Arginine vasotocin impacts chemosensory behavior during social interactions of Anolis carolinensis lizards.

In reptiles, arginine vasotocin (AVT) impacts the performance of and response to visual social signals, but whether AVT also operates within the chemosensory system as arginine vasopressin (AVP) does in mammals is unknown, despite social odors being potent modifiers of competitive and appetitive behavior in reptiles. Here, we ask whether elevated levels of exogenous AVT impact rates of chemical display behavior (e.g. tongue flicks) in adult males, and whether conspecific males or females can chemically discriminate between competitor males based on differing levels of exogenous AVT in green anoles (Anolis carolinensis). We injected wild-caught green anole males with either AVT (AVT-Males) or a vehicle control (CON-Males) solution, then presented treated males with a conspecific stimulus (Intruder-Male or Intruder-Female) and filmed 30-minute interactions. We found that AVT-Males were faster than CON-Males to perform a tongue flick to conspecifics, and faster to chemically display toward Intruder-Females, suggesting AVT increased male interest in available chemical information during social encounters. Intruders performed more lip smack behavior when interacting with AVT-Males than with CON-Males, and Intruder-Males performed more tongue flick behavior when interacting with AVT-Males than with CON-Males, suggesting anoles can discriminate between conspecifics based on exogenous AVT levels. We also found a reduction in Intruder movement behavior when Intruders were paired with AVT-Males. This study provides empirical support for AVT-mediated chemosensory behavior in reptilian social interactions, in a microsmatic lizard species, suggesting the mechanism by which mammalian AVP and non-mammalian AVT mediate chemosensory behavior during social interactions may be evolutionarily conserved.

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

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

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

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

Metabolic status and ghrelin regulate plasma levels and release of ovarian hormones in layer chicks.

The aim of the present study was to examine the role of nutritional status, the metabolic hormone ghrelin and their interrelationships in the control of chicken hormones involved in the regulation of reproduction. For this purpose, we identified the effect of food deprivation, administration of ghrelin 1-18 and their combination on plasma levels of testosterone (T), estradiol (E), arginine-vasotocin (AVT) and growth hormone (GH) as well as the release of these hormones by isolated and cultured ovarian fragments. It was observed that food deprivation reduces plasma T and E and increases plasma AVT and GH levels. Food restriction also reduced the amount of E produced by isolated ovaries, but it did not affect the ovarian secretion of T and AVT. No ovarian GH secretion was detected. Ghrelin administered to ad libitum fed chickens did not affect plasma T and E levels, but it did increase plasma GH and AVT concentrations. Moreover, it partially prevented the effect of food deprivation on plasma E and AVT levels, but not on T or GH levels. Ghrelin administration to control birds promoted ovarian T, but not E or AVT release and reduced T and no other hormonal outputs in birds subjected to food restriction. Our results (1) confirmed the ovarian origin of the main plasma T and E and the extra-ovarian origin of the main blood AVT and GH; (2) showed that food deprivation-induced suppression of reproduction may be caused by suppression of T and E and the promotion of AVT and GH release; (3) suggest the involvement of ghrelin in control chicken E, AVT and GH output; and (4) indicates that ghrelin can either mimic or modify the effect of the intake of low calories on chicken plasma and ovarian hormones, i.e. it can mediate the effect of metabolic state on hormones involved in the control of reproduction.

Changes in plasma angiotensin II, aldosterone, arginine vasotocin, corticosterone, and electrolyte concentrations during acclimation to dry condition and seawater in the crab-eating frog.

