Ontogeny of corticotropin-releasing factor and of hypothalamic-pituitary-interrenal axis responsiveness to stress in tilapia (Oreochromis mossambicus; Teleostei).

The ontogeny of the corticotropin-releasing factor (CRF) system and of the ability of the hypothalamic-pituitary-interrenal (HPI) axis to respond to stressors (capture or confinement), or to cortisol treatment was investigated in tilapia (Oreochromis mossambicus). In 2 days post hatching (dph) larvae, the first developmental stage used for immunohistochemistry, CRF-immunoreactivity (ir) was observed in the nucleus preopticus (npo), and in two hypothalamic nuclei (nlt and nrl). In this stage, CRF- and AVT-ir was found in the neural part of the pituitary, and endocrine cells in the pars distalis and pars intermedia contained POMC-derived peptides. In the ventral telencephalon, CRF-ir cells were first observed 5 dph, whereas projections from these cells into the anterior part of the latero-dorsal telencephalon (Dla) from 7 dph onwards. CRF, ACTH, alpha-MSH, and cortisol were quantified by radioimmunoassays in homogenates of the anterior-cranial region of the larvae containing brain, pituitary, and headkidneys. CRF contents increased from 43 +/- 3 to 1070 +/- 70 pg/larvae between 5 and 110 dph. Larvae of age 5, 12, 24, and 42 dph were captured sequentially from a group. All life stages were able to rapidly increase their cortisol content in response to this stressor (ANOVA: P < 0.001). Overall, the developmental stage affected cortisol content (ANOVA: P < 0.001), but developmental stage did not influence the cortisol reaction to stress (ANOVA: P > 0.162). Whole brain CRF content did not change during the 20 min stress period and the relationship between CRF-producing neurons and the initial HPI stress response in early life stages remains to be established. Cortisol feeding of 18 and 29 dph larvae for periods ranging from 2 to 24 days resulted in elevations of the CRF content (P < 0.003) in comparison to controls. In 18 dph larvae cortisol feeding abolished the cortisol response to capture stress as observed in control fed larvae (P < 0.008). We propose that cortisol induced upregulation of CRF takes place in the telencephalon and is restricted to a time period during larval development, characterised by the absence of glucocortoid receptor (GR) expression in the telencephalic Dm region in these larvae. Finally, the stress response to 24 h confinement was compared between saltwater adapted and freshwater adapted juveniles (age 77 dph). Confinement stress (24 h) affected cortisol and CRF content (ANOVA: P < 0.001, P < 0.008, respectively), but not ACTH content. Interactions were observed between salinity and confinement regarding cortisol and alpha-MSH contents (ANOVA: P < 0.02), but not regarding CRF and ACTH contents. The increase in cortisol levels induced by confinement was remarkably high in freshwater adapted larvae (five times higher than in saltwater adapted larvae). Regarding the cortisol response it is concluded that during and after the period of mouth breeding tilapia larvae respond to capture stress in a similar fashion (onset and height) as adults. Previously, we reported that the initial plasma cortisol response to capture stress in adult tilapia occurred independently from changes in plasma ACTH levels. The current finding that also brain CRF contents do not alter during the initial cortisol response in larvae further indicates that the initial cortisol response in this species may be regulated independently from CRF and ACTH.