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Eyestalk is considered the main neuroendocrine organ in crustaceans. Eyestalk regulates reproduction, molting, and energy metabolism by secreting several neurohormones. However, the role of eyestalk in salinity adaptation in crustaceans remains unclear. To reveal the role of eyestalk in salinity adaptation in Litopenaeus vannamei, we performed RNA-seq to compare the transcriptomic response of the eyestalk under low salinity (salinity 3) with that of the control group (salinity 25) for 8 weeks. A total of 479 mRNAs, including 150 upregulated and 329 downregulated mRNAs, were differentially expressed between the two salinity groups. The majority of the differentially expressed genes (DEGs) were enriched in biological pathways related to osmoregulation, metabolism and energy production, and oxidative stress. The most important DEGs associated with osmoregulation were CA4, ATP1A, ATP2B, ABCB1, ABCC4, PhoA, PhoB, NOS1, ACE, ANPEP, and the V-type H

Salinity is one of the main physical properties that govern the distribution of fishes across aquatic habitats. In order to maintain their body fluids near osmotic set points in the face of salinity changes, euryhaline fishes rely upon tissue-level osmotically-induced responses and systemic endocrine signaling to direct adaptive ion-transport processes in the gill and other critical osmoregulatory organs. Some euryhaline teleosts inhabit tidally influenced waters such as estuaries where salinity can vary between fresh water (FW) and seawater (SW). The physiological adaptations that underlie euryhalinity in teleosts have been traditionally identified in fish held under steady-state conditions or following unidirectional transfers between FW and SW. Far fewer studies have employed salinity regimes that simulate the tidal cycles that some euryhaline fishes may experience in their native habitats. With an emphasis on prolactin (Prl) signaling and branchial ionocytes, this mini-review contrasts the physiological responses between euryhaline fish responding to tidal versus unidirectional changes in salinity. Three patterns that emerged from studying Mozambique tilapia (Oreochromis mossambicus) subjected to tidally-changing salinities include, 1) fish can compensate for continuous and marked changes in external salinity to maintain osmoregulatory parameters within narrow ranges, 2) tilapia maintain branchial ionocyte populations in a fashion similar to SW-acclimated fish, and 3) there is a shift from systemic to local modulation of Prl signaling.

The caudal neurosecretory system (CNSS) is a part of stress response system, a neuroendocrine structure unique to fish. To gain a better understanding of the physiological roles of CNSS in fluid homeostasis, we characterized the tissue distribution of urotensin I (UI) expression in European flounder (Platichthys flesus), analyzed the effect chronic exposure to seawater (SW) or freshwater (FW), transfer from SW to FW, and reverse transfer on mRNA levels of UI, L-type Ca2+ channels and Ca-activated K+ channels transcripts in CNSS. The tissue distribution demonstrated that the CNSS is dominant sites of UI expression, and UI mRNA level in fore brain appeared greater than other non-CNSS tissues. There were no consistent differences in CNSS UI expression or urophysis UI content between SW- and FW-adapted fish in July and September. After transfer from SW to FW, at 8 h CNSS UI expression was significantly increased, but urophysis UI content was no significantly changes. At 24 h transfer from SW to FW, expression of CNSS UI was no apparent change and urophysis UI content was reduced. At 8 h and 24 h after transfer from FW to SW UI expression and urophysis UI content was no significantly effect. The expression of bursting dependent L-type Ca2+ channels and Ca-activated K+ channels in SW-adapted fish significantly decreased compared to those in FW-adapted. However, there were no differences in transfer from SW to FW or from FW to SW at 8 h and 24 h. Thus, these results suggest CNSS UI acts as a modulator in response to osmotic stress and plays important roles in the body fluid homeostasis.

Anadromous sea lamprey (Petromyzon marinus) larvae undergo a months-long true metamorphosis during which they develop seawater (SW) tolerance prior to downstream migration and SW entry. We have previously shown that intestinal Na

Non-native strains of Atlantic salmon are used in reinstatement trials where populations are extinct. Environmental cues like photoperiod and temperature are known to influence the smolting process and there is evidence of strain-, stock-or population-specific differences associated with seaward migration or smoltification. The objective of this study was to compare morphological, osmoregulatory and endocrine features between two strains, one originating froma cold and short river in Ireland (Cong) and another from a long and warm river in France (Loire-Allier), reared under Belgian conditions in order to highlight major differences in restocking adaptability. Comprehensive endocrine profiles, consistent with their interactive role of mediating changes associated with smolting,have been observed. Na+/K+ATPase activity (1.3-10.5 μmol ADP*mg prot.-1*h-1) and hormone plasma levels (e.g. 55-122 ng*mL-1of cortisol and 4.5-6.4 ng*mL-1of GH) were consistent with reported values. We observed strain-related differences of the influence of temperature and daylength on cortisol, GH and sodium plasma levels. These may be related to the respective environmental conditions prevailing in the river of origin, which have impacted the genetic background for smoltification. Using Na+/K+ATPase activity as an indicator, both strains smoltified successfully and simultaneously testifying a prevailing influence of environmental cues over genetic factors for smoltification., Non-native strains of Atlantic salmon are used in reinstatement trials where populations are extinct. Environmental cues like photoperiod and temperature are known to influence the smolting process and there is evidence of strain-, stock- or population-specific differences associated with seaward migration or smoltification. The objective of this study was to compare morphological, osmoregulatory and endorcrine features between two strains, one originating from a cold and short river in Ireland (Cong) and another from a long and warm river in France (Loire-Allier), reared under Belgian conditions in order to highlight major differences in restocking adaptability. Comprehensive endocrine profiles, consistent with their interactive role of mediating changes associated with smolting, have been observed. Na+/K+ATPase activity (1.3–10.5 µmol ADP∗mg prot.−1∗h−1) and hormone plasma levels (e.g. 55–122 ng∗mL−1 of cortisol and 4.5–6.4 ng∗mL−1 of GH) were consistent with reported values. We observed strain-related differences of the influence of temperature and daylength on cortisol, GH and sodium plasma levels. These may be related to the respective environmental conditions prevailing in the river of origin, which have impacted the genetic background for smoltification. Using Na+/K+ATPase activity as an indicator, both strains smoltified successfully and simultaneously testifying a prevailing influence of environmental cues over genetic factors for smoltification.

Osmoregulation in vertebrates is largely controlled by the neuroendocrine system. Prolactin (PRL) is critical for the survival of euryhaline teleosts in fresh water by promoting ion retention. In the euryhaline Mozambique tilapia (Oreochromis mossambicus), pituitary PRL cells release two PRL isoforms, PRL188 and PRL177, in response to a fall in extracellular osmolality. Both PRLs function via two PRL receptors (PRLRs) denoted PRLR1 and PRLR2. We conducted a comparative study using the Nile tilapia (O. niloticus), a close relative of Mozambique tilapia that is less tolerant to increases in environmental salinity, to investigate the regulation of PRLs and PRLRs upon acute hyperosmotic challenges in vivo and in vitro. We hypothesized that differences in the regulation of PRLs and PRLRs underlie the variation in salinity tolerance of tilapias within the genus Oreochromis. When transferred from fresh water to brackish water (20‰), Nile tilapia increased plasma osmolality and decreased circulating PRLs, especially PRL177, to a greater extent than Mozambique tilapia. In dispersed PRL cell incubations, the release of both PRLs was less sensitive to variations in medium osmolality in Nile tilapia than in Mozambique tilapia. By contrast, increases in pituitary and branchial prlr2 gene expression in response to a rise in extracellular osmolality were more pronounced in Nile tilapia relative to its congener, both in vitro and in vivo. Together, these results support the conclusion that inter-specific differences in salinity tolerance between the two tilapia congeners are tied, at least in part, to the distinct responses of both PRLs and their receptors to osmotic stimuli.

