Erratum: Direct Observation of Proton Emission in

This corrects the article DOI: 10.1103/PhysRevLett.123.082501.

Absence of Low-Energy Shape Coexistence in

The ^{80}Ge structure was investigated in a high-statistics ß-decay experiment of ^{80}Ga using the GRIFFIN spectrometer at TRIUMF-ISAC through γ, ß-e, e-γ, and γ-γ spectroscopy. No evidence was found for the recently reported 0_{2}^{+} 639-keV level suggested as evidence for low-energy shape coexistence in ^{80}Ge. Large-scale shell model calculations performed in ^{78,80,82}Ge place the 0_{2}^{+} level in ^{80}Ge at 2 MeV. The new experimental evidence combined with shell model predictions indicate that low-energy shape coexistence is not present in ^{80}Ge.

Direct Observation of Proton Emission in

The elusive ß^{-}p^{+} decay was observed in ^{11}Be by directly measuring the emitted protons and their energy distribution for the first time with the prototype Active Target Time Projection Chamber in an experiment performed at ISAC-TRIUMF. The measured ß^{-}p^{+} branching ratio is orders of magnitude larger than any previous theoretical model predicted. This can be explained by the presence of a narrow resonance in ^{11}B above the proton separation energy.

Effect of parental mate choice and semi-natural early rearing environment on the growth performance and seawater tolerance of Chinook salmon Oncorhynchus tshawytscha.

To assess whether parental mate choice and early rearing in a semi-natural spawning channel may benefit the culture of Chinook salmon Oncorhynchus tshawytscha, 90 day growth trials were conducted using hatchery O. tshawytscha (hatchery), mate choice O. tshawytscha (i.e. the offspring of parents allowed to choose their own mate) that spent 6 months in a spawning channel prior to hatchery rearing (channel) and mate choice O. tshawytscha transferred to the hatchery as fertilized eggs (transfer). During the growth trials, all O. tshawytscha stocks were reared separately or in either mixed channel and hatchery or transfer and hatchery groups for comparison of performance to traditional practices. After 60 days in fresh water, all O. tshawytscha were transferred to seawater for an additional 30 days. Reared separately, all stocks grew c. 4.5 fold over 90 days but specific growth rate (G) and food conversion efficiency were higher in fresh water than after seawater transfer on day 60. In contrast, hatchery O. tshawytscha from mixed hatchery and channel and hatchery and transfer growth trials had a larger mass and length gain than their counterparts on day 60, but reduced G in seawater. In general, plasma levels of growth hormone, insulin-like growth factor I and cortisol did not differ among any O. tshawytscha groups in either the separate or mixed growth trials. Despite some differences in gill Na(+),K(+)-ATPase activity, all O. tshawytscha had a high degree of seawater tolerance and experienced virtually no perturbation in plasma chloride following seawater transfer. Overall, all O. tshawytscha exhibited similar growth and seawater performance under traditional hatchery conditions and any benefit derived from either parental mate choice or semi-natural early rearing environment was only observed in the presence of mutual competition with hatchery O. tshawytscha.

Inhibition of glutamine synthetase during ammonia exposure in rainbow trout indicates a high reserve capacity to prevent brain ammonia toxicity.

Glutamine synthetase (GSase), the enzyme that catalyses the conversion of glutamate and ammonia to glutamine, is present at high levels in vertebrate brain tissue and is thought to protect the brain from elevated ammonia concentrations. We tested the hypothesis that high brain GSase activity is critical in preventing accumulation of brain ammonia and glutamate during ammonia loading in the ammonia-intolerant rainbow trout. Trout pre-injected with saline or the GSase inhibitor methionine sulfoximine (MSOX, 6 mg kg(-1)), were exposed to 0, 670 or 1000 micromol l(-1) NH(4)Cl in the water for 24 and 96 h. Brain ammonia levels were 3- to 6-fold higher in ammonia-exposed fish relative to control fish and MSOX treatment did not alter this. Brain GSase activity was unaffected by ammonia exposure, while MSOX inhibited GSase activity by approximately 75%. Brain glutamate levels were lower and glutamine levels were higher in fish exposed to ammonia relative to controls. While MSOX treatment had little impact on brain glutamate, glutamine levels were significantly reduced by 96 h. With ammonia treatment, significant changes in the concentration of multiple other brain amino acids occurred and these changes were mostly reversed or eliminated with MSOX. Overall the changes in amino acid levels suggest that multiple enzymatic pathways can supply glutamate for the production of glutamine via GSase during ammonia exposure and that alternative transaminase pathways can be recruited for ammonia detoxification. Plasma cortisol levels increased 7- to 15-fold at 24 h in response to ammonia and MSOX did not exacerbate this stress response. These findings indicate that rainbow trout possess a relatively large reserve capacity for ammonia detoxification and for preventing glutamate accumulation during hyperammonaemic conditions.

