High density and food deprivation affect arginine vasotocin, isotocin and melatonin in gilthead sea bream (Sparus auratus).

Arginine vasotocin (AVT) and isotocin (IT) levels in plasma and pituitary, and melatonin (MEL) levels in plasma were determined in gilthead sea bream (Sparus auratus) subjected to two different types of stress: i) high density (HD) and ii) food deprivation (NF: non-fed). Fishes were randomly assigned to one of 4 treatments that lasted for 14 days: 1) fed fish under normal low density (ND, 4 kg m(-3)); 2) non-fed (NF) fish under ND; 3) fed fish under high density (HD, 70 kg m(-3)); and 4) non-fed fish under HD. Ten fish from each tank were anaesthetized, weighed and plasma and pituitary samples were taken. Plasma and pituitary AVT and IT content were determined by HPLC, while plasma MEL was assayed by RIA. Plasma AVT and IT values were enhanced in all fish kept at high density. The response of AVT was much stronger than that of IT. The highest pituitary AVT and IT levels were shown in NF fish kept at normal density. The significantly higher plasma MEL levels were measured in fed fish kept at HD. These results suggest a role of AVT, IT and MEL in response of sea bream to a common stress factor, high density. Although food deprivation does not influence AVT and IT plasma levels, it seems to affect hypothalamic synthesis of nonapeptides. Further studies are required to elucidate the complex role of AVT, IT and MEL in the sea bream's response to different stress stimuli.

Concentration of neuropeptide Y, galanin, &bgr;-endorphin, vasoactive intestinal peptide and gonadotropin releasing hormone in the hypothalamus of gilts during the estrous cycle.

Numerous reports indicate that peptides isolated from the brain such as beta-endorphin (beta-END), neuropeptide Y (NPY), galanin (GAL) or vasoactive intestinal peptide (VIP), modulate secretion of gonadotropins and prolactin. The objective of the present experiment was to determine concentrations of NPY, GAL, beta-END, VIP and GnRH in the preoptic area (POA), medial basal hypothalamus (MBH) and pituitary stalk-median eminence (SME) during the estrous cycle in the pig. Gilts were slaughtered on Days 5, 10, 15 and 20 of the estrous cycle. Blood samples for analyses of progesterone were taken before slaughter. Neuropeptide concentrations in brain tissues were determined using RIA. The highest concentrations of all determined peptides occurred in SME. GnRH concentration in MBH was lower (p<0.05) in POA and SME on Day 20 than on Day 5. NPY concentration in POA was 5-6 times greater (p<0.05) on Days 10 and 20 than on Day 5. Similarly, concentrations of VIP in POA were greater (p<0.05) on Day 10, Day 15 and Day 20 than on Day 5. The concentration of GAL in POA was higher on Days 10 and 15 (p<0.05) than on Days 5 and 20. The concentration of GAL in SME was lowest on Day 5 and then significantly increased on Days 10, 15 and 20. In SME, concentration of beta-END increased 10 times on Days 15 and 20 when compared to Day 5 of the cycle. The correlation between concentration of GAL in the POA and MBH and progesterone concentration in the peripheral blood was positive, whereas this correlation associated with the SME was negative. These results indicate that considerable changes in various neuropeptide concentrations in different areas of the porcine hypothalamus are associated with stage of the estrous cycle and that GAL may be involved in control of the preovulatory LH surge in pigs.

Melatonin and thyroxine response to pollution in white stork nestlings (Ciconia ciconia): aspects of rhythmicity and age.

There is growing evidence that ubiquitous environmental contaminants may interfere with vertebrate endocrine systems. The selected endocrine biomarkers are used to indicate the condition of free-ranging populations of wildlife, including avian species. The aim of this study was to determine the impact of environment quality on serum thyroxine (T4) and melatonin (Mel) in white stork nestlings (Ciconia ciconia) living in different locations: small villages in natural areas surrounded by forests and crop fields, near the city and near the copper smelter. We extended our analyses to examine the hormones' day-night changes in conjunction with chicks' age. Total serum T4 and Mel was measured by RIA. T4 level, as a decisive measure of thyroid hormone productivity, was significantly lower in the nestlings exposed to pollutants from the copper smelter. Mel, as a well-known scavenger of free radicals, was elevated in the nestlings in the area near the copper smelter. This study indicates that alteration in T4 and Mel levels could be a useful marker of exposure of nestling wild storks to different toxic substances in field studies. Mel is postulated to be a susceptible defensive molecule as a protective mechanism for organisms.