Cortisol is the final product of the hypothalamic-pituitary-interrenal (HPI) axis and acts as a gluco- and mineralo-corticoid in fish. Long-term elevations of cortisol have been linked to reduced growth in fishes, but the mechanism(s) and relative sensitivities of species are still unclear. We carried out experiments to examine the relative effects of cortisol on growth and gill NKA activity in two salmonids: Atlantic salmon (Salmo salar) and brook trout (Salvelinus fontinalis). Treatment with intraperitoneal cortisol implants for 30 days resulted in reduced growth in both species, but with greater sensitivity to cortisol in brook trout. Gill NKA activity was strongly upregulated by cortisol in Atlantic salmon, and weakly upregulated in brook trout but with no statistically significant effect. Cortisol treatment resulted in reduced plasma levels of insulin-like growth factor I and increased plasma growth hormone levels in Atlantic salmon. Our results demonstrate that there are species differences in the sensitivity of growth and osmoregulation to cortisol, even among species in the same family (Salmonidae).

Anadromous sea lamprey (Petromyzon marinus) larvae undergo a months-long true metamorphosis during which they develop seawater (SW) tolerance prior to downstream migration and SW entry. We have previously shown that intestinal Na+/K+-ATPase (NKA) activity increases during metamorphosis and is critical to the osmoregulatory function of the intestine in SW. The present study investigated the role of 11-deoxycortisol (S) in controlling NKA in the anterior (AI) and posterior (PI) intestine during sea lamprey metamorphosis. In a tissue profile, nka mRNA and protein were most abundant in the gill, kidney, and AI. During metamorphosis, AI nka mRNA increased 10-fold, whereas PI nka mRNA did not change. Specific corticosteroid receptors were found in the AI, which had a higher binding affinity for S compared to 11-deoxycorticosterone (DOC). In vivo administration of S in mid-metamorphic lamprey upregulated NKA activity 3-fold in the AI and PI, whereas administration of DOC did not affect intestinal NKA activity. During a 24 h SW challenge test, dehydration of white muscle moisture was rescued by prior treatment with S, which was associated with increased intestinal nka mRNA and NKA activity. These results indicate that intestinal osmoregulation in sea lamprey is a target for control by S during metamorphosis and the development of SW tolerance.

Gonadotropin-releasing hormone (GnRH) is considered a key player in reproduction. The caudal neurosecretory system (CNSS) is a unique neurosecretory structure of fish that may be involved in osmoregulation, nutrition, reproduction, and stress-related responses. However, a direct effect of GnRH on Dahlgren cells remains underexplored. Here, we examined the electrophysiological response of Dahlgren cell population of the CNSS to GnRH analog LHRH-A2 and the transcription of related key genes of CNSS. We found that GnRH increased overall firing frequency and may be changed the firing pattern from silent to burst or phasic firing in a subpopulation of Dahlgren cells. The effect of GnRH on a subpopulation of Dahlgren cells firing activity was blocked by the GnRH receptor (GnRH-R) antagonist cetrorelix. A positive correlation was observed between the UII and GnRH-R mRNA levels in CNSS or gonadosomatic index (GSI) during the breeding season. These findings are the first demonstration of the ability of GnRH acts as a modulator within the CNSS and add to our understanding of the physiological role of the CNSS in reproduction and seasonal adaptation.

Water deprivation of the Spinifex hopping mouse, Notomys alexis, induced a biphasic pattern of food intake with an initial hypophagia that was followed by an increased, and then sustained food intake. The mice lost approximately 20% of their body mass and there was a loss of white adipose tissue. Stomach ghrelin mRNA was significantly higher at day 2 of water deprivation but then returned to the same levels as water-replete (day 0) mice for the duration of the experiment. Plasma ghrelin was unaffected by water deprivation except at day 10 where it was significantly increased. Plasma leptin levels decreased at day 2 and day 5 of water deprivation, and then increased significantly by the end of the water deprivation period. Water deprivation caused a significant decrease in skeletal muscle leptin mRNA expression at days 2 and 5, but then it returned to day 0 levels by day 29. In the hypothalamus, water deprivation caused a significant up-regulation in both ghrelin and neuropeptide Y mRNA expression, respectively. In contrast, hypothalamic GHSR1a mRNA expression was significantly down-regulated. A significant increase in LepRb mRNA expression was observed at days 17 and 29 of water deprivation. This study demonstrated that the sustained food intake in N. alexis during water deprivation was uncoupled from peripheral appetite-regulating signals, and that the hypothalamus appears to play an important role in regulating food intake; this may contribute to the maintenance of fluid balance in the absence of free water.

International audience; Arginine vasotocin (AVT) is known to play an important role in the regulation of social behavior in a number of vertebrate species. Nevertheless, the relationship between AVT and intraspecific interactions appears complex and in some cases contradictory. Moreover, AVT influences other behaviors, which are not primarily social including exploratory behavior, locomotion and thermoregulation. Some of these behavioral effects may be side-effects from a general influence of AVT on physiology. Indeed AVT can regulate metabolism and osmoregulation. Because most studies have been conducted using mammals and birds, its role in modulating behavior in other vertebrate groups is largely unknown. In this study we examined the effect of AVT on the social behavior of male common lizards, Zootoca vivipara. Moreover, considering the variety of pathways AVT could be involved in, we investigated its consequences on thermoregulatory behavior and physiological performance. In mid June 2010, 74 males were captured from field sites (Mont-Lozere, South-eastern France) and kept in the laboratory for three weeks to obtain behavioral (reaction to conspecific odors, thermoregulation) and physiological (endurance, testosterone level) measurements. We demonstrated that injection of AVT reduced testosterone level and affected social behavior in different ways depending on the size of an individual. Specifically, small males injected with AVT were less attracted by conspecific odors than small control males, and no effect was detected in large males. Moreover, AVT promoted thermoregulatory behavior and enhanced endurance. These results are concordant with previous results obtained in this species in studies on stress suggesting that AVT may act through its influence on corticosterone secretion.

In fishes, prolactin (Prl) signaling underlies the homeostatic regulation of hydromineral balance by controlling essential solute and water transporting functions performed by the gill, gastrointestinal tract, kidney, urinary bladder, and integument. Comparative studies spanning over 60 years have firmly established that Prl promotes physiological activities that enable euryhaline and stenohaline teleosts to reside in freshwater environments; nonetheless, the specific molecular and cellular targets of Prl in ion- and water-transporting tissues are still being resolved. In this short review, we discuss how particular targets of Prl (e.g., ion cotransporters, tight-junction proteins, and ion pumps) confer adaptive functions to the esophagus and intestine. Additionally, in some instances, Prl promotes histological and functional transformations within esophageal and intestinal epithelia by regulating cell proliferation. Collectively, the demonstrated actions of Prl in the gastrointestinal tract of teleosts indicate that Prl operates to promote phenotypes supportive of freshwater acclimation and to inhibit phenotypes associated with seawater acclimation. We conclude our review by underscoring that future investigations are warranted to determine how growth hormone/Prl-family signaling evolved in basal fishes to support the gastrointestinal processes underlying hydromineral balance.

