Influence of environmental colour and diurnal phase on MCH gene expression in the trout.

Melanin-concentrating hormone (MCH) gene expression in the brain of rainbow trout, reared and maintained in either pale or black-coloured tanks, was studied using in situ hybridization histochemistry. MCH transcripts were most prevalent in the magnocellular neurones of the nucleus lateralis tuberis (NLT), which project to the pituitary gland. They were also present, although at much lower levels, in dorsally projecting parvocellular neurones, sited more posteriorly above the lateral ventricular recess (LVR). In the NLT the most intense hybridization signal was seen over the pituitary stalk; above the LVR, the most active neurones were located caudally. In both the NLT and above the LVR, MCH hybridization signal was 4-fold stronger in white-reared fish than in black-reared fish. There was also a marked diurnal variation in MCH expression in both sites, with high levels at 16.00 h and lower levels at 04.00 h. The results show that gene activity in both hormonal (NLT) and neuromodulator/neurotransmitter (LVR) MCH neurones is induced by pale environmental colour and that MCH gene activity is subject to pronounced diurnal variation.

Neuropeptide Y stimulates food consumption through multiple receptors in goldfish.

In this study, the acute effects of brain intracerebroventricular (icv) injections of mammalian neuropeptide Y (NPY) Y1 ([31Leu,34Pro]NPY), Y2 (NPY2-36) and Y5 ([D-32Trp]NPY) receptor subtype agonists on food intake in goldfish were examined. Icv injection of Y1 and Y5 receptor agonists in dosages of 1 and 5 ng/g exhibited dose-dependent effects on food intake; however, higher dosages of both receptor subtypes had desensitising effects on food intake, and caused a decrease in food intake in comparison to the lower dosages. At 10 and 20 ng/g, Y1 receptor agonist-treated fish exhibited feeding significantly lower than intact and saline-injected fish; fish treated with the same dosages of Y5 agonist exhibited food intake similar to intact and saline-injected fish. Y2 agonist had no effects on food intake. Co-icv administration of Y1 and Y5 agonists in dosages of 1 and 5 ng/g caused enhanced food intake that was additive of the individual doses alone. However, desensitising one receptor did not influence the responsiveness of the other. Co-icv injection of Y1 receptor agonist in desensitizing high dosages (10 and 15 ng/g) with Y5 receptor agonist in lower doses (1 and 5 ng/g, respectively) or vice versa, resulted in a food intake similar to the dosages of Y1 and Y5 receptor agonists at 1 and 5 ng/g given alone. Overall, this study demonstrates that NPY acts centrally through Y1 and Y5 receptors to stimulate food intake in goldfish. The Y1 and Y5 receptors appear to act independently in the stimulation of food intake in goldfish.

Stimulation of macrophage phagocytosis and lymphocyte count by exogenous prolactin administration in silver sea bream (Sparus sarba) adapted to hyper- and hypo-osmotic salinities.

Juvenile silver sea bream (Sparus sarba) were adapted to hyper- (33 ppt) and hypo-osmotic (6 ppt) salinities for 3 weeks and injected daily with ovine prolactin (1 microg/g body weight i.p.) during the last 7 days of the adaptation period. Fish injected with prolactin exhibited significant increases in percent phagocytosis and phagocytic index of both pronephric and splenic macrophages regardless of salinity. Prolactin injection also resulted in elevated blood lymphocyte counts in both hyper- and hypo-osmotically adapted sea bream. The present results provide further evidence for the existence of a neuroendocrine-immune link in teleosts.

Effects of food deprivation and refeeding on neuropeptide Y (NPY) mRNA levels in goldfish.

