Osmoregulatory role of vasotocinergic and isotocinergic systems in the gilthead sea bream (Sparus aurata L).

Gilthead sea bream, Sparus aurata L., is an important fish species for the Mediterranean aquaculture and is considered a good model for studying the osmoregulatory process, due to its capacity to cope with great changes in environmental salinity (5-60‰). Our group studied the osmoregulatory role of different endocrine systems in this species, focusing on the vasotocinergic and isotocinergic systems over several years. For this purpose, the cDNAs coding for pro-vasotocin (pro-vt), pro-isotocin (pro-it), two arginine vasotocin (AVT) receptors (avtr v1a2- and v2-types) and one IT receptor (itr) were cloned. Acclimation to different environmental salinities induced a direct lineal relationship between plasma AVT levels and salinity, with no changes in plasma IT values. In addition, higher values in vasotocinergic, isotocinergic and stress pathways (pro-vt and pro-it gene expression, AVT and IT storage and plasma cortisol levels) in both hypo- and/or hyper-osmotic transfers, suggest an interaction between cortisol and AVT/IT pathways. Moreover, gene expression of specific receptors, as well as the use of different in vitro techniques, demonstrated an important osmoregulatory orchestration in different organs. In addition, individuals intraperitoneally injected with AVT and transferred to different environmental salinities enhanced plasma cortisol levels and/or gill Na+, K+-ATPase activity. These effects could be related to the energy repartitioning process occurring during osmotic adaptation of S. aurata to extreme environmental salinities, which could be mediated not only by plasma cortisol but also by AVT. Finally, our results indicated a very important role of the vasotocinergic and/or isotocinergic systems in both osmoregulatory and non-osmoregulatory organs.

Unraveling vasotocinergic, isotocinergic and stress pathways after food deprivation and high stocking density in the gilthead sea bream.

The influence of chronic stress, induced by food deprivation (FD) and/or high stocking density (HSD), was assessed on stress, vasotocinergic and isotocinergic pathways of the gilthead sea bream (Sparus aurata). Fish were randomly assigned to one of the following treatments: (1) fed at low stocking density (LSD-F; 5kg·m-3); (2) fed at high stocking density (HSD-F, 40kg·m-3); (3) food-deprived at LSD (LSD-FD); and (4) food-deprived at HSD (HSD-FD). After 21days, samples from plasma, liver, hypothalamus, pituitary and head-kidney were collected. Both stressors (FD and HSD) induced a chronic stress situation, as indicated by the elevated cortisol levels, the enhancement in corticotrophin releasing hormone (crh) expression and the down-regulation in corticotrophin releasing hormone binding protein (crhbp) expression. Changes in plasma and liver metabolites confirmed a metabolic adjustment to cope with energy demand imposed by stressors. Changes in avt and it gene expression, as well as in their specific receptors (avtrv1a, avtrv2 and itr) at central (hypothalamus and pituitary) and peripheral (liver and head-kidney) levels, showed that vasotocinergic and isotocinergic pathways are involved in physiological changes induced by FD or HSD, suggesting that different stressors are handled through different stress pathways in S. aurata.

High Stocking Density and Food Deprivation Increase Brain Monoaminergic Activity in Gilthead Sea Bream (Sparus aurata).

In teleosts, brain monoamines (dopamine and serotonin) participate in the early response to different acute stressors. However, little is known regarding their role during chronic stress. In a 2 × 2 factorial design, the influence of a high stocking density (HSD) and/or food deprivation (FD) on the brain monoaminergic activity in gilthead sea bream (Sparus aurata) was evaluated. Following a 21-day experimental design, samples from the plasma and brain regions (telencephalon, hypothalamus, and optic tectum) were collected. The dopamine (DA), serotonin (5HT), and their main metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC) and 5 hydroxyindoleacetic acid (5HIAA), contents were HPLC-assessed in brain tissues, and the ratios DOPAC/DA and 5HIAA/5HT were calculated as indicators of enhanced monoaminergic activity. The plasma levels of cortisol and catecholamine were also evaluated. The cortisol levels increased in fish exposed to HSD and normally fed but, also, in all FD groups, whereas the NA levels decreased in LSD-FD animals. Within the brain, the dopaminergic and serotonergic activities in telencephalon and hypothalamus increased in fish subjected to HSD and in the telencephalon of LSD-FD fish. While DA (hypothalamus) and 5HT (telencephalon) increased in the animals submitted to a HSD, food-deprived fish did not show such an increase. Taken together, our results supported the hypothesis of brain monoaminergic activity participating in maintaining and orchestrating the endocrine response to chronic stress in fish.

