The objectives of this study were to characterize rainbow trout (Oncorhynchus mykiss) corticotropin-releasing factor (CRF)-binding protein (CRF-BP) cDNA and to examine the variations in CRF-BP and CRF mRNA levels in response to different intensities of stress. Trout were physically disturbed by a single or three consecutive periods of chasing until exhaustion followed by 2 h of recovery. The pituitary CRF-BP and preoptic area CRF1 mRNA contents were significantly increased only after repeated chasing events. Physical disturbance increased plasma cortisol levels with the largest change occurring in the group of trout that were exposed to repeated chasing events. Trout were also individually isolated in 120 l tanks or confined to 1.5 l boxes for 4, 24 or 72 h. CRF-BP mRNA levels in confined fish were greater than those of isolated fish at 72 h although there were no differences compared with the control group. CRF1 mRNA levels in the preoptic area were greater and remained elevated for a longer period in confined compared with isolated trout. Isolation led to a transient increase in plasma cortisol levels, but the higher cortisol values developed in the confined fish suggest that this treatment was more stressful than isolation. These results demonstrate that the intensity and duration of stress are important factors regulating CRF and CRF-BP mRNA levels in rainbow trout. We hypothesize that pituitary CRF-BP is involved in regulating the activity of the stress axis, possibly by reducing access to CRF1 receptors in the corticotropes.

ABSTRACT

Pyroglutamylglutamylprolineamide (pGlu-Glu-ProNH₂) is a tripeptide with structural and immunological similarities to thyrotrophin-releasing hormone (TRH; pGlu-His-ProNH₂). Since TRH stimulates GH secretion in domestic fowl, the possibility that pGlu-Glu-ProNH₂ may also provoke GH release was investigated. Unlike TRH, pGlu-Glu-ProNH₂ alone had no effect on GH release from incubated chicken pituitary glands and did not down-regulate pituitary TRH receptors. However, pGlu-Glu-ProNH₂ suppressed TRH-induced GH release from pituitary glands incubated in vitro and competitively displaced [³H]methyl³-histidine²-TRH from pituitary membranes. Systemic injections of pGlu-Glu-ProNH₂ had no significant effect on basal GH concentrations in conscious birds, but promptly lowered circulating GH levels in sodiumpentobarbitone anaesthetized fowl. Submaximal GH responses of conscious and anaesthetized birds to systemic TRH challenge were, however, potentiated by prior or concomitant administration of pGlu-Glu-ProNH₂. These results demonstrate, for the first time, that pGlu-Glu-ProNH₂ has biological activity, with inhibitory and stimulatory actions within the avian hypothalamo-pituitary axis. These results indicate that pGlu-Glu-ProNH₂ may act as a TRH receptor antagonist within this axis.

Journal of Endocrinology (1993) 138, 137–147

ABSTRACT

The effects of acute i.v. administration of gonadotrophin-releasing hormone (GnRH; 0·1 μg/kg), morphine (3 mg/kg) and/or naloxone (0·5 mg/kg) on LH and FSH secretion was evaluated in young male pigs (approximately 6 weeks old) with venous brachiocephalic cannulae. The effects of morphine and/or naloxone treatments on prolactin and GH were also evaluated. The influence of morphine on hypophysial hormone secretion was also examined 2 days after castration. Animals treated with morphine and/or naloxone were compared with saline-injected control animals. Injection of GnRH induced 400 and 50% increases in LH and FSH respectively. Morphine and/or naloxone did not influence LH secretion in intact or castrated animals. Morphine suppressed (P < 0·01) FSH levels 40–60 min after injection whereas naloxone had no effect. Castration eliminated morphine-induced suppression of FSH. Injection of morphine followed by naloxone resulted in acutely raised (P < 0·05) FSH concentrations. Morphine induced a threefold increase (P < 0·01) in prolactin within 30 min of injection and naloxone inhibited the effect of morphine. Levels of GH were increased (P < 0·01) 20 min after morphine treatment and this increase was delayed when naloxone was given immediately after morphine. Naloxone alone did not affect prolactin or GH secretion. Castration caused increases in LH (P < 0·05) and FSH (P < 0·01), did not influence prolactin or GH, and reduced plasma testosterone to undetectable (< 1·0 nmol/l) levels. These results suggest that in young male pigs the hypothalamic-hypophysial axis is responsive to GnRH and gonadal negative feedback. The opiate/LH pathway appears to be non-functional or incomplete, while the opiate/FSH pathway seems to be active. Morphine stimulated the release of prolactin probably via a naloxone-sensitive opiate receptor.

J. Endocr. (1988) 119, 501–508