The crab-eating frog Fejervarya cancrivora inhabits mangrove swamps and marshes in Southeast Asia. In the present study, circulating angiotensin II (Ang II), aldosterone (Aldo), arginine vasotocin (AVT), and corticosterone (Cort) concentrations as well as various blood parameters were studied under osmotically stressful conditions. Following acclimation to hyperosmotic seawater and dry condition for 5days, body weight was significantly decreased. Under both conditions, plasma Na(+), Cl(-), and urea concentrations, hematocrit values (Ht; blood volume indicator), and osmolality were significantly increased. Dehydration associated with hypovolemic and hyperosmotic states of body fluids was induced during acclimation to hyperosmotic seawater and dry condition in the crab-eating frogs. Ang II, Aldo, AVT, and Cort were maintained within relatively narrow concentration ranges in the control frogs; however, in frogs under dry and hyperosmotic seawater conditions, large variations were observed among individuals in each group. Mean plasma Ang II and Aldo concentrations significantly increased in hyperosmotic seawater-acclimated and desiccated frogs. Although mean plasma AVT concentrations in dehydrated frogs of both the groups were approximately 2.0-3.5 times higher than those in the control frogs, the differences were not significant because of the variation. There was a significant correlation between plasma osmolality and AVT as well as Ang II but not Aldo. A significant correlation was also observed between Ht and AVT as well as Ang II. Plasma Ang II was significantly correlated with plasma Aldo. These results indicate that the crab-eating frogs may exhibit similar physiological responses to both seawater-acclimated and dry conditions. It appears that under dehydrated conditions, osmoregulatory mechanisms participate in stabilization of the situation. The renin-angiotensin system may have pivotal roles in body fluid regulation under volemic and osmotic stress in the Fejervarya species with unique osmoregulation.

Differential and reproductive stage-dependent regulation of vasotocin secretion by catecholamines in the catfish Heteropneustes fossilis.

Vasotocin (VT) is the basic nonapeptide hormone secreted by the neurohypophysis of non-mammalian vertebrates and is involved in the regulation of osmoregulation, metabolism, cardiovascular function, reproduction and behaviour. Among the reproductive function, VT is specifically implicated in final oocyte maturation, ovulation, oviposition/parturition in teleosts, amphibians, reptiles, and birds. The central catecholaminergic system is involved in the regulation of pituitary hormone secretion including gonadotropin, and mediates also changes in environmental photoperiod and temperature. The close apposition of the VT and catecholaminergic systems in the preoptic area of the hypothalamus signifies a strong possibility of their functional interaction. The aim of the present study was to investigate the effects of exogenously administered catecholamines on VT secretion in two different reproductive phases of female catfish Heteropneustes fossilis. For this, the catecholamine precursor L-dihydroxyphenylalanine (L-DOPA) and catecholamines (dopamine-DA, norepinephrine-NE, and epinephrine-E) were intraperitoneally injected in normal catfish and/or along with α-methylparatyrosine (α-MPT, a tyrosine hydroxylase inhibitor). Brain and plasma VT levels were measured by specific ELISA, 24h post injection. Both L-DOPA and DA inhibited brain and plasma VT levels in a concentration-dependent manner in preparatory and prespawning phases. In contrast, NE elicited dose-dependent effects: the lowest dose (0.5 ng/g body mass, BM) was ineffective, the median dose (1 ng/g BM) stimulated, and the high doses (10 and 100 ng/g BM) inhibited VT levels. E stimulated VT levels dose-dependently. A single injection of α-MPT (250 µg/g BM) strongly inhibited VT when given alone and enhanced the inhibitory effects of L-DOPA and DA in the combination groups. The α-MPT inhibition of VT was significantly reduced by the injection of NE (5 ng/g BM) and was restored or elevated by E. When the adrenergic neurotransmitters were given together with α-MPT, the inhibitory effect of the latter was abolished and VT levels were significantly elevated. Thus, the present data indicate that the physiological changes in VT is differentially regulated by the catecholamines (DA inhibits and NE/E stimulates VT).

Skin surface temperature of broiler chickens is correlated to body core temperature and is indicative of their thermoregulatory status.