Aroclor 1254 inhibits vasotocinergic pathways related to osmoregulatory and stress functions in the gilthead sea bream (Sparus aurata, Linnaeus 1758).

The present study assesses the response of vasotocinergic system in the gilthead sea bream (Sparus aurata) after administering two doses of the polychlorinated biphenyl Aroclor 1254 (15 or 50 µg g-1 fresh body mass). Seven days post-administration, eight fish of each experimental group were sampled, and the remaining animals were challenged with a hyperosmotic stress by being transferred from seawater (36 ppt) to high salinity water (55 ppt) and being sampled 3 days post-transfer. Aroclor 1254 affected gene expression of avt, together with Avt concentrations in pituitary and plasma, inhibiting the stimulation observed in vasotocinergic system after hyperosmotic challenge. This was noted by the accumulation of Avt at hypophyseal level as well as by its undetectable values in plasma. Hyperosmotic transfer significantly changed branchial avtrv1a, avtrv2, atp1a and cftr mRNA expression levels in control fish, while in Aroclor 1254-treated fish they remained mostly unchanged. This desensitization also occurred for avtrs in hypothalamus, caudal kidney and liver. In addition, an enhancement in plasma cortisol concentration, together with the orchestration of several players of the Hypothalamic-Pituitary-Interrenal axis (crh, crhbp, trh, star), was also observed mostly at the highest dose used (50 µg g-1 body mass), affecting plasma and hepatic metabolites. Our results demonstrated that Aroclor 1254 compromises the hypoosmoregulatory function of vasotocinergic system in S. aurata, also inducing a concomitant stress response. In summary, this study demonstrates that Aroclor 1254 can be considered an important endocrine disruptor in relation with the correct arrangement of vasotocinergic, metabolic and stress pathways after their stimulation by transfer to hyperosmotic environments.

The effect of starvation and re-feeding on vasotocinergic and isotocinergic pathways in immature gilthead sea bream (Sparus aurata).

This study describes the responses of the vasotocinergic and isotocinergic systems to food deprivation and re-feeding processes in immature gilthead sea bream (Sparus aurata). The animals were subjected to the following experimental treatments: (1) normal feeding (control), (2) food deprivation for 21 days; and (3) re-feeding for 7 days, beginning 14 days after starvation. The animals were sampled at 0, 7, 14 and 21 days from the beginning of the trial. The pituitary and plasma arginine vasotocin (AVT) and isotocin (IT) levels and the hypothalamic pro-vasotocin and pro-isotocin mRNA expression levels were measured. In addition, the mRNA levels of three receptors, avtr v1, avtr v2 and itr, were analyzed in target organs associated with (1) the integration and control of different physiological pathways related to stress and food intake (i.e., the hypothalamus), (2) hormonal release into the bloodstream (i.e., the pituitary), and (3) metabolism and its control (i.e., the liver). The metabolic parameters in the liver were also determined. The hepatosomatic index decreased, and hepatic metabolites were mobilized beginning in the early stages of starvation. Moreover, an over-compensation of these parameters occurred when the fish were re-fed after starvation. In terms of the vasotocinergic and isotocinergic systems, feed restriction induced a clear time-dependent regulation among metabolic organization, stress regulation and orexigenic processes in the mature hormone concentration and pro-peptide and receptor mRNA expression. Our results reveal the important role of the AVT/IT endocrine systems in the orchestration of fish physiology during starvation and re-feeding and indicate their involvement in both central and peripheral organs.