The adrenal stress response involves distinct dynamics of both cortisol and corticosterone in the axolotl salamander.

The axolotl is a popular model organism in regenerative biology owing to its ability to regenerate amputated limbs and internal organs. The role of injury-derived signals in initiating the regenerative response is still not well understood, but the potential involvement of the stress response is of interest, as injury and stress are temporally linked. The dominant glucocorticoid response to stress varies among species, with corticosterone generally considered dominant in most amphibians, whereas cortisol predominates in others. Here we characterize the adrenal stress response in the axolotl and describe methods to measure axolotl stress hormones to facilitate their inclusion in future research involving axolotl development and regeneration. We describe an intricate and unexpected axolotl stress response that involves cortisol and corticosterone, each being dominant under different conditions. Corticosterone is preferably activated by the classical hypothalamus-pituitary-interrenal axis pathway, with both arginine vasotocin and adrenocorticotropic hormone promoting its synthesis and release. Under manual stress and direct stimuli with acetylcholine, cortisol is more prominent, suggesting an alternative mechanism involving sympathetic nerve signaling. In response to an amputation injury, both cortisol and corticosterone are increased, with corticosterone being dominant, suggesting an injury-specific response. Finally, when administering glucocorticoids directly and measuring classical physiological effects of glucocorticoid signaling, cortisol is more potent. We propose a hypothesis that axolotls rely on cortisol as their dominant glucocorticoid, functioning in part as an extension of the catecholamine system. By contrast, corticosterone is mainly regulated classically via the hypothalamus-pituitary-interrenal axis.