Extreme thermal conditions may dramatically affect the performance of broilers and other domestic animals, thereby impairing animal welfare and causing economic losses. Although body core temperature is the parameter that best reflects a bird's thermal status, practical and physiological obstacles make it irrelevant as a source of information on the thermal status of commercial flocks. Advances in the technology of infrared thermal imaging have enabled highly accurate, noncontact, and noninvasive measurements of skin surface temperature. Providing that skin surface temperature correlates with body temperature, this technology could enable acquisition of reliable information on the thermal status of animals, thereby improving diagnoses of environmental stress in a flock. This study of broiler chickens found a strong positive correlation between body core temperature and facial surface temperature, as recorded by infrared thermal imaging. The correlation was equally strong at all ages from 8 to 36 d during exposure to acute heat stress with or without proper ventilation and after acclimation to chronic heat exposure. A similar correlation was found by measurements in commercial flocks of broilers. Measurements of blood plasma concentrations of corticosterone, thyroid hormones, and arginine vasotocin confirmed that metabolic activity was low after acclimation to chronic exposure to heat, whereas ventilation was at least as efficient as acclimation in reducing thermal stress but did not impair metabolism. In light of these novel results, commercial benefits of infrared thermal imaging technology are suggested, especially in climate control for commercial poultry flocks. The application of this technique to other domestic animals should be investigated in future experiments.

Diurnal rhythms in hypothalamic/pituitary AVT synthesis and secretion in rainbow trout: evidence for a circadian regulation.

Arginine vasotocin (AVT) and isotocin (IT) are two neurohypophysial peptide hormones for which a role in adaptation to environmental changes has been suggested in fish. In teleosts, there are only a few available studies about circadian changes of AVT and IT levels, and a role of those peptides in the circadian system has been mainly suggested on the basis of the role of the homologous hormone AVP in mammals. Herein, we evaluated the diurnal rhythms in plasma AVT, pituitary AVT and IT content and the hypothalamic pro-vasotocin (pro-VT) expression in rainbow trout kept under a natural photoperiod, as well as their persistence in constant darkness as a tool for defining circadian dependence. Trout kept under a natural light cycle showed clear diurnal rhythms in both circulating and pituitary AVT levels with peak values around the last hours of the light phase. Hypothalamic pro-VT mRNA was also rhythmically expressed with similar peak characteristics. These rhythms persisted in fish kept under constant darkness for nearly two consecutive days, although peaks were progressively attenuated and phase-advanced. An IT rhythm was also found in pituitary of the trout maintained under a natural photoperiod, but not in those kept under continuous darkness. These results suggest that rhythms of hypothalamic AVT synthesis might be regulated by endogenous circadian mechanisms, and these rhythms contribute to maintain a similar fluctuation in pituitary AVT secretion into the blood. A potential role for AVT in the circadian and seasonal time-keeping system of teleost fish, either as a component of the neural machinery that participates in the adaptation to cyclic environmental changes, or as a circadian/seasonal output signal, is also discussed.

Calcitonin receptor binding in the hen neurohypophysis before and after oviposition.

To demonstrate the presence of a receptor for calcitonin (CT) in the hen neurohypophysis and to estimate the time of action of CT on the neurohypophysis during the oviposition cycle in relation to arginine vasotocin (AVT) release, binding of (125)I-labeled chicken CT in plasma membrane fractions of the hen neurohypophysis was measured by the use of a radioligand binding assay. The binding specificity, reversibility, high affinity, and limited capacity are characteristics of a CT receptor. Therefore, it was elucidated that the CT receptor might exist in the plasma membrane of the neurohypophysis of hens. The binding affinity of CT receptor increased at 30 min before oviposition and the binding capacity was decreased at 15 min before oviposition. However, no change was found in non-laying hens during a 24-h period. Such changes in the CT receptor binding were found at 10 min after an i.v. injection of chicken CT into non-laying hens with an increase in the blood level of AVT. The changes in the binding affinity and capacity of CT receptor of the neurohypophysis may be related to AVT release partly at oviposition time in the hen.

Quantitative determination of oxytocin receptor antagonist atosiban in rat plasma by liquid chromatography-tandem mass spectrometry.