Evolution of ELNES spectra as a function of experimental settings for any uniaxial specimen: a fully relativistic study.

We perform calculations of the fully relativistic, corrected geometrical weighting of the pi* and sigma* transitions measured from the 1s core loss electron energy loss spectroscopy (EELS) spectrum in any uniaxial specimen. We present a complete calculation of the differential scattering cross-section (DSCS), taking into account the collection angle, the illumination angle and the tilt of the sample over the optical axis. Owing to high electron velocity in an EELS experiment, the relativistic correction has to be considered. We thus, present a relativistic, corrected DSCS by using the theory recently developed by Jouffrey et al. [Ultramicroscopy 102 (2004) 61] and P. Schattschneider et al. [Phys. Rev. B 72 (2005) 045142]. The relativistic correction is first performed in the natural coordinate system of the scattering event. We then point out a straightforward method to introduce this correction in the microscopic coordinate system, where all calculations have to be done to be experimentally useful. Using the fully corrected DSCS, we present an expression predicting the evolution of the R=pi*/(pi*+sigma*) ratio (related to the ratio of sp2 and sp3 bondings) as a function of experimental settings. We show how the R-evolution can be predicted, for any experimental setting, by the knowledge of one unique reference value. We verify on graphite specimens, the validity of the R-calculation by comparing theoretical predictions presented in this work with experimental data published elsewhere [Daniels et al., Ultramicroscopy 96 (2003) 523 and Menon et al., Ultramicroscopy 74 (1998) 83].

Nonreciprocal reconfigurable microwave optomechanical circuit.

Nonreciprocal microwave devices are ubiquitous in radar and radio communication and indispensable in the readout chains of superconducting quantum circuits. Since they commonly rely on ferrite materials requiring large magnetic fields that make them bulky and lossy, there has been significant interest in magnetic-field-free on-chip alternatives, such as those recently implemented using the Josephson nonlinearity. Here, we realize reconfigurable nonreciprocal transmission between two microwave modes using purely optomechanical interactions in a superconducting electromechanical circuit. The scheme relies on the interference in two mechanical modes that mediate coupling between the microwave cavities and requires no magnetic field. We analyse the isolation, transmission and the noise properties of this nonreciprocal circuit. Finally, we show how quantum-limited circulators can be realized with the same principle. All-optomechanically mediated nonreciprocity demonstrated here can also be extended to directional amplifiers, and it forms the basis towards realizing topological states of light and sound.Nonreciprocal optical devices traditionally rely on magnetic fields and magnetic-free approaches are rather recent. Here, Bernier et al. propose and demonstrate a purely optomechanical circulator with reconfigurable transmission without the need for direct coupling between input and output modes.

Mediation of humoral catecholamine secretion by the renin-angiotensin system in hypotensive rainbow trout (Oncorhynchus mykiss).

The individual contributions of, and potential interactions between, the renin-angiotensin system (RAS) and the humoral adrenergic stress response to blood pressure regulation were examined in rainbow trout. Intravenous injection of the smooth muscle relaxant, papaverine (10 mg/kg), elicited a transient decrease in dorsal aortic blood pressure (PDA) and systemic vascular resistance (RS), and significant increases in plasma angiotensin II (Ang II) and catecholamine concentrations. Blockade of alpha-adrenoceptors before papaverine treatment prevented PDA and RS recovery, had no effect on the increase in plasma catecholamines, and resulted in greater plasma Ang II concentrations. Administration of the angiotensin-converting enzyme inhibitor, lisinopril (10(-4) mol/kg), before papaverine treatment attenuated the increases in the plasma concentrations of Ang II, adrenaline, and noradrenaline by 90, 79, and 40%, respectively and also prevented PDA and RS recovery. By itself, lisinopril treatment caused a gradual and sustained decrease in PDA and RS, and reductions in basal plasma Ang II and adrenaline concentrations. Bolus injection of a catecholamine cocktail (4 nmol/kg noradrenaline plus 40 nmol/kg adrenaline) in the lisinopril+papaverine-treated trout, to supplement their circulating catecholamine concentrations and mimic those observed in fish treated only with papaverine, resulted in a temporary recovery in PDA and RS. These results indicate that the RAS and the acute humoral adrenergic response are both recruited during an acute hypotensive stress, and have important roles in the compensatory response to hypotension in rainbow trout. However, whereas the contribution of the RAS to PDA recovery is largely indirect and relies on an Ang II-mediated secretion of catecholamines, the contribution from the adrenergic system is direct and relies at least in part on plasma catecholamines.

