Development and Validation of an Enzyme Immunoassay for Fecal Dehydroepiandrosterone Sulfate in Japanese Macaques (Macaca fuscata)
Measuring hormonal profiles is important in monitoring stress, physical fitness, and reproductive status in primates. Noninvasive methods have been used to measure several steroid hormones in primates without causing them stress. However, few studies have used feces or urine to measure dehydroepiandrosterone sulfate (DHEAS), an important precursor of sex steroids that has been studied as a biomarker of aging, pregnancy, and stress in humans and nonhuman primates. We developed an enzyme immunoassay to detect DHEAS in the feces of Japanese macaques (Macaca fuscata). Our subjects included eight singly housed Japanese macaques. To validate the assay, we administrated oral DHEA to one male and one female macaque, collected their feces, and measured DHEAS levels over time. Given that DHEAS is related to gonadal steroids and the stress response, we also measured DHEAS concentrations in response to adrenal (adrenocorticotropic hormone [ACTH]) and gonadal (human chorionic gonadotropin [hCG]) stimulation. Our assay successfully detected DHEAS in Japanese macaque feces, and levels of DHEAS were associated with the amount of DHEA ingested. Parallelism and accuracy tests revealed that fecal extracts were reliable measures of DHEAS. Neither ACTH nor hCG challenge appeared to affect DHEAS levels. The method we describe is less expensive than that using the commercially available kits and is applicable to investigations involving aging, stress, and reproduction in Japanese macaques.
KeywordsAging hormone Fecal steroids Noninvasive monitoring Stress hormones
We thank Mr. Akihisa Kaneko, Ms. Mayumi Morimoto, and all support staff from the Center for Human Evolution Modeling Research at the Primate Research Institute for providing useful information and assistance during the experimental procedures. We would like to express our gratitude to the journal editor and reviewers for the English language suggestions and for the very useful comments to improve this manuscript. The study was funded by the Primate Research Institute, the Leading Program in Primatology and Wildlife Science (PWS), a grant-in-aid from the Japan Society for the Promotion of Science (JSPS) no. 16 J00399, and a scholarship to R. S. C. Takeshita by the Nippon Foundation.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that there is no conflict of interest regarding the publication of this article.
- Alesci S, Bornstein S. (2001). Intraadrenal mechanisms of DHEA regulation: A hypothesis for adrenopause. Experimental and Clinical Endocrinology & Diabetes 109(2): 75–82.Google Scholar
- Baulieu, E. E., Thomas, G., Legrain, S., Lahlou, N., Roger, M., Debuire, B., Faucounau, V., Girard, L., Hervy, M. P., Latour, F., Leaud, M. C., Mokrane, A., Pitti-Ferrandi, H., Trivalle, C., de Lacharriere, O., Nouveau, S., Rakoto-Arison, B., Souberbielle, J. C., Raison, J., le Bouc, Y., Raynaud, A., Girerd, X., & Forette, F. (2000). Dehydroepiandrosterone (DHEA), DHEA sulfate, and aging: Contribution of the DHEAge study to a sociobiomedical issue. Proceedings of the National Academy of Sciences of the USA, 97(8), 4279–4284.CrossRefPubMedPubMedCentralGoogle Scholar
- Goncharova, N. D., Vengerin, A. A., & Chigarova, O. A. (2012). Repeated moderate stress stimulates the production of dehydroepiandrosterone sulfate (DHEAS) and reduces corticosteroid imbalance in old Macaca mulatta. Bulletin of Experimental Biology and Medicine, 153(5), 750–753.CrossRefPubMedGoogle Scholar
- Hammer, F., Subtil, S., Lux, P., Maser-Gluth, C., Stewart, P. M., Allolio, B., & Arlt, W. (2005). No evidence for hepatic conversion of dehydroepiandrosterone (DHEA) sulfate to DHEA: In vivo and in vitro studies. The Journal of Clinical Endocrinology & Metabolism, 90(6), 3600–3605.CrossRefGoogle Scholar
- Herrera-Justiniano, E., Galvez, M., Aznar, A., Gomez, S., Sendon, P., Zurita, A. R., Malagon, C. M., & Aznar, R. A. (1979). Changes in the plasma levels of androstenedione, dehydroepiandrosterone and cortisol after stimulation with ACTH and hCG and suppression with dexamethasone during male puberty. Acta Endocrinologica, 90(1), 113–121.PubMedGoogle Scholar
- Hu, Y., Cardounel, A., Gursoy, E., Anderson, P., & Kalimi, M. (2000). Anti-stress effects of dehydroepiandrosterone: Protection of rats against repeated immobilization stress-induced weight loss, glucocorticoid receptor production, and lipid peroxidation. Biochemical Pharmacology, 59(7), 753–762.CrossRefPubMedGoogle Scholar
- Meusy-Dessolle, N., & Dang, D. C. (1985). Plasma concentrations of testosterone, dihydrotestosterone, delta 4-androstenedione, dehydroepiandrosterone and oestradiol-17 beta in the crab-eating monkey (Macaca fascicularis) from birth to adulthood. Journal of Reproduction and Fertility, 74(2), 347–359.CrossRefPubMedGoogle Scholar
- Mobbs, C. V. (1998). Dehydroepiandrosterone and aging. In C. V. Mobbs, P. R. Hof (Eds.), Functional endocrinology of aging (Vol. 29, pp. 217–227). Basel: Karger Publishers.Google Scholar
- Möhle, U., Heistermann, M., Palme, R., & Hodges, J. K. (2002). Characterization of urinary and fecal metabolites of testosterone and their measurement for assessing gonadal endocrine function in male nonhuman primates. General and Comparative Endocrinology, 129(3), 135–145.CrossRefPubMedGoogle Scholar
- Monfort, S. L. (2003). Non-invasive endocrine measures of reproduction and stress in wild populations. In W. Holt, A. Pickard, J. Rodger, D. Wildt (Eds.), Reproductive science and integrated conservation conservation biology (Vol. 8, pp. 147–165). Cambridge: Cambridge University Press.Google Scholar
- Moran, F. M., Chen, J., Gee, N. A., Lohstroh, P. N., & Lasley, B. L. (2013). Dehydroepiandrosterone sulfate levels reflect endogenous luteinizing hormone production and response to human chorionic gonadotropin challenge in older female macaque (Macaca fascicularis). Menopause, 20(3), 329–335.PubMedPubMedCentralGoogle Scholar
- Takeshita, R. S. C., Bercovitch, F. B., Huffman, M. A., Mouri, K., Garcia, C., Rigaill, L., & Shimizu, K. (2014). Environmental, biological, and social factors influencing fecal adrenal steroid concentrations in female Japanese macaques (Macaca fuscata). American Journal of Primatology, 76(11), 1084–1093.CrossRefPubMedGoogle Scholar
- Takeshita RSC, Bercovitch FB, Kinoshita K, Huffman MA (2018). Beneficial effect of hot spring bathing on stress levels in Japanese macaques. Primates. https://doi.org/10.1007/s10329-018-0655-x.