A kinetic study of atosiban was conducted following repeated intravenous administration in Wistar rats. Sample analysis was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) following full validation of an in-house method. Eptifibatide, a cyclic peptide, was used as an internal standard (IS). The analyte and internal standard were extracted using solid phase extraction (SPE) method. Chromatographic separation was carried out using an ACE C18 5 microm 50 mm x 4.6 mm column with gradient elution. Mass spectrometric detection was performed using TSQ Quantum ultra AM. The lower limit of quantification was 0.01 microg/ml when 100 microl rat plasma was used. Plasma concentrations of atosiban were measured at 0 (pre-dose), 2, 15, 30, 45, 60, 120 min at the dosage levels of 0.125 mg/kg (low dose), 0.250 mg/kg (mid dose), and 0.500 mg/kg (high dose), respectively. Atosiban plasma concentration measured at Day 1 showed mean peak atosiban concentration (C(max)) 0.40, 0.57, 1.95 microg/ml for low, mid and high dose treated animals and mean peak concentration on Day 28 was 0.41, 0.88, 1.31microg/ml on Day 28 for low, mid and high dose treated animals.

Effects of hCG and ovarian steroid hormones on vasotocin levels in the female catfish Heteropneustes fossilis.

Effects of hCG, ovariectomy and estradiol replacement on brain, plasma and/or ovarian vasotocin in vivo, and estradiol, progesterone, 17alpha, 20beta-hydroxy-4-pregnen-3-one and hCG on ovarian vasotocin in vitro were investigated in the catfish. A 100IU/fish of hCG induced ovulation and elicited both periovulatory and post-ovulatory changes in vasotocin concentrations with a significant increase up to 8h in the brain and up to 16h in both plasma and ovary. After stripping the fish at 16h, the peptide concentration decreased significantly with time, up to 4 days. Ovariectomy in early pre-spawning phase resulted in a duration-dependent significant reduction of both brain and plasma vasotocin. Estradiol replacement in 3-week ovariectomized fish produced dosage-dependent biphasic effects: the lower dosage (0.1microg/g) restored the vasotocin level while the higher dosage (0.5microg/g) decreased it significantly below the control level. In vitro incubation of ovarian tissues with estradiol produced season-dependent effects on vasotocin. The incubation of pre-vitellogenic ovarian pieces with estradiol (1, 10, and 100ng/ml) elevated vasotocin level in a dose- and duration-dependent manner while that of post-vitellogenic follicles resulted in a significant decrease. The incubation of intact post-vitellogenic follicles or follicular envelope with progesterone and 17alpha, 20beta-hydroxy-4-pregnen-3-one (1microg/ml) or hCG (20IU/ml) for 8 and 16h significantly increased vasotocin in a duration-dependent manner. The results show that both gonadotropin and ovarian steroids modulate vasotocin titer, which may influence follicular growth, ovulation and spawning in the catfish.

Immunocytochemical localization, HPLC characterization, and seasonal dynamics of vasotocin in the brain, blood plasma and gonads of the catfish Heteropneustes fossilis.

Immunocytochemical distribution and dynamics of vasotocin (VT) were studied in the air-breathing catfish Heteropneustes fossilis in relation to the reproductive cycle. Vasotocin was localized in the brain and ovary by streptavidin-biotin immunocytochemistry. The immunoreactivity was found throughout the hypothalamo-hypophysial neurosecretory system consisting of the magnocellular and parvocellular neurons of the nucleus preopticus, neurosecretory axonal tract and neurohypophysis (NH). The VT neurons showed seasonal changes; they were numerically less in resting phase but increased during the recrudescent phase. The neurons were hypertrophied and degranulated in pre-spawning phase and heavily degranulated and vacuolated in spawning phase. In the NH, the density of VT fibers increased up to the pre-spawning phase and decreased thereafter. In the ovary, VT immunoreactivity was noticed in the follicular layer and varied with the growth of the follicles. Vasotocin was characterized and quantified by a high performance liquid chromatography with UV detection method in the brain, plasma and ovary. Brain and plasma VT concentrations were also assayed with an EIA method, which was more sensitive than the HPLC method with values about 2-fold higher. Vasotocin levels showed significant seasonal and sexual differences with higher concentrations in females in the recrudescent (preparative, pre-spawning and spawning) phase. Brain VT recorded the highest concentration in the preparative phase (both sexes) while plasma (both sexes) and ovarian VT in the spawning phase. The ovarian concentration of VT was 15- and 25-fold higher in the pre-spawning and spawning phases (when expressed per mg protein), respectively, than plasma but lower than brain levels. In testis, VT concentration was relatively low and apparently did not show any significant seasonal variation. The seasonal activity patterns and gonadal distribution of VT indicate a reproductive function of the peptide.