Control of catecholamine and serotonin release from the chromaffin tissue of the Atlantic hagfish

An in situ saline-perfused systemic heart/posterior cardinal vein preparation of the Atlantic hagfish (Myxine glutinosa) was used to assess (1) the ability of the chromaffin tissue to release catecholamines in response to adrenocorticotropic hormone (ACTH; 7.5 i.u. kg-1), serotonin (250 nmol kg-1), carbachol (100 µmol kg-1), [Asn1-Val5]angiotensin II (Ang II; 100 nmol kg-1), histamine (0.3­300 µmol l-1) and a high-[K+] saline (60 mmol l-1), (2) whether serotonin is co-released with the catecholamines of the chromaffin tissues, and (3) the potential modulatory effects of NECA, an adenosine receptor agonist, and DPSPX, an adenosine receptor antagonist, on catecholamine release. Bolus injections of ACTH, serotonin or carbachol, or perfusion with high-[K+] saline, all elicited the release of both adrenaline and noradrenaline. Pre-treatment with the serotonergic receptor antagonist methysergide or the cholinergic receptor antagonist hexamethonium abolished the serotonin- and carbachol-mediated catecholamine releases, respectively. Neither receptor antagonist affected the ACTH-mediated catecholamine release. Bolus injections of Ang II or perfusion with a range of histamine concentrations, two potent secretagogues in other vertebrates, did not elicit catecholamine secretion in hagfish. While injections of Ang II or perfusion with the high-[K+] saline both elicited the release of serotonin, treatments with ACTH, carbachol or histamine did not. Hence, co-release of catecholamines and serotonin was elicited by non-specific cell membrane depolarization using K+, but not by the specific secretagogues assessed in this study. The adenosine receptor agonist NECA and antagonist DPSPX significantly modified the secretory responses elicited by ACTH, serotonin and carbachol. The results suggest that adenosine may inhibit catecholamine release induced by serotonin or carbachol, while stimulating ACTH-induced release. Although the contribution of the different secretagogues identified in this study has yet to be explored in vivo, our results suggest that the control of catecholamine and serotonin release from the aneural chromaffin tissue of the Atlantic hagfish can be achieved through hormonal and/or paracrine means.

Adenosine receptor blockade and hypoxia-tolerance in rainbow trout and Pacific hagfish. II. Effects on plasma catecholamines and erythrocytes

The purpose of this study was to examine the role of adenosine receptors (ARs) in (1) the regulation of catecholamine secretion and (2) the modulation of blood oxygen capacitance by catecholamines. To this end, we assessed the response of rainbow trout and Pacific hagfish treated with either an AR blocker, theophylline, or saline under hypoxic and normoxic conditions. Compared with the control hypoxic rainbow trout, AR blockade resulted in a smaller increase in haematocrit and haemoglobin (Hb) concentration of the blood, smaller red blood cell transmembrane pH differences and mean cellular [Hb] (MCHC), as well as a 16-fold higher plasma adrenaline concentration after only 10 min of acute hypoxic exposure. In hypoxic hagfish, AR blockade had no effect on the [Hb] of the blood, and there was no regulation of red blood cell pH or changes in MCHC. However, whereas plasma [adrenaline] did not change following exposure to a PwO2 of 1.33 kPa in the hypoxic sham group, the concentration increased 3.8-fold within 10 min in the theophylline-injected group. These results suggest that adenosine modulates the circulating level of catecholamines in both hypoxic rainbow trout and hypoxic Pacific hagfish.