Day-night specific binding of 2-[125I]iodomelatonin and melatonin content in gill, small intestine and kidney of three fish species.

Some of melatonin's (Mel) well-established physiological effects are mediated via high-affinity cell-membrane receptors belonging to the superfamily of G-protein-coupled receptors. Specific binding of ligand 2-[(125)I]iodomelatonin, using membrane preparations from osmoregulatory tissues of flounder, rainbow trout and sea bream, together with Mel concentrations in the tissues and plasma were studied. The kidney, gill and small intestine samples were collected during the day and at night. The dissociation constants (K (d)) and maximal binding densities (B (max)) were calculated for each tissue at 11:00 and 23:00 h. The binding sites with K (d) values in the tissues in the picomolar range indicated the high affinity. K (d) and B (max) values were tissue- and species-dependent. The GTP analogue [Guanosine 5'-O-(3-thiotriphosphate)] treatment significantly reduced the B (max) value, indicating that the 2-[(125)I]iodomelatonin-binding sites are probably coupled to a G-protein. No daily variations in K (d) and B (max) values were observed. These are the first studies of the presence of 2-[(125)I]iodomelatonin-binding sites in the small intestine, kidney tubule and gill of fish. The data strongly suggest new potential targets for Mel action and the influence of Mel on water/ion balance in fish. The intestine seems to be a site of Mel synthesis and/or an active accumulation of the hormone.

Arginine vasotocin, isotocin and melatonin responses following acclimation of gilthead sea bream (Sparus aurata) to different environmental salinities.

Gilthead sea bream (Sparus aurata) is a euryhaline species with a capacity to cope with demands in a wide range of salinities and thus is a perfect model-fish to study osmoregulatory responses to salinity-adaptive processes and their hormonal control. Immature sea bream acclimated to different salinities, i.e. SW (38 per thousand), LSW (5 per thousand) and HSW (55 per thousand), were kept at 18 degrees C under natural photoperiod. Arginine vasotocin (AVT) and isotocin (IT) in plasma and pituitary were determined by HPLC. Plasma melatonin (Mel) was assayed by RIA. Plasma osmolality, ion concentrations (Na(+), K(+), Ca(2+), Cl(-)) and Na(+),K(+)-ATPase activity in gill were measured. A steady increase in plasma AVT, along with increasing water salinity was observed. Pituitary IT concentration in HSW-acclimated fish was significantly higher than that in LSW group. AVT/IT secretory system of sea bream does appear to be involved in the mechanism of long-term acclimation to different salinities. The distinct roles and control mechanisms of both nonapeptides are suggested. Plasma Mel was significantly higher in LSW compared with both HSW and SW groups. Data indicate that the changes in Mel level are linked to osmoregulation. Further studies are required to elucidate a complex role of AVT, IT and Mel in sea bream osmoregulation.

Seasonal changes in brain melatonin concentration in the three-spined stickleback (Gasterosteus aculeatus): towards an endocrine calendar.

Pineal organ and its hormone melatonin (N-acetyl-5-methoxytryptamine) is likely involved in timing and synchronisation of many internal processes, such as reproduction, with annual changes in environmental cues, i.e., photoperiod and water temperature. The seasonal changes in melatonin profile in stickleback brains related to the following reproductive phases were examined, and the link between melatonin concentrations and the stages of spawning cycle was analysed. Two wild populations of sticklebacks were exposed to annual environmental changes in their natural habitats. Brains, gonads, kidneys and livers were collected over 2 years. Melatonin was measured using RIA and the indices, gonadosomatic (GSI), nephrosomatic (NSI) and hepatosomatic (HSI), were calculated. The role of melatonin, as a component of internal calendar engaged in the control of seasonal breeding in this species, is discussed. The extremely high melatonin levels observed in early spring (March) and autumn (October) seem to mark out a time frame for spawning in sticklebacks. The seasonal pattern of melatonin production and identified development stages of gonads suggests the potential inhibitory effect of the hormone on stickleback reproduction in shortening photoperiod and stimulatory effect in lengthening photoperiod.