Effect of melatonin on salt gland and kidney function of gulls, Larus glaucescens.

This study examined effects of exogenous melatonin on osmoregulatory hormones and water and sodium secretion by salt glands and excretion via the kidneys of Glaucous-winged gulls (Larus glaucescens). Six saline acclimated gulls were injected with inulin and paraminohippuric acid and then infused with 500 mM NaCl to stimulate salt gland secretion. Each bird was given infusions of NaCl alone and NaCl plus melatonin. Experiments were made one week apart in a randomized order. A large blood sample (to measure osmoregulatory hormones) was taken before infusion, at secretion, and at the end of infusion. A small blood sample was taken at the midpoint of each of six 10 min sequential collections of salt gland secretion and urine. Melatonin tended to increase plasma sodium concentration, did decrease plasma osmolality, but did not affect potassium concentration. Melatonin did not affect salt gland secretion rate or concentration nor renal plasma flow or glomerular filtration. Melatonin increased urine flow rate, tended to increase urine sodium concentration, and did decrease urine potassium concentration. Combined renal and extrarenal sodium excretion was greater during MT treatment. During NaCl infusion, angiotensin II increased, aldosterone decreased, and arginine vasotocin remained unchanged. Melatonin did not affect these responses. These data suggest an osmoregulatory role for melatonin in birds with salt glands.

Effect of hypernatraemia and the neurohypophysial peptide, arginine vasotocin (AVT) on behavioural thermoregulation in the agamid lizard, Ctenophorus ornatus.

Hypernatraemia induced by chronic injections of sodium chloride provokes thermal depression in the agamid lizard, Ctenophorus (formerly Amphibolurus) ornatus, with a fall of two degrees Celsius in the mean body temperature selected behaviourally in a photo-thermal gradient. The placement of an electrolytic lesion in the base of the hypothalamus, designed to eliminate secretion of the neuropeptide arginine vasotocin (AVT), did not affect the lizards' thermoregulatory behaviour and their Preferred Body Temperature (PBT) was not significantly different from that of unoperated controls. Saline loading, however, did not induce thermal depression in these tract-operated individuals and their PBT was significantly higher than that of salt-loaded intact individuals. When AVT was injected into operated, salt-loaded, animals, however, thermal depression was observed, supporting the hypothesis that thermal depression brought about by hypernatraemia is mediated through the action of AVT. AVT similarly significantly depressed the PBT of injected intact individuals by 3.2 degrees C when compared with hydrated controls. Immunostaining for AVT confirmed that the lesions placed in the region of the median eminence virtually eliminated AVT located in the neurohypophysial tract, and the pars nervosa. This is the first report of an effect of this peptide on behavioural thermoregulation in a lizard.

Influences of sex and saline intake on diurnal changes in plasma melatonin and osmoregulatory hormones of Pekin ducks (Anas platyrhynchos).