Adenosine receptor blockade and hypoxia-tolerance in rainbow trout and Pacific hagfish. I. Effects on anaerobic metabolism

The physiological properties of adenosine may be essential in the control of energy metabolism for the survival of animals exposed to oxygen shortages. Accordingly, we tested the hypothesis that adenosine modulates metabolic regulation in rainbow trout and Pacific hagfish exposed to acute hypoxia. Treatment of hypoxic rainbow trout (PwO2=3.33 or 4.00 kPa) with the adenosine receptor (AR) blocker theophylline was associated with greater increases in plasma [lactate], more rapid and pronounced metabolic acidosis, higher tissue [lactate], and lower heart creatine charge and glycogen content than in the hypoxic controls. The recruitment of anaerobic metabolism in hypoxic trout treated with enprofylline, an AR blocker with very weak affinity, was intermediate to that of the hypoxic theophylline-injected and control groups. In hagfish, plasma [lactate] increased following exposure to a PwO2 of 1.33 kPa but did not increase following exposure to 3.33 kPa and, like plasma acidosis, it was greatest in the animals treated with theophylline. These findings indicate that AR blockade results in a more rapid and pronounced recruitment of anaerobic metabolism following acute hypoxic exposure, and while rainbow trout and Pacific hagfish show marked differences in their responses to hypoxia, adenosine appears to play an important protective role in both species.

The hypothalamic-pituitary-interrenal axis and the control of food intake in teleost fish.

Although environmental, social and physical stressors have been shown to inhibit food intake and feeding behavior in fish, little is known about the mechanisms that mediate the appetite-suppressing effects of stress. Since the hypothalamic-pituitary-interrenal (HPI) axis is activated in response to most forms of stress in fish, components of this axis may be involved in mediating the food intake reductions elicited by stress. Recent investigations into the brain regulation of food intake in fish have identified several signals with orexigenic and anorexigenic properties. Among these appetite-regulating signals are related neuropeptides that can activate the HPI axis, namely corticotropin-releasing factor (CRF) and urotensin I (UI). Central injections of CRF or UI, or treatments that result in an increase in hypothalamic CRF and UI gene expression, can elicit dose-dependent decreases in food intake that can be reversed by pre-treatment with a CRF-receptor antagonist. Evidence also suggests that cortisol, the end product of HPI activation in most fishes (i.e. Osteichthyes), may be involved in the regulation of food intake. Overall, while elements of the HPI axis may mediate some of the appetite-suppressing effects of stress, it is undetermined how either CRF-related peptides, cortisol, or other elements of the stress response interact with the complex circuitry of the hypothalamic feeding center.

The adrenergic stress response in fish: control of catecholamine storage and release.

In fish, the catecholamine hormones adrenaline and noradrenaline are released into the circulation, from chromaffin cells, during numerous 'stressful' situations. The physiological and biochemical actions of these hormones (the efferent adrenergic response) have been the focus of numerous investigations over the past several decades. However, until recently, few studies have examined aspects involved in controlling/modulating catecholamine storage and release in fish. This review provides a detailed account of the afferent limb of the adrenergic response in fish, from the biosynthesis of catecholamines to the exocytotic release of these hormones from the chromaffin cells. The emphasis is on three particular topics: (1) catecholamine biosynthesis and storage within the chromaffin cells including the different types of chromaffin cells and their varying arrangement amongst species; (2) situations eliciting the secretion of catecholamines (e.g. hypoxia, hypercapnia, chasing); (3) cholinergic and non-cholinergic (i.e. serotonin, adrenocorticotropic hormone, angiotensin, adenosine) control of catecholamine secretion. As such, this review will demonstrate that the control of catecholamine storage and release in fish chromaffin cells is a complex processes involving regulation via numerous hormones, neurotransmitters and second messenger systems.

Extracellular carbonic anhydrase in the dogfish, Squalus acanthias: a role in CO2 excretion.