International audience; Plasma corticosterone (CORT) concentrations fluctuate in response to homeostatic demands. CORT is widely recognized as an important hormone related to energy balance. However, far less attention has been given to the potential role of CORT in regulating salt and water balance or responding to osmotic imbalances. We examined the effects of reproductive and hydric states on CORT levels in breeding Children’s pythons (Antaresia childreni), a species with substantial energetic and hydric costs associated with egg development. Using a 2 × 2 experimental design, we examined how reproduction and water deprivation, both separately and combined, impact CORT levels and how these changes correlate with hydration (plasma osmolality) and energy levels (blood glucose). We found that reproduction leads to increased CORT levels, as does dehydration induced by water deprivation. The combined impact of reproduction and water deprivation led to the largest increases in CORT levels. Additionally, we found significant positive relationships among CORT levels, plasma osmolality, and blood glucose. Our results provide evidence that both reproductive activity and increased plasma osmolality can lead to increased plasma CORT in an ectotherm, which could be explained by either CORT having a role as a mineralocorticoid or CORT being elevated as part of a stress response to resource imbalances.

International audience; Ticks can survive in harsh and fluctuating vegetated environments for long durations between blood feedings with highly developed osmoregulatory mechanisms. Like the unique life history of hematophagous ticks, osmoregulatory organs and their regulatory mechanisms are significantly different from those in the closely related insect taxa. Over the last ten years, research has uncovered several neuropeptidergic innervations of the primary osmoregulatory organ, the salivary glands: myoinhibitory peptide (MIP), SIFamide, and elevenin. These neuropeptides are thought to be modulators of dopamine's autocrine or paracrine actions controlling the salivary glands, including the activation of fluid transport into the lumen of salivary acini and the pumping and gating action of salivary acini for expelling fluids out into salivary ducts. These actions are through two different dopamine receptors, D1 receptor and invertebrate D1-like dopamine receptor, respectively. Interestingly, MIP and SIFamide are also involved in the control of another important excretory/osmoregulatory organ, the hindgut, where SIFamide is myostimulatory, with MIP having antagonistic effects. FGLamide related allatostatin is also found to have axonal projections located on the surface of the rectum. Investigations of the osmoregulatory mechanisms of these critical vector species will potentially lead to the development of a measure to control tick species.

The present study determined the effect of osmotic stress on the insulin-like receptor binding characteristics and on glucose metabolism in the anterior (AG) and posterior (PG) gills of the crab Neohelice granulata . Bovine insulin increased the capacity of the PG cell membrane to phosphorylate exogenous substrate poly (Glu:Tyr 4:1) and the glucose uptake in the control crab group. The crabs were submitted to three periods of hyperosmotic (HR) and hyposmotic (HO) stress, for 24, 72 and 144 h, to investigate the insulin-like receptor phosphorylation capacity of gills. Acclimation to HO for 24 h or HR for 144 h of stress inhibited the effects of insulin in the PG, decreasing the capacity of insulin to phosphorylate exogenous substrate poly (Glu:Tyr 4:1) and decreasing the glucose uptake. Hyperosmotic stress for the same period of 144 h significantly affected 125 I-insulin binding in the AG and PG. However, HO stress for 24 h significantly reduced 125 I-insulin-specific uptake only in the PG. Therefore, osmotic stress induces alterations in the gill insulin-like receptors that decrease insulin binding in the PG. These findings indicate that osmotic stress induced a pattern of insulin resistance in the PG. The free-glucose concentration in the PG decreased during acclimation to 144 h of HR stress and 24 h of HO stress. This decrease in the cell free-glucose concentration was not accompanied by a significant change in hemolymph glucose levels. In AG from the control group, neither the capacity of bovine insulin to phosphorylate exogenous substrate poly (Glu:Tyr 4:1) nor the glucose uptake changed; however, genistein decreased tyrosine-kinase activity, confirming that this receptor belongs to the tyrosine-kinase family. Acclimation to HO (24 h) or HR (144 h) stress decreased tyrosine-kinase activity in the AG. This study provided new information on the mechanisms involved in the osmoregulation process in crustaceans, demonstrating for the first time in an estuarine crab that osmotic challenge inhibited insulin-like signaling and the effect of insulin on glucose uptake in the PG.

Crustacean hyperglycemic hormones (CHHs) are multifunctional neuropeptides ubiquitous in crustaceans. In Litopenaeus vannamei, CHH-B2 is a CHH eyestalk isoform whose expression has been shown to vary with enviromental conditions, suggesting its relevance for ecophysiological performance of shrimp, controlling processes related to metabolism and osmo-ionic regulation. To study the involvement of CHH-B2 in these processes, we cloned and expressed a recombinant version with a free C-terminal glycine (rCHH-B2-Gly) in the methylotrophic yeast Pichia pastoris. The rCHH-B2-Gly peptide secreted to the culture medium was purified by RP-HPLC and used for in vivo glucose, triglyceride, and osmoregulation dose-response analyses with juvenile shrimp. The peptide was also amidated at the C-terminus using an α-amidating enzyme to produce rCHH-B2-amide. The shrimp showed a dose-dependent effect of rCHH-B2-Gly to hemolymph glucose and triglyceride levels, inducing maximal increases by injecting 500 and 1000pmol of hormone, respectively. Additionally, 10pmol of hormone was sufficient to reduce the hypo-osmoregulatory capacity of shrimp at 35‰. These findings suggest that CHH-B2 has regulatory roles in carbohydrate and lipid metabolism, and a potential involvement in osmoregulation of L. vannamei. Injection of 100pmol of rCHH-B2-amide increased glucose and triglyceride levels by 15 and 28%, respectively in comparison with rCHH-B2-Gly, suggesting an important role for the C-terminal amidation. Additionally, an in silico structural analysis done with the CHH-B1 and rCHH-B2-Gly peptides suggests that the C-terminal region may be relevant for the activity of the L. vannamei isoforms and explain the functional divergence from other crustacean CHH/CHH-like peptides.

It has long been held that cortisol, a glucocorticoid in many vertebrates, performs glucocorticoid and mineralocorticoid actions in the teleost fish since it lacks aldosterone. However, in addition to the counterparts of tetrapod mineralocorticoid receptors (MRs), 11-deoxycorticosterone (DOC) has been recently identified as a specific endogenous ligand for the MRs in teleosts. Here, we point out the minor role of mineralocorticoid signaling (i.e., DOC-MR) in the osmoregulation compared with those of glucocorticoid signaling (i.e., cortisol-glucocorticoid receptor [GR]), and review the current findings on the physiological roles of the DOC-MR in teleosts. Cortisol promotes both freshwater and seawater adaptation via the GRs in the osmoregulatory organs such as gills and gastrointestinal tracts, but the expressions of MR mRNA are abundant in the brains especially in the key components of the stress axis and cerebellums. Together with the behavioral effects of intracerebroventricular injection with DOC, the MR is suggested to play an important role in the brain dependent behaviors. Since the abundant expression of central MRs has been reported also in higher vertebrates and the MR is thought to be ancestral to the GR, the role of MR in fish might reflect the principal and original function of corticosteroid signaling. Functional evolution of corticosteroid systems is summarized and areas in need of research like our on-going experiments with MR-knockout medaka are outlined.