In mammals, NPY is a key factor in the regulation of feeding behavior. In the present study, the effects of refeeding for 1-3 h in 72-75-h food deprived (FD) goldfish on the levels of NPY mRNA in telencephalon-preoptic (TEL-POA), hypothalamus (HYP) and optic tectum-thalamus (OT-THAL) were examined, using Northern blot analysis. Goldfish FD for 72 h exhibited a significant increase in NPY mRNA levels in all brain regions. At 1 h after 72-h FD (73-h FD), NPY mRNA was significantly increased in TEL-POA and OT-THAL, but remained the same as 72-h FD fish in HYP. At 3 h after 72-h FD (75 h), all brain regions exhibited a significant increase in NPY mRNA levels. However, subsequent refeeding for 1-3 h rapidly and completely reversed the effects of FD in all brain regions, reaching fed levels within 1-3 h of refeeding. Serum GH levels were highest in 72-h FD fish, but decreased significantly over 1-3 h after 72-h FD; whereas, refeeding reversed the increase in serum GH levels only at 3 h after refeeding. Taken together, these results further support that NPY is a physiological brain transducer involved in the regulation of daily appetite and feeding in goldfish.

Evidence that cortisol may protect against the immediate effects of stress on circulating leukocytes in the trout.

Rainbow trout stressed by an intraperitonal injection of saline displayed reduced phagocytic activity of their spleen and head-kidney macrophages within 3 hr. Phagocytic activity was similarly depressed by injecting noradrenalin, but was maintained in fish injected with the adrenergic blocking agent phentolamine, suggesting that endogenous catecholamines are involved in this stress response. Since stress may increase the number of circulating granulocytes, it is proposed that noradrenalin, released during stress, causes the liberation of active macrophages from the lymphocytic tissue, the remaining macrophages therefore showing a lowered phagocytic index. Cortisol injection, like phentolamine, prevented the depressive effect of stress on the phagocytic index but did not antagonize the depressive effect of exogenous noradrenalin. It is suggested that the stress-induced release of endogenous catecholamines may be prevented by cortisol. Injection stress caused a decline in the number of circulating lymphocytes/thrombocytes, indicating their retrafficking into some other tissue. This was opposed by cortisol and by high doses of noradrenalin. It is proposed that cortisol or noradrenalin may oppose, directly or indirectly, the expression of adhesion molecules which are normally induced after stress.

The effect of various stresses, corticosteroids and adrenergic agents on phagocytosis in the rainbow trout Oncorhynchus mykiss.

The effect of acute and chronic stress on the phagocytic activity of putative macrophages from the rainbow trout. Oncorhynchus mykiss has been assessed, using an in vitro phagocytic index, in which the average number of engulfed yeast cells in a population of phagocytes is determined. An injection stress given under light anaesthesia, or a longer noise stress combined with confinement, both significantly reduced, within 3 h, the level of phagocytic activity of macrophages from the spleen and pronephros. Daily injection stress over six days had a lesser effect on the proportion of phagocytically active cells even though plasma cortisol levels were equally raised. Daily dexamethasone injection depressed the proportion of phagocytically active cells more than saline injection. In these in vivo experiments, it was not possible to determine whether stress and steroids depressed the phagocytic activity of individual macrophages or caused the active macrophages to migrate out of the spleen and pronephros. Administration of cortisol (200 nM) to trout macrophages in vitro failed to depress phagocytic activity within a 3h period but both α- and ß-adrenergic agonists (10 µM) were usually depressive. It is proposed that the autonomic nervous system may be an early regulator of macrophage phagocytosis following stress and that corticosteroids only exert their suppressive effect on macrophage activity in the longer term.

Regulation of food intake by neuropeptide Y in goldfish.

In mammals, neuropeptide Y (NPY) is a potent orexigenic factor. In the present study, third brain ventricle (intracerebroventricular) injection of goldfish NPY (gNPY) caused a dose-dependent increase in food intake in goldfish, and intracerebroventricular administration of NPY Y1-receptor antagonist BIBP-3226 decreased food intake; the actions of gNPY were blocked by simultaneous injection of BIBP-3226. Goldfish maintained on a daily scheduled feeding regimen display an increase in NPY mRNA levels in the telencephalon-preoptic area and hypothalamus shortly before feeding; however, a decrease occured in optic tectum-thalamus. In both fed and unfed fish, brain NPY mRNA levels decreased after scheduled feeding. Restriction in daily food ration intake for 1 wk or food deprivation for 72 h resulted in increased brain NPY mRNA levels. Results from these studies demonstrate that NPY is a physiological brain signal involved in feeding behavior in goldfish, mediating its effects, at least in part, through Y1-like receptors in the brain.