In Pacific spiny dogfish (Squalus acanthias), plasma CO(2) reactions have access to plasma carbonic anhydrase (CA) and gill membrane-associated CA. The objectives of this study were to characterise the gill membrane-bound CA and investigate whether extracellular CA contributes significantly to CO(2) excretion in dogfish. A subcellular fraction containing membrane-associated CA activity was isolated from dogfish gills and incubated with phosphatidylinositol-specific phospholipase C. This treatment caused significant release of CA activity from its membrane association, a result consistent with identification of the dogfish gill membrane-bound CA as a type IV isozyme. Inhibition constants (K(i)) against acetazolamide and benzolamide were 4.2 and 3.5 nmol L(-1), respectively. Use of a low dose (1.3 mg kg(-1) or 13 micromol L(-1)) of benzolamide to selectively inhibit extracellular CA in vivo caused a significant 30%-60% reduction in the arterial-venous total CO(2) concentration difference, a significant increase in Pco(2) and an acidosis, without affecting blood flow or ventilation. No effect of benzolamide on any measure of CO(2) excretion was detected in rainbow trout (Oncorhynchus mykiss). These results indicate that extracellular CA contributes substantially to CO(2) excretion in the dogfish, an elasmobranch, and confirm that CA is not available to plasma CO(2) reactions in rainbow trout, a teleost.

Distribution and regional stressor-induced regulation of corticotrophin-releasing factor binding protein in rainbow trout (Oncorhynchus mykiss).

The corticotrophin-releasing factor (CRF) system plays a key role in the co-ordination of the physiological response to stress in vertebrates. Although the binding protein (BP) for CRF-related peptides, CRF-BP, is an important player in the many functions of the CRF system, the distribution of CRF-BP and the impact of stressors on its expression in fish are poorly understood. In the present study, we describe the distribution of CRF-BP in the brain and peripheral tissues of rainbow trout (Oncorhynchus mykiss) using a combination of real-time reverse transcriptase-polymerase chain reaction, in situ hybridisation and immunohistochemistry. Our results indicate a widespread and highly localised distribution of CRF-BP in the central nervous system, but do not support a significant peripheral production of the protein. Major expression sites in the brain include the area ventralis telencephali, nucleus preopticus, anterior and lateral tuberal nuclei, and the posterior region of the pituitary pars distalis. We further characterise changes in CRF-BP gene expression in three discrete brain regions after exposure to 8 h and 24 h of social stress or hypoxia. The plasma cortisol concentration in subordinate fish was much higher than in dominant fish and controls, and was indicative of a relatively severe stressor. By contrast, the increase in plasma cortisol concentration in fish exposed to hypoxia was characteristic of the response to a mild stressor. Changes in CRF-BP gene expression were only observed after 24 h of either stressor, and were region-specific. CRF-BP mRNA in the telencephalon increased in both subordinate fish and fish exposed to hypoxia, but CRF-BP in the preoptic area only increased after 24 h of hypoxia exposure. In the hypothalamus, CRF-BP mRNA levels decreased in dominant fish relative to controls after 24 h. Taken together, our results support a diverse role for CRF-BP in the central actions of the fish CRF system, but a negligible role in the peripheral functions of circulating CRF-related peptides. Furthermore, the differential changes in forebrain CRF-BP mRNA appear to occur independently of the hypothalamic-pituitary-inter-renal axis.

Induction of four glutamine synthetase genes in brain of rainbow trout in response to elevated environmental ammonia.

The key strategy for coping with elevated brain ammonia levels in vertebrates is the synthesis of glutamine from ammonia and glutamate, catalyzed by glutamine synthetase (GSase). We hypothesized that all four GSase isoforms (Onmy-GS01-GS04) are expressed in the brain of the ammonia-intolerant rainbow trout Oncorhynchus mykiss and that cerebral GSase is induced during ammonia stress. We measured GSase activity and the mRNA expression of Onmy-GS01-GS04 in fore-, mid- and hindbrain and liver, as well as ammonia concentrations in plasma, liver and brain of fish exposed to 9 or 48 h of 0 (control) or 670 micromol l(-1) NH(4)Cl (75% of the 96 h-LC(50) value). The mRNA of all four GSase isoforms were detected in brain (not liver). After 9 h of NH(4)Cl exposure, brain, liver and plasma ammonia content were elevated by two- to fourfold over control values. Midbrain, hindbrain and liver GSase activities were 1.3- to 1.5-fold higher in ammonia-exposed fish relative to control fish. Onmy-GS01-GS04 mRNA levels in brain (not liver) of ammonia-exposed fish (9 h) were significantly elevated by two- to fourfold over control values. After 48 h of the NH(4)Cl treatment, ammonia content and GSase activity, but not mRNA levels, in all tissues examined remained elevated compared to control fish. Taken together, these findings indicate that all four GSase isoforms are constitutively expressed in trout brain and are inducible under high external ammonia conditions. Moreover, elevation of GSase activities in fore-, mid- and hindbrain in response to environmental ammonia underlines the importance of brain GSase in the ammonia-stress response.