Adrenaline and cortisol, the major stress hormones, are known for its direct control on stress response in fish. Likewise, as an important stress modifier hormone, thyroid hormone has also been implicated in stress response of fish. We tested whether the hypothesis on the phenomenon of inter-hormonal interference, a process that explains the hormonal interactions, operates in fish particularly between adrenaline, cortisol and thyroid hormones. To achieve this goal, indices of acid-base, osmotic and metabolic regulations were quantified after adrenaline challenge in propranolol pre-treated air-breathing fish (Anabas testudineus). Short-term adrenaline (10 ng g(-1)) injection for 30 min produced a rise in plasma cortisol without affecting plasma T(3) and T(4). On the contrary, blocking of adrenaline action with a non-selective blocker, propranolol (25 ng g(-1)) for 90 min reduced plasma cortisol along with plasma T(4) and that indicate a possible interference of these hormones in the absence of adrenaline challenge. Similarly, a reduction in plasma T(3) was found after adrenaline challenge in propranolol pre-treated fish and that suggests a functional synergistic interference of adrenaline with T(3). Adrenaline challenge in these fish, however, failed to abolish this propranolol effect. The remarkable systemic hypercapnia and acidosis by propranolol pre-treatment were reversed by adrenaline challenge, pointing to a direct action of adrenaline on acid-base indices probably by a mechanism which may not require β-adrenergic receptor systems. Interestingly, the prominent adrenaline-induced hyperglycemia, hyperlactemia and hyperuremea were not altered by propranolol treatment. Similarly, adrenaline challenge promoted and propranolol reduced the osmotic competencies of the gills, kidneys and liver of this fish as evident in the sodium and proton pump activities. The modified physiologic actions of adrenaline and its modified interaction with THs and cortisol in blocked fish indicate an interaction of adrenaline with cortisol and THs. Our physiologic evidences thus support the hypothesis of the phenomenon of inter-hormonal interference.

Gene expression for growth hormone (GH)/prolactin (PRL)/somatolactin (SL) family hormones in the pituitaries of homing chum salmon were examined, because gene expression for these hormones during ocean-migrating phases remains unclear. Fish were collected in the winter Gulf of Alaska, the summer Bering Sea and along homing pathway in the Ishikari River-Ishikari Bay water system in Hokkaido, Japan in autumn. The oceanic fish included maturing adults, which had developing gonads and left the Bering Sea for the natal river by the end of summer. The absolute amounts of GH, PRL and SL mRNAs in the pituitaries of the maturing adults in the summer Bering Sea were 5- to 20-fold those in the winter Gulf of Alaska. The amount of GH mRNA in the homing adults at the coastal seawater (SW) areas was smaller than that in the Bering fish, while the amount of PRL mRNA remained at the higher level until fish arrived at the Ishikari River. The gill Na(+),K(+)-ATPase activity in the coastal SW fish and the plasma Na(+) levels in the brackish water fish at the estuary were lowered to the levels that were comparable to those in the fresh water (FW) fish. In conclusion, gene expression for GH, PRL and SL was elevated in the pituitaries of chum salmon before initiation of homing behavior from the summer Bering Sea. Gene expression for GH is thereafter lowered coincidently with malfunction of SW adaptability in the breeding season, while gene expression for PRL is maintained high until forthcoming FW adaptation.

In the present investigation, in vitro effects of vasotocin (VT) on oocyte (follicular) hydration during germinal vesicle breakdown (GVBD) and ovulation were demonstrated in hCG-primed and non-primed catfish. The data were compared with that of groups incubated with isotocin, and hCG alone or in combination with VT. The priming with hCG resulted in significant increases on percentage GVBD and ovulation, and stimulated follicular hydration, as judged by the increase in diameter, volume, water content, osmolality and Ca(2+) concentration. However, Na(+), K(+) ATPase activity, and concentrations of Na(+), K(+) and Mg(2+) did not alter significantly. The incubations with hCG or VT stimulated all the above parameters. In the non-primed fish, the response of hCG was significantly higher on follicular diameter, volume and osmolality, and that of VT on ovulation. In the primed fish, the VT response was significantly higher on GVBD, ovulation, Na(+), K(+) ATPase activity and divalent cation concentrations. The co-incubation with both hCG and VT produced maximal increases in all the parameters with significantly higher effects in the primed fish. The effects of IT on various parameters were relatively low compared to hCG or VT effects. The results indicate that VT may play an important role in oocyte (follicular) hydration, which is consistent with its role in osmoregulation of fish.

The removal of the eyestalk (s) induces molting and reproduction promoted the presence of regulatory substances in the eyestalk (ES), particularly medulla terminalis X-organ and the sinus gland (MTXO-SG). The PCR-based cloning strategies have allowed for isolating a great number of cDNAs sequences of crustacean hyperglycemic hormone (CHH) neuropeptides family from the eyestalk and non-eyestalk tissues, e.g., pericardial organs and fore- and hindguts. However, the translated corresponding neuropeptides in these tissues, their circulating concentrations, the mode of actions, and specific physiological functions have not been well described. The profiles of CHH neuropeptides present in the MTXO-SG may differ among decapod crustacean species, but they can be largely divided into two sub-groups on the basis of structural homology: (1) CHH and (2) molt-inhibiting hormone (MIH)/mandibular organ-inhibiting hormone (MOIH)/vitellogenesis/gonad-inhibiting hormone (V/GIH). CHH typically elevating the level of circulating glucose from animals under stressful conditions (hyper- and hypothermia, hypoxia, and low salinity) has multiple target tissues and functions such as ecdysteroidogenesis, osmoregulation, and vitellogenesis. Recently, MIH, known for exclusively suppressing ecdysteroidogenesis in Y-organs, is also reported to have an additional role in vitellogenesis of adult female crustacean species, suggesting that some CHH neuropeptides may acquire an extra regulatory role in reproduction at adult stage. This paper reviews the regulatory roles of CHH and MIH at the levels of specific functions, temporal and spatial expression, titers, their binding sites on the target tissues, and second messengers from two crab species: the blue crab, Callinectes sapidus, and the European green crab, Carcinus maenas. It further discusses the diverse regulatory roles of these neuropeptides and the functional plasticity of these neuropeptides in regard to life stage and species-specific physiology.

