Short communication: Skin melatonin and cortisol responses to water acidification and basification within the optimal pH range in three-spined sticklebacks.

Fish skin functions not only as a passive protective barrier but also as an active site of key physiological processes, including a local stress response system. In fish, this system involves the hormones cortisol and melatonin (Mel), which contribute to counteracting environmental stressors and maintaining homeostasis. In this study, we examined the sensitivity of both components of the cutaneous stress response system (CSRS) in three-spined sticklebacks (Gasterosteus aculeatus) exposed to acidic (pH = 6.54) and basic (pH = 8.70) water conditions, representing the boundary values of the species' optimal pH range, under either rapid or gradual pH change regimes. Water pH in the aquaria was precisely controlled using a custom-designed gas-exchange system regulating dissolved CO2 levels. Mel concentrations were measured in the skin, brain and eyeball, while cortisol was determined in the skin. Samples were collected during the day. Skin Mel levels were significantly higher under acidification than under basification (P = 0.036; rapid change regime), whereas cortisol remained stable across all conditions. Ocular Mel levels were not affected by treatments. Brain Mel concentrations were generally very low but tended to be slightly higher under basification than under acidification in both regimes (P = 0.05, borderline significance). The marked differences in skin Mel levels between acidic and basic pH water conditions, accompanied by stable cortisol concentrations, indicates that cutaneous Mel, but not cortisol, is highly sensitive to subtle water pH fluctuations even within the species' optimal range.