Melatonin (MT) inhibits salt gland secretion of Pekin ducks [Ching, A.C.T., Hughes, M.R., Poon, A.M.S., Pang, S.F., 1999. Melatonin receptors and melatonin inhibition of duck salt gland secretion. Gen. Comp. Endocrinol. 116, 229-240]. The present study examined simultaneous diurnal changes in plasma concentrations of MT and four osmoregulatory hormones of Pekin ducks to assess the possible interactions among these hormones that might contribute to inhibition of extrarenal salt secretion. The study compared diurnal changes in freshwater (FW) and saline-acclimated (SA) male and female ducks. We hypothesized plasma concentrations of osmoregulatory hormones: (1) differ between sexes (partially accepted); (2) vary diurnally (accepted, influenced by sex and treatment); (3) are correlated with MT (partially accepted); and their diurnal variation is affected by (4) sex (partially accepted); and (5) saline acclimation (partially accepted). We compared body mass, plasma osmolality (Osm(pl)), plasma volume, concentrations of plasma ions, and arginine vasotocin (AVT), angiotensin II (AII), prolactin, (PRL), and aldosterone (ALDO), in relation to plasma melatonin (MT). At night body mass increased in FW ducks and decreased in SA ducks. This likely reflects the higher plasma PRL concentration of female ducks and its middark increase only in SA ducks. Plasma volume increased at night in all but FW males. Plasma sodium and osmolality were higher in SA ducks and did not vary diurnally in either FW or SA ducks. Midday MT did not differ between males and females or between FW and SA ducks; at middark, all ducks, except FW females, increased MT. Midday AVT was higher in SA ducks. Only FW ducks increased AVT at middark. Changes in AVT and MT were positively correlated in males and negatively correlated in females. Males had higher AII and, at night, AII increased in SA ducks and decreased in FW ducks. AVT and AII were negatively correlated. ALDO was highly correlated with AII and unaffected by sex, saline, or time. At night, only AVT was elevated in FW ducks, while MT, AII, and PRL were elevated in SA ducks. FW and SA ducks appear to follow different, but equally effective, hormonal strategies to maintain osmotic homeostasis at night. We conclude that the effect of MT on salt gland secretion is probably a direct effect. These may be influenced by gender.

Water balance of field-excavated aestivating Australian desert frogs, the cocoon-forming Neobatrachus aquilonius and the non-cocooning Notaden nichollsi (Amphibia: Myobatrachidae).

Burrowed aestivating frogs of the cocoon-forming species Neobatrachus aquilonius and the non-cocooning species Notaden nichollsi were excavated in the Gibson Desert of central Australia. Their hydration state (osmotic pressure of the plasma and urine) was compared to the moisture content and water potential of the surrounding soil. The non-cocooning N. nichollsi was consistently found in sand dunes. While this sand had favourable water potential properties for buried frogs, the considerable spatial and temporal variation in sand moisture meant that frogs were not always in positive water balance with respect to the surrounding soil. The cocoon-forming N. aquilonius was excavated from two distinct habitat types, a claypan in which frogs had a well-formed cocoon and a dune swale where frogs did not have a cocoon. Cocoons of excavated frogs ranged in thickness from 19.4 microm to 55.61 microm and consisted of 81-229 layers. Cocooned claypan N. aquilonius were nearing exhaustion of their bladder water reserves and had a urine osmolality approaching that of the plasma. By contrast, non-cocooned N. aquilonius from the dune swale were fully hydrated, although soil moisture levels were not as high as calculated to be necessary to maintain water balance. Both species had similar plasma arginine vasotocin (AVT) concentrations ranging from 9.4 to 164 pg ml(-1), except for one cocooned N. aquilonius with a higher concentration of 394 pg ml(-1). For both species, AVT showed no relationship with plasma osmolality over the lower range of plasma osmolalities but was appreciably increased at the highest osmolality recorded. This study provides the first evidence that cocoon formation following burrowing is not obligatory in species that are capable of doing so, but that cocoon formation occurs when soil water conditions are more desiccating than for non-cocooned frogs.

Physiological and hormonal control of thermal depression in the tiger snake, Notechis scutatus.

Plasma sodium concentrations in field-caught Western tiger snakes, Notechis scutatus, from semi-arid Carnac Island (CI) varied seasonally, with snakes exhibiting significant hypernatraemia during summer and normal concentrations following autumn rain. In contrast, field-caught tiger snakes from a perennial fresh-water swamp (Herdsman Lake, HL) exhibited no significant increase in plasma sodium concentrations during summer. Laboratory-induced hypernatraemia caused thermal depression in both populations; there was a weak negative relationship between plasma sodium concentration and temperature selection that was significant for CI snakes. Hypernatraemia significantly elevated circulating concentrations of the neuropeptide arginine vasotocin (AVT) in both CI and HL snakes. CI snakes injected with a physiological dosage of AVT also evidenced thermal depression. Despite the positive correlation between AVT and both plasma sodium concentration and osmolality for laboratory snakes, field samples from CI snakes indicate that circulating levels of AVT may be influenced more by plasma osmolality than sodium levels. The data suggest that, in CI snakes, chronic dehydration in the field leads to hypernatraemia which may lead to elevated levels of AVT if plasma osmolality also increases. This will in turn invoke a depression in thermal behaviour that may improve the water economy and survival of snakes on semi-arid CI. Although HL snakes do not experience seasonal dehydration, physiological changes away from the stable homeostatic state appear to prompt the same behavioural shifts, illustrating the intrinsic nature of the thermal behaviour in different populations of the same species of snake.