The responses of Mozambique and Nile tilapia acclimated to fresh water (FW) and brackish water (BW; 17 per thousand) were compared following acute salinity challenges. In both species, plasma osmolality increased to above 450 mOsm by 2h after transfer from FW to seawater (SW); these increases in osmolality were accompanied by unexpected increases in plasma prolactin (PRL). Likewise, PRL receptor gene expression in the gill also increased in both species. In Nile tilapia, hyperosmotic transfers (FW to BW and SW) resulted in increased plasma growth hormone (GH) and in branchial GH receptor gene expression, responses that were absent in Mozambique tilapia. Branchial gene expression of osmotic stress transcription factor 1 (OSTF1) increased in both species following transfer from FW to SW, whereas transfer from BW to SW induced OSTF1 expression only in the Nile tilapia. Branchial expression of Na(+)/Cl(-) cotransporter was higher in FW in both species than in BW. Branchial gene expression of Na(+)/K(+)/2Cl(-) cotransporter (NKCC) increased after transfer from BW to SW in Mozambique tilapia, whereas expression was reduced in the Nile tilapia following the same transfer. The difference in the SW adaptability of these species may be related to a limited capacity of Nile tilapia to up-regulate NKCC gene expression, which is likely to be an essential component in the recruitment of SW-type chloride cells. The differential responses of GH and OSTF1 may also be associated with the disparate SW adaptability of these two tilapiine species.

The natriuretic peptide (NP) family is a seemingly ubiquitous sodium and volume reducing endocrine system of predominantly cardiac origin. Members of the NP system include ANP, BNP, CNP, VNP, their guanylate cyclase (GC)-linked receptors (NPR-A and NPR-B), and clearance receptor (NPR-C). Through the activation of their membrane-bound GC receptors, these small peptides modulate cellular functions that affect both salt and water balance. The elucidation of piscine NP sequences, structure, and functions has steadily advanced over the past 15 years spearheaded by research from Dr. Yoshio Takei's laboratory. The development of these homologous NPs has led to extensive research into both the evolutionary and physiological significance of NPs in fishes. One outcome has been the development of two seemingly disparate hypotheses of NP function; a role in salt excretion, the osmoregulatory hypothesis, versus a role in protecting the heart, the cardioprotective hypotheses. In the osmoregulatory hypothesis NPs are released in response to elevated ambient salinity and inhibit drinking and intestinal uptake of salt, thereby effectively reducing plasma sodium levels. In contrast, the cardioprotective theory depicts NPs acting to prevent debilitating cardiodilation from an excess of either venous or arterial pressure through vasodilation and a reduction of blood volume. These seemingly distinct hypotheses may be elements of a more general regulatory system and certainly require further investigation. Undoubtedly their resolution will not only give us a better perspective of the evolutionary basis of the NP system but will provide us with a greater appreciation of salt and water homeostasis in vertebrates.

It has long been held that cortisol, acting through a single receptor, carries out both glucocorticoid and mineralocorticoid actions in teleost fish. The recent finding that fish express a gene with high sequence similarity to the mammalian mineralocorticoid receptor (MR) suggests the possibility that a hormone other than cortisol carries out some mineralocorticoid functions in fish. To test for this possibility, we examined the effect of in vivo cortisol, 11-deoxycorticosterone (DOC) and aldosterone on salinity tolerance, gill Na(+),K(+)-ATPase (NKA) activity and mRNA levels of NKA alpha 1a and alpha 1b in Atlantic salmon. Cortisol treatment for 6-14 days resulted in increased, physiological levels of cortisol, increased gill NKA activity and improved salinity tolerance (lower plasma chloride after a 24h seawater challenge), whereas DOC and aldosterone had no effect on either NKA activity or salinity tolerance. NKA alpha 1a and alpha 1b mRNA levels, which increase in response to fresh water and seawater acclimation, respectively, were both upregulated by cortisol, whereas DOC and aldosterone were without effect. Cortisol, DOC and aldosterone had no effect on gill glucocorticoid receptor GR1, GR2 and MR mRNA levels, although there was some indication of possible upregulation of GR1 by cortisol (p=0.07). The putative GR blocker RU486 inhibited cortisol-induced increases in salinity tolerance, NKA activity and NKA alpha 1a and alpha 1b transcription, whereas the putative MR blocker spironolactone had no effect. The results provide support that cortisol, and not DOC or aldosterone, is involved in regulating the mineralocorticoid functions of ion uptake and salt secretion in teleost fish.

Pacific salmon migrate from ocean through the natal river for spawning. Information on expression of genes encoding osmoregulatory hormones and migratory behavior is important for understanding of molecular events that underlie osmoregulation of homing salmon. In the present article, regulation of gene expression for osmoregulatory hormones in pre-spawning salmon was briefly reviewed with special reference to neurohypophysial hormone, vasotocin (VT), and pituitary hormones, growth hormone (GH) and prolactin (PRL). Thereafter, we introduced recent data on migratory behavior from SW to FW environment. In pre-spawning chum salmon, the hypothalamic VT mRNA levels increased in the males, while decreased in the females with loss of salinity tolerance when they were kept in SW. The amounts of GH mRNA in the pituitary decreased during ocean migration prior to entrance into FW. Hypo-osmotic stimulation by SW-to-FW transfer did not significantly affect the amount of PRL mRNA, but it was elevated in both SW and FW environments along with progress in final maturation. Behaviorally, homing chum salmon continued vertical movement between SW and FW layers in the mouth of the natal river for about 12h prior to upstream migration. Pre-spawning chum salmon in an aquarium, which allowed fish free access to SW and FW, showed that individuals with the lower plasma testosterone (T) and higher estradiol-17beta (E2) levels spent longer time in FW when compared with the SW fish. Taken together, neuroendocrine mechanisms that underlie salt and water homeostasis and migratory behavior from SW to FW may be under the control of the hypothalamus-pituitary-gonadal axis in pre-spawning salmon.

The life cycle of the European eel (Anguilla anguilla) includes two long migratory periods, when the newly hatched leptocephali larvae drift on ocean currents from the Sargasso Sea to the shores of Western Europe and then again up to 30 years later when adult eels swim back to their place of birth for reproductive purposes. Prior to the migration from fresh water (FW) to sea water (SW) adult yellow eels undergo various anatomical and physiological adaptations (silvering) which promote sexual development and aid the transition to increased environmental salinities. The aim of this study was to identify and characterise changes in gene expression within the major osmoregulatory tissues of the eel which enable these fish to make the physiological adaptations required for transfer to SW environments. In particular, changes in the expression of the FW-adapting hormone prolactin were correlated with differential expression of known osmoregulatory important genes within the gill, intestine and kidney following the acclimation of eels to SW. Various tissues were sampled from individual fish at selected intervals over a 5-month period following FW/SW transfer and RNA was isolated. Suppressive subtractive hybridization (SSH) was used for enrichment of differentially expressed genes. Microarrays comprising 6144 cDNAs spotted in triplicate, from brain, gill, intestine and kidney libraries (1536 randomly selected clones per tissue library), were hybridized with appropriate targets and analysed. Microarray results were validated using known genes implicated in osmoregulation, such as prolactin, growth hormone, Na, K-ATPase and some unknown genes, the role of which in osmoregulation needs to be elucidated.

