Chronic and acute stress responses in Senegalese sole (Solea senegalensis): the involvement of cortisol, CRH and CRH-BP.

The hypothalamus-pituitary-interrenal (HPI) axis is pivotal in the endocrine stress response of fish. Hypothalamic corticotropin-releasing hormone (CRH) initiates the endocrine stress response and stimulates the release of adrenocorticotropic hormone (ACTH) from the pituitary pars distalis, which in turn activates cortisol production and release by the interrenal cells of the head kidney. CRH activity depends on the levels of a specific CRH binding protein (CRH-BP). We have characterized the cDNAs coding for CRH and CRH-BP in Senegalese sole (Solea senegalensis) and investigated their mRNA expression in juveniles that were submitted to a protocol that involved exposure to a chronic stressor (viz. increased cultivation densities) followed by an acute stressor (viz. transfer to increased ambient salinity). Juveniles were cultivated at three densities (1.9, 4.7 and 9.8 kg/m(2)) for 33 days, and then exposed to an osmotic challenge that involved transfer from seawater (39‰ salinity, SW) to hypersaline seawater (55‰, HSW). The highest density imposed stress as indicated by elevated cortisol levels and CRH mRNA expression compared to fish stocked at low density. Fish kept at high density differentially responded to a posterior transfer to HSW; no cortisol or CRH response was seen, but osmoregulatory and metabolic parameters were affected. No differences in CRH-BP mRNA expression levels were found at different stocking densities; transfer to HSW enhanced expression in both low and high density stocked animals, suggesting that CRH-BP acts as a modulator of the acute stress response, not so of the chronic stress response. We conclude that stocking of Senegalese sole at high density is a stressful condition that may compromise the capacity to cope with subsequent stressors.