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PubMed1994

Structure, function, and phylogeny of [Arg8]vasotocin receptors from teleost fish and toad.

[Arg8]Vasotocin (AVT) is considered to be the most primitive known vertebrate neurohypophyseal peptide of the vasopressin/oxytocin hormone family and may thus be ancestral to all the other vertebrate peptide hormones. The molecular evolution of the corresponding receptor family has now been studied by cloning an AVT receptor, consisting of 435 amino acid residues, from the teleost fish, the white sucker Catostomus commersoni. Frog oocytes injected with the AVT receptor-encoding cRNA respond to the application of AVT, but not to its structural and functional counterpart isotocin, by an induction of membrane chloride currents indicating the coupling of the AVT receptor to the inositol phosphate/calcium pathway. The pharmacological properties of the expressed AVT receptor show that it represents, or is closely related to, an ancestral nonapeptide receptor: oxytocin, aspargtocin, mesotocin, and vasopressin activated the receptor, but other members of the vasopressin/oxytocin family tested showed little or no potency; antagonists of the mammalian vasopressin V1 and oxytocin receptors blocked the AVT response. Comparison of AVT receptor sequences spanning transmembrane domains two to five, deduced by cloning cDNAs from the Pacific salmon Oncorhynchus kisutch, the cave-dwelling fish Astyanax fasciatus, and the anuran Xenopus laevis, with those of their mammalian counterparts emphasizes amino acid residues that are involved in hormone binding. The presence of a 5.0-kb transcript in various teleost tissues (pituitary, liver, gills, swim bladder, and lateral line) points to a physiological role for the fish AVT receptor in metabolic, osmoregulatory, and sensory processes.

Amino Acid Sequence↗

PubMed1997

Australian lungfish neurohypophysial hormone genes encode vasotocin and [Phe2]mesotocin precursors homologous to tetrapod-type precursors.

In view of the well-established role of neurohypophysial hormones in osmoregulation of terrestrial vertebrates, lungfishes are a key group for study of the molecular and functional evolution of the hypothalamo-neurohypophysial system. Here we report on the primary structure of the precursors encoding vasotocin (VT) and [Phe2]mesotocin ([Phe2]MT) of the Australian lungfish, Neoceratodus forsteri. Genomic sequence analysis and Northern blot analysis confirmed that [Phe2]MT is a native oxytocin family peptide in the Australian lungfish, although it has been reported that the lungfish neurohypophysis contains MT. The VT precursor consists of a signal peptide, VT, that is connected to a neurophysin by a Gly-Lys-Arg sequence, and a copeptin moiety that includes a Leu-rich core segment and a glycosylation site. In contrast, the [Phe2]MT precursor does not contain a copeptin moiety. These structural features of the lungfish precursors are consistent with those in tetrapods, but different from those in teleosts where both VT and isotocin precursors contain a copeptin-like moiety without a glycosylation site at the carboxyl terminals of their neurophysins. Comparison of the exon/intron organization also supports homology of the lungfish [Phe2]MT gene with tetrapod oxytocin/MT genes, rather than with teleost isotocin genes. Moreover, molecular phylogenetic analysis shows that neurohypophysial hormone genes of the lungfish are closely related to those of the toad. The present results along with previous morphological findings indicate that the hypothalamo-neurohypophysial system of the lungfish has evolved along the tetrapod lineage, whereas the teleosts form a separate lineage, both within the class Osteichthyes.

Amino Acid Sequence↗

PubMed1984

Changes in the plasma concentration of immunoreactive arginine vasotocin during oviposition in the domestic fowl.

The plasma concentrations of immunoreactive arginine vasotocin (AVT) were measured during oviposition and shortly before ovulation of the first egg (Cl) of a clutch. Immunoreactive AVT was determined on bentonite extracts of 0.5 ml plasma samples using the method of Rosenbloom and Fisher (1974). The R-70 antiserum used to measure AVT cross reacted with arginine vasopressin (AVP), however the fowl pituitary does not synthesise AVP. Over a period of 10 to 90 min before oviposition the plasma AVT concentration was about 20 pg/ml; during oviposition it increased four-fold. Measurements made at frequent intervals showed that plasma AVT concentration increased 5 to 6 min before oviposition, reached a peak during oviposition itself and decreased rapidly in the following 5 to 6 min. The surge in plasma AVT occurred on average 48 min before Cl ovulation.

Animals↗

PubMed1986

Body temperature and plasma arginine vasotocin in fowls adapted to high- and low-NaCl diets.