Stress responses to increased temperature in black porgy reared in freshwater (FBP) and seawater (SBP) were examined via endocrinological and blood physiological methods. A rise in temperature increased plasma cortisol levels, which were significantly higher in FBP compared to SBP. The stimulated expression of phosphoenolpyruvate carboxykinase (PEPCK) mRNA in liver might result from the high cortisol level, and this explains the observed higher plasma glucose levels in FBP versus SBP. Full-length cDNA sequence for PEPCK was determined by 3' and 5' RACE procedures. PEPCK cDNA clone was found to contain 2563 nucleotides including an open reading frame that encodes 624 amino acids. While aspartate aminotransferase (AST) and alanine aminotransferase (ALT) of FBP increased with temperature, there was no change in SBP. In FBP, T(3) were 2.3+/-0.3 ng/ml at 20 degrees C and significantly decreased to 1.0+/-0.3 ng/ml at 30 degrees C. On the other hand, in SBP, it were 3.1+/-0.5 ng/ml at 20 degrees C but significantly increased to 5.2+/-0.4 ng/ml at 30 degrees C. When comparing osmolality at the temperature of 30 degrees C and of 20 degrees C, the difference was found to be greater for FBP than SBP. Accordingly, the results suggest that FBP suffers greater stress than SBP with increased temperature, and provide stress responses and osmoregulatory abilities against stressors in black porgy that could differ depending on salinities.

Larval-stage summer flounder (Paralichthys dentatus) were immersed in the corticosteroid-receptor blocker RU486 to test the effects of cortisol deficiency on salinity tolerance. Premetamorphic larvae held at 10 (near isosmotic) or 30 (hyperosmotic) parts per thousand ( per thousand) seawater survived well over 5d in 0, 0.012, or 0.12 microM RU486. However, at concentrations of 1.2 or 3.6 microM RU486, mortality was significantly greater for larvae in 30 per thousand compared to larvae in 10 per thousand. In a separate experiment, the ability of RU486 to inhibit tolerance to hyperosmotic medium (30 per thousand) was confirmed; immersion at 1.2 microM RU486 induced mortality of larvae in the metamorphic climax stage held at 30 per thousand, but not 0 or 10 per thousand. Mortality due to RU486 in pre- or prometamorphic stage larvae was prevented by concurrent immersion in cortisol at concentrations approximately 10-200 times greater than RU486, indicating that the action of RU486 was specific to antagonism of cortisol. The efficacy of 1.2 microM RU486 in reducing survival in 30 per thousand was found to be stage-dependent and exhibited the following hierarchy for fastest time to 50% mortality: prometamorphosismetamorphic climaxpremetamorphosis. In a 5-d pretreatment of pre- or prometamorphic larvae by immersion in 20 microM cortisol and/or 0.12 microM RU486 at 30 per thousand, only RU486 had a limited effect on decreasing survival when larvae were challenged with abrupt exposure to 50 per thousand. In total, the results evidence for the first time a necessary role for cortisol in seawater tolerance of a larval marine teleost.

Contains fulltext : 36596.pdf ( ) (Open Access) The peripheral conversion of the prohormone 3,5,3',5'-tetraiodothyronine (T4) to the biologically active 3,5,3'-triiodothyronine (T3), via enzymatic deiodination by deiodinases, is an important pathway in thyroid hormone metabolism. The aim of this study was to test if thyroid hormones and cortisol, as well as the outer ring deiodination (ORD) metabolic pathway, are involved in the osmoregulatory response of Senegalese sole (Solea senegalensis, Kaup 1858). We measured osmoregulatory and endocrine parameters in immature juveniles S. senegalensis acclimated to seawater (SW, 38 per thousand) and that were transferred and allowed to acclimate to different salinities (5 per thousand, 15 per thousand, 38 per thousand and 55 per thousand) for 17 days. An adjustment and a chronic regulatory period were identified following acclimation. The adjustment period immediately follows the transfer, and is characterized by altered plasma osmolalities. During this period, plasma cortisol levels increased while plasma free T4 (fT4) levels decreased. Both hormones levels returned to normal values on day 3 post-transfer. In the adjustment period, renal and hepatic ORD activities had increased concomitantly with the decrease in plasma fT4 levels in fishes transferred to extreme salinities (5 per thousand and 55 per thousand). In the chronic regulatory period, where plasma osmolality returned to normal values, plasma cortisol had increased, whereas plasma fT4 levels decreased in animals that were transferred to salinities other than SW. No major changes were observed in branchial ORD activity throughout the experiment. The inverse relationship between plasma cortisol and fT4 suggests an interaction between these hormones during both osmoregulatory periods while ORD pathway can be important in the short-term adjustment period. 8 p.

Endocrine control of osmoregulation is essential for teleosts to adapt to various aquatic environments. Prolactin (PRL) is known as a fundamental endocrine factor for hyperosmoregulation in teleost fishes, acting on ionocytes in the gills to maintain ion concentrations of body fluid within narrow physiological ranges in freshwater conditions. Cortisol is also known as an osmoregulation-related steroid in teleosts; however, its precise function is still controversial. Here, we investigated more detailed effects of PRL and roles of cortisol on ionocytes of Mozambique tilapia (Oreochromis mossambicus) in freshwater, using an improved gill filament incubation system. This incubation system resulted in enhanced cell viability, as evaluated using the dead cell marker propidium iodide. PRL was shown to maintain the density of freshwater-type ionocytes in isolated gill filaments; this effect of PRL is not achieved by the activation of cell proliferation, but by the maintenance of existing ionocytes. Cortisol alone did not show any distinct effect on ionocyte density in isolated gill filaments. We also assessed effects of PRL and cortisol on relative mRNA levels of NCC2, NHE3, NKAa1a, and NKAa1b. PRL maintained relative NCC2 and NKAa1a mRNA abundance, and cortisol showed a stimulatory effect on relative NCC2 and NKAa1a mRNA levels in combination with PRL, though cortisol alone exerted no effect on these genes. An increase in NKAa1b mRNA abundance was detected in cortisol-treated groups. PRL treatment also maintained normal NCC2 localization at the apical membrane of the ionocytes. These results indicate that PRL maintains freshwater-type ionocytes, and that cortisol stimulates the function of ionocytes maintained by PRL.

Many Australian tetrapods inhabit desert environments characterised by low productivity, unpredictable rainfall, high temperatures and high incident solar radiation. Maintaining a homeostatic milieu interieur by osmoregulation and thermoregulation are two physiological challenges faced by tetrapods in deserts, and the endocrine system plays an important role in regulating these processes. There is a considerable body of work examining the osmoregulatory role of antidiuretic hormones for Australian amphibians, reptiles and mammals, with particular contributions concerning their role and function for wild, free-living animals in arid environments. The osmoregulatory role of the natriuretic peptide system has received some attention, while the role of adrenal corticosteroids has been more thoroughly investigated for reptiles and marsupials. The endocrinology of thermoregulation has not received similar attention. Reptiles are best-studied, with research examining the influence of arginine vasotocin and melatonin on body temperature, the role of prostaglandins in heart rate hysteresis and the effect of melanocyte-stimulating hormone on skin reflectivity. Australian mammals have been under-utilised in studies examining the regulation, development and evolution of endothermy, and there is little information concerning the endocrinology of thermoregulation for desert species. There is a paucity of data concerning the endocrinology of osmoregulation and thermoregulation for Australian desert birds. Studies of Australian desert fauna have made substantial contributions to endocrinology, but there is considerable scope for further research. A co-ordinated approach to examine arid-habitat adaptations of the endocrine system in an environmental and evolutionary context would be of particular value.

