 3H-cortisol was studied in guinea pigs (n = 8). Each animal was given intraperitoneally 243.6 kBq 3H-cortisol/day on 3 successive days. After the first injection, all voided excreta were collected for 3 days. After the second injection, hair was shaved off the animals’ back and newly grown hair was obtained on day 7. Following methanol extraction, radiolabeled and unlabeled glucocorticoid metabolites (GCM) in fecal and hair samples were characterized by high-performance liquid chromatography (HPLC) and enzyme immunoassays (EIA). In feces, maximum radioactivity was reached 8 h (median) post each injection, whereas maxima in urine were detected in the first samples (median 2.5 h). Metabolites excreted into feces (13.3 % ± 3.7) or urine (86.7 %) returned nearly to background levels. HPLC of fecal extracts showed minor variation between individuals and sexes. In hair, small amounts of radioactivity were present. However, two EIAs detected large amounts of unlabeled GCM, including high levels at the position of the cortisol standard; radioactivity was absent in this fraction, demonstrating that 3H-cortisol was metabolized. Furthermore, large amounts of immunoreactivity coinciding with a radioactive peak at the elution position of cortisone were found. These results show for the first time that only small amounts of systemically administered radioactive glucocorticoids are deposited in hair of guinea pigs, while measurement of large amounts of unlabeled GCM strongly suggests local production of glucocorticoids in hair follicles.

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Abbreviations

ACTH:

Adrenocorticotropic hormone

CRH:

Corticotropin releasing hormone

GC:

Glucocorticoids

GCM:

Glucocorticoid metabolites

EIA:

Enzyme immunoassay

HPA axis:

Hypothalamic–pituitary–adrenal axis

MSH:

Melanocyte-stimulating hormone

POMC:

Proopiomelanocortin

RP-HPLC:

Reversed-phase high-performance liquid chromatography

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Acknowledgments

The authors want to thank Alexandra Kuchar and Monika Höring for performing the enzyme immunoassays and Erich Möstl and Klaus Militzer for stimulating discussions throughout this study. We kindly appreciate the help of Frank Künzel with the acquisition of the application technique and the Institute for Animal Nutrition for providing the animal research facility. M. Lepschy was sponsored by the Austrian Science Fund (FWF: P-21607-B11). H. Schöpper was funded by the PhD Initiative Program BIOREC of the University of Veterinary Medicine Vienna.

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Authors and Affiliations

  1. Department for Farm Animals and Veterinary Public Health, Institute of Animal Husbandry and Animal Welfare, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210, Vienna, Austria

    Karin Keckeis & Josef Troxler

  2. Department of Biomedical Sciences, Institute of Chemistry and Biochemistry, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210, Vienna, Austria

    Michael Lepschy, Lukas Moser & Rupert Palme

  3. Department for Integrative Biology and Evolution, Research Institute of Wildlife Ecology, University of Veterinary Medicine Vienna, Savoyenstrasse 1, 1160, Vienna, Austria

    Hanna Schöpper

  4. Department of Pathobiology, Institute of Anatomy, Histology and Embryology, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210, Vienna, Austria

    Hanna Schöpper

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  1. Karin Keckeis
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  2. Michael Lepschy
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  3. Hanna Schöpper
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  4. Lukas Moser
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  5. Josef Troxler
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  6. Rupert Palme
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Correspondence to Karin Keckeis.

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K. Keckeis and M. Lepschy contributed equally to this work.

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Keckeis, K., Lepschy, M., Schöpper, H. et al. Hair cortisol: a parameter of chronic stress? Insights from a radiometabolism study in guinea pigs. J Comp Physiol B 182, 985–996 (2012). https://doi.org/10.1007/s00360-012-0674-7

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