Kidney function and the role of arginine vasotocin in three agamid lizards from habitats of differing aridity in Western Australia.

Western Australian agamid lizards are diverse and inhabit mesic to very arid areas of the state. Although reptilian kidneys are unable to elaborate hyperosmotic urine, we hypothesised that the renal system of lizards inhabiting arid areas would display an enhanced ability to conserve water under the control of the antidiuretic peptide hormone, arginine vasotocin (AVT). To examine this, the renal physiological and endocrine responses to osmotic challenge in three closely-related Australian agamid lizards inhabiting arid, semi-arid, and mesic environments were studied. The species studied were Pogona minor (mesic), Ctenophorus salinarum (semi-arid), and Ctenophorus nuchalis (arid). Circulating AVT was assayed and renal variables such as glomerular filtration rate (GFR), urine flow rate (V), and fractional reabsorption of filtrate FRH2O were measured in response to hypernatraemia, water load, and dehydration. Hypernatraemia and dehydration induced antidiuresis in all three species through similar mechanisms involving both glomerular and tubular responses. However, in salt-loaded P. minor the response was largely glomerular in nature, as FRH2O did not increase relative to the hydrated condition. The magnitude of the antidiuretic response was also greater in P. minor, indicating a greater sensitivity to osmotic challenge. Plasma concentrations of AVT were significantly correlated with FRH2O in P. minor (r2=0.38, P=0.025), but with GFR in C. nuchalis (r2=0.16, P=0.041). We found that the control and mechanisms of renal function among these lizards were largely similar, and there was little support for the hypothesis that arid lizards possess physiological adaptations not present in closely-related mesic lizards. Yet, differences remain in their response to hypernatraemia which may reflect the aridity of their different environments, or their varying habits.

The effects of freshwater to seawater transfer on circulating levels of angiotensin II, C-type natriuretic peptide and arginine vasotocin in the euryhaline elasmobranch, Carcharhinus leucas.

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.

Altered plasma and pituitary arginine vasotocin and hypothalamic provasotocin expression in flounder (Platichthys flesus) following hypertonic challenge and distribution of vasotocin receptors within the kidney.

Plasma AVT concentration, pituitary AVT content, hypothalamic provasotocin mRNA expression and other osmoregulatory parameters were measured in euryhaline flounder 4, 8, and 24 h after the hypertonic challenge of transfer from fresh water (FW) to seawater (SW). Osmolality and the concentration of major plasma ions, sodium and chloride, were significantly higher in fish transferred to SW by comparison with time matched controls, an effect evident within 4 h. By comparison with time matched controls, pituitary store of AVT was lower while plasma AVT concentration was higher 8 and 24 h after transfer to SW. Higher provasotocin mRNA expression in the hypothalamus was also seen at 4 and 8 h in flounder transferred from FW to SW compared with time matched controls. The lower pituitary store and higher circulating levels imply substantial AVT secretion occurs in the early phase response to this hypertonic challenge. Changes in the regulation of AVT synthesis and secretion appeared quickly following movement to SW, consistent with the rapid osmoregulatory response, including reduced urine production that fish require to accommodate the dehydrative water losses and salt loading on exposure to the new hyperosmotic environment. qPCR measures of whole kidney vasotocin receptor mRNA expression indicated similar levels in SW and FW. Immunohistochemistry for the vasotocin receptor in flounder kidney showed localisation on the afferent and efferent arterioles of the glomerulus and on the capillary bed that extends from the efferent arteriole to the smooth muscle surrounding the collecting duct. Localisation of the vasotocin receptor was comparable in SW and FW fish.