The osmoregulatory and metabolic role of testosterone (T) in the euryhaline teleost Sparus auratus was examined. Fish were implanted with a slow-release coconut oil implant alone (control) or containing T (2 or 5microgg(-1) body weight) and sampled 1, 3, and 7 days after implantation. Gill Na(+),K(+)-ATPase activity increased in fish treated with the lower dose of T after 7 days of treatment. Kidney Na(+),K(+)-ATPase activity enhanced at first day post-implantation in the group treated with the higher dose of T but the values diminished by day 3. Plasma levels of metabolites (glucose, lactate, triglyceride, and protein) increased after T treatment. This higher availability of plasma metabolites was reflected in several metabolic changes within different tissues of T-treated fish such as (i) increased glycogen levels and capacity for gluconeogenesis, ketogenesis, glucose exporting, and amino acid catabolism in the liver, (ii) enhanced lipogenic capacity in the gills, (iii) increased glycogen levels and capacity for oxidizing amino acids in the kidney, and (iv) enhanced levels of glycogen, aceotacetate, glucose and triglycerides, and higher capacity of phosphorylating glucose in the brain. These results provide evidence regarding an osmoregulatory and metabolic role for T in S. auratus that could be related to changes in both processes during sexual maturation.

Amphibians occupy a wide variety of ecological habitats, and their adaptation is made possible through the specialization of the epithelia of their osmoregulatory organs, such as the skin, kidney, and urinary bladder, which control the hydromineral and acid-base balance of their internal medium. Amphibians can change drastically plasma Na+, Cl-, and urea levels and excretion rates in response to environmental stimuli such as acute desiccation and changes in external salinity. Several hormones and the autonomic nervous system act to control osmoregulation. Several ion channels including an epithelial sodium channel (ENaC), a urea transporter (UT), and water channels (AQPs) are found in epithelial tissues of their osmoregulatory organs. This mini review examines the currents status of our knowledge about hormone receptors for arginine vasotocin, angiotensin II and aldosterone, and membrane ion channels and transporters, such as ENaC, UT, and AQPs in amphibians.

This study examined the effect of transfer to increased environmental salinity on the circulating levels of angiotensin II (ANG II), C-type natriuretic peptide (CNP), and arginine vasotocin (AVT) in the euryhaline elasmobranch, Carcharhinus leucas. Plasma levels of ANG II and CNP were significantly increased in C. leucas chronically acclimated to seawater (SW) in comparison to freshwater (FW) acclimated fish. There was no difference in plasma AVT levels. Acute transfer of FW fish to 75% SW induced an increase in plasma ANG II levels within 12 h, and subsequent transfer from 75 to 100% SW further increased plasma ANG II levels at both 24 and 72 h. No change in plasma CNP was observed during acute transfer to increased salinity. However, a significant increase in plasma AVT levels was observed following 96 h in 75% SW and 24 h in 100% SW. In chronically SW acclimated C. leucas plasma osmolality, sodium, chloride, and urea were all significantly higher than FW acclimated fish but there was no difference in haematocrit. Acute transfer of C. leucas to 75% SW induced a significant increase in plasma osmolality, sodium and urea concentrations within 96 h of transfer. Subsequent transfer from 75 to 100% SW induced a further increase in these variables within 24 h in addition to a significant increase in plasma chloride above control levels. Haematocrit did not differ between the experimental and control groups throughout the acute study. Circulating levels of ANG II were significantly correlated to plasma, sodium, chloride, and urea concentrations during acclimation to SW. Conversely, circulating levels of CNP and AVT did not correlate to plasma osmolytes, however, CNP was significantly correlated to haematocrit during acclimation to seawater.

Sex steroids are known to interfere with the parr-smolt transformation of anadromous salmonids, and environmental estrogens such as nonylphenol have recently been implicated in reduced returns of Atlantic salmon in the wild. To determine the endocrine pathways by which estrogenic compounds affect smolt development and seawater tolerance, groups of juvenile Atlantic salmon were injected with one of five doses (0.5, 2, 10, 40 or 150 μg g −1 ) of branched 4-nonylphenol (NP), 2 μg g −1 of 17β-estradiol (E 2 ), or vehicle, during the parr-smolt transformation in April, and the treatment was repeated 4, 8, and 11 days after the first injection. Plasma was obtained for biochemical analysis 7 and 14 days after initiation of treatment. After 14 days of treatment, additional fish from each treatment group were exposed to seawater for 24 h to assess salinity tolerance. The E 2 treatment and the highest NP dose resulted in lower salinity tolerance and decreased plasma insulin-like growth factor I (IGF-I) levels, along with elevated levels of plasma vitellogenin and total calcium. Plasma growth hormone levels were elevated at intermediate NP doses only, and not affected by E 2 . After 7 days, plasma thyroxine (T 4 ) levels decreased in a strong, dose-dependent manner in response to nonylphenol, but after 14 days, this suppressive effect of T 4 occurred at the highest NP dose only. Similarly, E 2 decreased plasma T 4 levels at 7, but not 14 days. Plasma 3,3′,5-triodo- l -thyronine was reduced by E 2 and the highest NP dose after 7 and 14 days of treatment. Plasma cortisol levels were not affected by any of the treatments. The results indicate that the parr-smolt transformation and salinity tolerance can be compromised by exposure to estrogenic compounds. Suppression of plasma IGF-I levels is a likely endocrine pathway for the effects of estrogenic compounds on hypo-osmoregulatory capacity, and the detrimental effects of E 2 and NP on thyroid hormone levels are also likely to compromise the normal parr-smolt transformation of Atlantic salmon.

To clarify the physiological roles of the renin-angiotensin-aldosterone system (RAAS) and arginine vasotocin (AVT) on body fluid regulation in amphibians, we measured plasma concentrations of aldosterone (ALDO), angiotensin II (ANG II), and AVT after various osmotic challenges in the marine toad, Bufo marinus (Bufonidae). Hematocrit value (Ht) as an indicator of plasma volume, plasma osmolality and concentrations of plasma components (Na(+), Cl(-), K(+), and urea) were also measured. The toads were maintained under various osmotic treatments for 7 days. In dehydrated toads, plasma concentrations of ALDO, ANG II, AVT, and all plasma components measured were increased. In toads maintained in 300 mosmol/kg H(2)O NaCl solution, plasma osmolality, Na(+), Cl(-), urea, and plasma AVT concentrations were significantly increased, and Ht and plasma concentrations of ALDO and ANG II were significantly decreased. In toads maintained in tap water, plasma osmolality, and concentrations of Na(+) and ALDO were significantly decreased. We also estimated total body water (TBW), plasma volume (PV) using Evans Blue dye and Ht in the toads under various osmotic treatments. In dehydrated toads, TBW and PV were significantly decreased and Ht was significantly increased in comparison with those of control. In toads maintained in 300 mosmol/kg H(2)O NaCl solution, TBW and PV were significantly increased and Ht was significantly decreased in comparison with those of control. There was a significant negative correlation between Ht and PV or TBW. These results show that dehydration, which induces hypovolemic and hyperosmotic conditions, stimulates increases of plasma ALDO, ANG II, and AVT concentrations, while hypervolemic treatment induces decreases of plasma ALDO and ANG II concentrations. There were significant correlations between plasma osmolality and AVT concentration, between Ht and concentrations of RAAS hormones, and between plasma concentrations of ALDO and ANG II. These results suggest that volumetric and osmometric systems regulated by RAAS hormones and AVT are present in B. marinus.

