Measurement of 11-dehydrocorticosterone in mice, rats and songbirds: Effects of age, sex and stress

• Published in: General and comparative endocrinology Vol. 281; pp. 173 - 182
• Main Authors: Hamden, Jordan E., Salehzadeh, Melody, Jalabert, Cecilia, O'Leary, Timothy P., Snyder, Jason S., Gomez-Sanchez, Celso E., Soma, Kiran K.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15.09.2019
• Subjects: 11β-Hydroxysteroid dehydrogenase; Aging - metabolism; Animals; Antibodies - metabolism; Corticosterone - analogs & derivatives; Corticosterone - blood; Corticosterone - chemistry; Cross Reactions; Development; ELISA; Female; Glucocorticoid; Glucocorticoids - chemistry; Glucocorticoids - metabolism; Immunosteroid; Male; Mice, Inbred C57BL; Rats, Long-Evans; Reference Standards; Sex Characteristics; Sex difference; Songbirds - blood; Stress, Physiological; Development; Immunosteroid; Glucocorticoid; Sex difference; 11β-Hydroxysteroid dehydrogenase; ELISA
• ISSN: 0016-6480; 1095-6840
• DOI: 10.1016/j.ygcen.2019.05.018

•We developed an immunoassay for 11-dehydrocorticosterone, an important steroid.•We validated the assay for accuracy, precision, specificity, and matrix effects.•In mice, corticosterone and 11-dehydrocorticosterone levels vary with age and tissue.•In rats, restraint increases corticosterone and 11-dehydrocorticosterone levels.•In song sparrows, restraint increases corticosterone and 11-dehydrocorticosterone levels. Glucocorticoids (GCs) are secreted into the blood by the adrenal glands and are also locally-produced by organs such as the lymphoid organs (bone marrow, thymus, and spleen). Corticosterone is the primary circulating GC in many species, including mice, rats and birds. Within lymphoid organs, corticosterone can be locally produced from the inactive metabolite, 11-dehydrocorticosterone (DHC). However, very little is known about endogenous DHC levels, and no immunoassays are currently available to measure DHC. Here, we developed an easy-to-use and inexpensive immunoassay to measure DHC that is accurate, precise, sensitive, and specific. The DHC immunoassay was validated in multiple ways, including comparison with a mass spectrometry assay. After assay validations, we demonstrated the usefulness of this immunoassay by measuring DHC (and corticosterone) in mice, rats and song sparrows. Overall, corticosterone levels were higher than DHC levels across species. In Study 1, using mice, we measured steroids in whole blood and lymphoid organs at postnatal day (PND) 5, PND23, and PND90. Corticosterone and DHC showed distinct tissue-specific patterns across development. In Studies 2 and 3, we measured circulating corticosterone and DHC in adult rats and song sparrows, before and after restraint stress. In rats and song sparrows, restraint stress rapidly increased circulating levels of both steroids. This novel DHC immunoassay revealed major changes in DHC concentrations during development and in response to stress, which have important implications for understanding GC physiology, effects of stress on immune function, and regulation of local GC levels.