Neuropeptides and the control of food intake in fish.

The brain, particularly the hypothalamus, integrates input from factors that stimulate (orexigenic) and inhibit (anorexigenic) food intake. In fish, the identification of appetite regulators has been achieved by the use of both peptide injections followed by measurements of food intake, and by molecular cloning combined with gene expression studies. Neuropeptide Y (NPY) is the most potent orexigenic factor in fish. Other orexigenic peptides, orexin A and B and galanin, have been found to interact with NPY in the control of food intake in an interdependent and coordinated manner. On the other hand cholecystokinin (CCK), cocaine and amphetamine-regulated transcript (CART), and corticotropin-releasing factor (CRF) are potent anorexigenic factors in fish, the latter being involved in stress-related anorexia. CCK and CART have synergistic effects on food intake and modulate the actions of NPY and orexins. Although leptin has not yet been identified in fish, administration of mammalian leptin inhibits food intake in goldfish. Moreover, leptin induces CCK gene expression in the hypothalamus and its actions are mediated at least in part by CCK. Other orexigenic factors have been identified in teleost fish, including the agouti-related protein (AgRP) and ghrelin. Additional anorexigenic factors include bombesin (or gastrin-releasing peptide), alpha-melanocyte-stimulating hormone (alpha-MSH), tachykinins, and urotensin I. In goldfish, nutritional status can modify the expression of mRNAs encoding a number of these peptides, which provides further evidence for their roles as appetite regulators: (1) brain mRNA expression of CCK, CART, tachykinins, galanin, ghrelin, and NPY undergo peri-prandial variations; and (2) fasting increases the brain mRNA expression of NPY, AgRP, and ghrelin as well as serum ghrelin levels, and decreases the brain mRNA expression of tachykinins, CART, and CCK. This review will provide an overview of recent findings in this field.

Angiotensins stimulate catecholamine release from the chromaffin tissue of the rainbow trout.

Immunohistochemical and pharmacological techniques were utilized to investigate the relationships between angiotensins and catecholamine release from the chromaffin tissue of rainbow trout (Oncorhynchus mykiss). Double labeling with [Asp1, Ile5]angiotensin II-fluorescein isothiocyanate (ANG II-FITC) and anti-dopamine beta-hydroxylase revealed specific ANG II binding sites on chromaffin cells. Injection (1 nmol/kg body wt) of either ANG II-FITC, [Asn1, Val5, Asn9]ANG I, [Asp1, Ile5, His9]ANG I, [Asn1, Val5]ANG II, [Asp1, Val5]ANG II, or [Asp1, Ile5]ANG II elicited catecholamine release from in situ perfusion preparations of the head kidney. Catecholamine release elicited by [Asn1, Val5]ANG II (10(-13) to 10(-7) mol/kg body wt) was dose dependent, and the secretion of epinephrine (Epi) was greater than that of norepinephrine (NE). Relative to the results obtained with the [Asn1, Val5]ANG II treatment (1 nmol/kg body wt), Epi release was 72 and 82% lower in response to injections (1 nmol/kg body wt) of [Asn1, Val5]ANG I [amino acid (AA) positions 1-7] and [Asn1, Val5]ANG I (AA 1-6), respectively. Pretreatment with either losartan (10(-5) M), PD-123319 (10(-5) M), or hexamethonium (10(-3) M) had no effect on [Asn1, Val5]ANG II-elicited catecholamine release. Pretreatment with captopril (10(-4) M) significantly reduced [Asn1, Val5, Asn9]ANG I-elicited Epi and NE release and decreased basal catecholamine release. These results provide direct evidence that angiotensins can elicit catecholamine release from the chromaffin tissue via specific ANG II binding sites and indicate that the synthesis of ANG II may be either local or systemic.

Appetite-suppressing effects of urotensin I and corticotropin-releasing hormone in goldfish (Carassius auratus).