Sharks and rays utilize a unique strategy for adaptation to the hyperosmotic marine environment by maintaining their plasma slightly hyperosmotic to surrounding seawater (SW) through the accumulation of urea. Since neurohypophysial hormones (NHs) are plausible candidates for osmoregulatory effectors, the synthesis and release of NHs were investigated after transfer of fish to different environmental salinities. Molecular cloning revealed three NHs from the hypothalamus of a dogfish, Triakis scyllium: vasotocin (VT), asvatocin, and a novel oxytocin-family peptide, phasitocin ([Phe3, Asn4, Ile8]vasotocin). The VT precursor consists of a signal peptide, VT, a neurophysin and a copeptin moiety. In contrast, the asvatocin and phasitocin precursors are shorter due to the lack of a copeptin moiety as is the case in oxytocin and mesotocin precursors in tetrapods and lungfish, but different from teleost isotocin precursors that have the copeptin moiety. In the hypothalamus, VT mRNA levels significantly increased after transfer to concentrated (130%) SW for 2 days, while no change was observed in mRNA levels of asvatocin and phasitocin following transfer to either 130% or diluted (60%) SW. The increase in VT mRNA was reflected in the plasma level of peptide; plasma VT concentration measured by highly sensitive and specific radioimmunoassay increased according to elevated environmental salinities. These results suggest that VT is an osmoregulatory effector in dogfish, especially when the dogfish is exposed to a hyperosmotic environment.

Prolactin (PRL) plays a central role in freshwater (FW) adaptation in teleost fish. Evidence now suggests that growth hormone (GH) acts in the seawater (SW) adaptation in at least some euryhaline fish. Reflecting its important role in FW adaptation, plasma levels of PRL188 and PRL177 are higher in tilapia (Oreochromis mossambicus) adapted to FW than in those adapted to SW. A transient but significant increase in plasma GH was observed 6 h after transfer from FW to SW. Elevated plasma PRL levels were seen in association with reductions in plasma osmolality after blood withdrawal in FW fish whereas no significant change was seen in plasma GH levels. When pituitaries from FW tilapia were incubated for 7 days, secretion of both PRLs was significantly greater in hyposmotic medium than in hyperosmotic medium for the first 24 h. Secretion of GH from the same pituitary was relatively low during this period compared with PRL secretion. No consistent effect of medium osmolality on GH release was seen for the first day, but its cumulative release was increased significantly in hyperosmotic medium after 2 days and thereafter. On the other hand, ACTH release was extremely low compared with the secretion of PRLs and GH and there was no consistent effect of medium osmolality. These results indicate that PRL release from the tilapia pituitary is stimulated both in vivo and in vitro as extracellular osmolality is reduced, whereas the secretion of GH increases temporarily when osmolality is increased. ACTH seems to be relatively insensitive to the changes in environmental osmolality.

Plasma concentrations and stored levels of the neuroendocrine peptides arginine vasotocin (AVT) and urotensin II (UII) were measured in the euryhaline flounder (Platichthys flesus) following the acute hypo-osmotic challenge of direct seawater (SW) to fresh water (FW) transfer. Hormone measures, plasma osmolality, and ion concentrations and tissue water content were determined 1, 4, 8, 24, 72, and 144 h after transfer. Plasma AVT concentration fell initially following FW transfer but then returned toward pretransfer levels by day 6. Plasma UII concentration decreased while urophysial UII content was increased following hypo-osmotic challenge relative to SW time-matched controls, suggesting down regulation of the UII system during the initial stages after FW transfer. These changes in neuroendocrine activity were associated with a significant fall in plasma osmolality and major plasma ions. Positive correlations were observed between plasma AVT and osmolality and Cl- and Mg2+ concentrations, suggesting functional association of these plasma parameters with AVT action and/or control of AVT secretion. The initial response to hypotonic challenge involves reduced plasma AVT and UII levels consistent with the proposed role for these hormones, supporting flounder osmoregulation in hypertonic media.

It is thought that the elasmobranch corticosteroid hormone 1α-hydroxycorticosterone (1α-B) functions as both a glucocorticoid (GC) and mineralocorticoid (MC). Classical antinatriuretic MC activities would run counter to the osmoregulatory strategy of euryhaline elasmobranchs acclimating to fresh water (FW). Therefore we hypothesize that FW acclimation will be accompanied by a decrease in plasma corticosteroids in these animals. However, events that activate the "fight-or-flight" response could mask changes associated with acclimation to lower salinities. To better define the MC role of corticosteroids in elasmobranchs, we designed a transfer system that allows the acclimation of Atlantic stingrays (Dasyatis sabina) from seawater (SW) to FW over 12h while minimizing other extraneous stressors. Blood and interrenal glands were sampled from one group of stingrays 24h after FW transfer, while another group was sampled two weeks after FW transfer. Two other groups served as mock-transfer controls in that they were treated and sampled in the same way, but remained in SW for the entire period. Plasma corticosteroids, osmolality, chloride, and urea were significantly lower in FW-acclimated stingrays (compared to mock-transfer stingrays) 24h after FW transfer. This pattern remained after two weeks in FW, with the exception that plasma corticosteroids returned to pre-acclimation levels. There were no significant differences between experimental groups in interrenal levels of mRNAs encoding key steroidogenic proteins (steroidogenic acute regulatory protein and cholesterol side chain cleavage enzyme). Temporally decreased corticosteroid levels during FW acclimation are consistent with the unique strategy of euryhaline elasmobranchs, whereby lower plasma osmolality is maintained in FW vs. SW environments to reduce hydromineral gradients.

The native distribution of Mozambique tilapia, Oreochromis mossambicus, is characterized by estuarine areas subject to salinity variations between fresh water (FW) and seawater (SW) with tidal frequency. Osmoregulation in the face of changing environmental salinity is largely mediated through the neuroendocrine system and involves the activation of ion uptake and extrusion mechanisms in osmoregulatory tissues. We compared plasma osmolality, plasma prolactin (PRL), pituitary PRL mRNA, and mRNA of branchial ion pumps, transporters, channels, and PRL receptors in tilapia reared in FW, SW, brackish water (BW) and in tidally-changing salinity, which varied between FW (TF) and SW (TS) every 6h. Plasma PRL was higher in FW tilapia than in SW, BW, TF, and TS tilapia. Unlike tilapia reared in FW or SW, fish in salinities that varied tidally showed no correlation between plasma osmolality and PRL. In FW fish, gene expression of PRL receptor 1 (PRLR1), Na(+)/Cl(-) cotransporter (NCC), aquaporin 3 (AQP3) and two isoforms of Na(+)/K(+)-ATPase (NKA α1a and NKA α1b) was higher than that of SW, BW or tidally-changing salinity fish. Gene expression of the Na(+)/K(+)/2Cl(-) cotransporter (NKCC1a), and the cystic fibrosis transmembrane conductance regulator (CFTR) were higher in fish in SW, BW or a tidally-changing salinity than in FW fish. Immunocytochemistry revealed that ionocytes of fish in tidally-changing salinities resemble ionocytes of SW fish. This study indicated that tilapia reared in a tidally-changing salinity can compensate for large changes in external osmolality while maintaining osmoregulatory parameters within a narrow range closer to that observed in SW-acclimated fish.