Fish urotensin I (UI), a member of the corticotropin-releasing hormone (CRH) family of peptides, is a potent inhibitor of food intake in mammals, yet the role of UI in the control of food intake in fish is not known. Therefore, to determine the acute effects of UI on appetite relative to those of CRH, goldfish were given intracerebroventricular (i.c.v.) injections of carp/goldfish UI and rat/human CRH (0.2-200 ng/g) and food intake was assessed for a 2-hour period after the injection. UI and CRH both suppressed food intake in a dose-related manner and UI (ED50 = 3.8 ng/g) was significantly more potent than CRH (ED50 = 43.1 ng/g). Pretreatment with the CRH receptor antagonist, alpha-helical CRH(9-41), reversed the reduction in food intake induced by i.c.v. UI and CRH. To assess whether endogenous UI and CRH modulate fish appetite, goldfish were given intraperitoneal implants of the glucocorticoid receptor antagonist, RU-486 (50 and 100 microg/g), or the cortisol synthesis inhibitor, metyrapone (100 and 200 microg/g), and food intake was monitored over the following 72 h. Fish treated with either RU-486 or metyrapone were characterized by a sustained and dose-dependent reduction in food intake. Pretreatment with i.c.v. implants of alpha-helical CRH(9-41) partially reversed the appetite-suppressing effects of RU-486 and metyrapone. In a parallel experiment, the effects of RU-486 (100 microg/g) and metyrapone (200 microg/g) intraperitoneal implants on brain UI and CRH gene expression were assessed. Relative to sham-implanted controls, fish treated with RU-486 or metyrapone had elevated UI mRNA levels in the hypothalamus and CRH mRNA levels in the telencephalon-preoptic brain region. Together, these results suggest that UI is a potent anorectic peptide in the brain of goldfish and that endogenous CRH-related peptides can play a physiological role in the control of fish appetite.

Cardiovascular effects of angiotensin-II-mediated adrenaline release in rainbow trout Oncorhynchus mykiss.

To determine the contribution of plasma catecholamines to the cardiovascular effects of elevated levels of angiotensin II (Ang II) in trout, this study investigated (1) the stimulatory effects of [Asn1-Val5]-Ang II on plasma catecholamine levels, (2) the cardiovascular effects of Ang II with and without alpha-adrenoceptor blockade and (3) the relationship between plasma adrenaline concentrations and their cardiovascular effects. Bolus intravascular injections of Ang II (25-1200 pmol kg-1) elicited dose-dependent (between 75 and 1200 pmol kg-1) increases in plasma adrenaline levels; mean plasma noradrenaline levels only increased in response to a dose of 1200 pmol kg-1. Ang-II-elicited increases in plasma adrenaline levels ranged from 3.3+/-0.3 nmol l-1 for 75 pmol kg-1 Ang II to 125.1+/-40.0 nmol l-1 for 1200 pmol kg-1 Ang II. Injections of Ang II (25-1200 pmol kg-1) also elicited dose-dependent increases in dorsal aortic pressure (PDA), systemic resistance (RS), cardiac output (Q) and stroke volume (Vs). In fish first treated with the alpha -adrenoceptor blocker phenoxybenzamine, Ang II injections elicited a decrease in q_dot and Vs, and the increases in PDA and RS following administration of the 600 and 1200 pmol kg-1 Ang II doses were significantly reduced. Bolus injections of adrenaline (1.8x10(-10) to 1.4x10(-8) mol kg-1) elicited dose-dependent increases in PDA at a plasma adrenaline concentration of 16.5 nmol l-1 and in RS at a plasma adrenaline concentration of 50.5 nmol l-1. Adrenaline injections also elicited increases in Q and Vs at plasma adrenaline concentrations of 50.5 nmol l-1; however, higher plasma adrenaline concentrations were not associated with further increases in either Q or Vs. These results demonstrate that, in vivo, Ang II can act as a potent non-cholinergic secretagogue of humoral adrenaline in trout and that some of the cardiovascular effects of exogenous Ang II can be attributed to increased levels of plasma adrenaline. Our data also indicate that the cardiovascular effects of Ang-II-mediated humoral catecholamines are recruited in a dose-dependent manner and, as such, may require an acute stimulation of the renin-angiotensin system to contribute significantly to the pressor activity of endogenous angiotensins.