The effects of high- and low-NaCl diets on body temperature, plasma osmolality, plasma electrolytes and arginine vasotocin (AVT) were studied in the domestic fowl. Body temperature as a function of time increased or decreased significantly in the high- and low-NaCl groups respectively. Plasma osmolality, sodium and chloride concentrations and AVT concentration were significantly higher in the high-NaCl adapted group. An overall significant correlation was found between body temperature and plasma sodium-to-calcium ratio (r = 0.62). These findings support the concept of an altered hypothalamic thermoregulatory set-point, induced by changes in plasma sodium-to-calcium ratio.

Animals↗

PubMed1988

Physiological effects of arginine vasotocin and mesotocin in cockerels.

1. The effects of continuous infusion of 0.1, 1.0 and 10.0 mU/min/kg body weight of arginine vasotocin (AVT) or mesotocin (MT) on cardiovascular and thermoregulatory responses, on plasma osmolality and ionic composition and on plasma concentrations of AVT, MT, prolactin and aldosterone, were investigated in conscious White Leghorn cockerels. 2. Neither of the peptides, at any dose, affected cardiovascular functions, plasma ions and osmolality. Infusion of MT at the rate of 10 mU/min/kg body weight increased respiratory rate. Both peptides at doses of 1 and 10 mU/min/kg reduced the temperatures of the comb and shank but had no effect on the skin and cloaca. 3. Doses of 0.1 and 1.0 mU MT/min/kg reduced plasma aldosterone and at 10 mU/min/kg increased plasma AVT. At any given dose MT had no effect on plasma prolactin. AVT at 0.1 and 1.0 mU/min/kg of AVT reduced plasma MT. AVT at 1.0 mU/min/kg increased plasma prolactin and at 10 mU/min/kg reduced plasma aldosterone. 4. During saline infusion, plasma MT was positively correlated with plasma AVT and negatively correlated with respiratory rate and cloacal temperature. Plasma AVT showed a positive correlation with plasma MT and aldosterone and a negative correlation with respiratory rate and skin temperature. 5. During saline infusion, there was no significant correlation between cardiovascular functions, or plasma osmolality and ionic composition and plasma MT or AVT. 6. The present study suggests that interrelationships between circulating concentrations of AVT and MT do exist and that AVT affects aldosterone secretion. These neurohypophysical peptides are involved in thermoregulation.

Aldosterone↗

PubMed1990

Plasma concentrations of arginine vasotocin and mesotocin following pentobarbital anaesthesia and carotid cannulation in domestic fowl.

1. Plasma concentrations of arginine vasotocin (AVT) and mesotocin (MT), arterial pH and pCO2, body temperature, respiratory rate, heart rate, mean arterial blood pressure, packed cell volume and plasma osmolality were monitored in birds for 28 h following anaesthesia and carotid cannulation. 2. Plasma MT concentration decreased after 6 h of surgical recovery but there were no differences between 6 and 28 h of recovery. 3. In contrast, plasma AVT concentration was raised only after 27 h, but no differences were observed between 3 and 28 h of recovery. 4. Anaesthesia and/or carotid cannulation apparently suppressed AVT and enhanced MT release, but no significant change in either hormone was observed after 6 h of surgical recovery under the conditions specified in this study.

Anesthesia↗

PubMed1990

Arginine vasotocin and mesotocin in the anterior hypothalamus, neurohypophysis, proventriculus and plasma of White Leghorn cockerels, during dehydration.

1. The effect of 96 hrs of water deprivation on plasma electrolytes, osmolarity, arginine vasotocin (AVT), mesotocin (MT), and on AVT and MT content in the neurohypophysis, anterior hypothalamic area (AHA) and proventriculus, was studied at 24 hrs intervals, in adult White Leghorn cockerels. 2. Plasma AVT increased three fold during the first 24 hrs but there was no further change during the next 48 hrs. In the last 24 hrs, plasma AVT decreased in about 25%. Plasma MT did not change during the entire period of dehydration. 3. Plasma sodium and osmolarity gradually increased during that time. 4. Neurohypophysial AVT content was depleted by 95% during the period of dehydration while MT content did not change. 5. In the AHA there was no change in AVT levels during dehydration while the levels of MT increased while in the proventriculus there was no change in either AVT or MT levels. 6. For the data collected during the entire experimental period, no correlation was found between plasma osmolarity and plasma AVT, but there was a highly significant negative correlation between plasma osmolarity and neurohypophysial AVT content. 7. It may be suggested that the depletion in AVT content in the neurohypophysis during progressive water deprivation resulted in an insufficient level of AVT in circulation to enable the cockerels to counter the dehydration. This may explain the death of those cockerels which were dehydrated for a further 24 hr period.

Animals↗

PubMed1990

Effects of sex and gonadal steroids on arginine vasotocin and mesotocin in the pineal gland and neurohypophysis of White Leghorn fowls.

1. Pineal and neurohypophysial arginine vasotocin (AVT) and mesotocin (MT) were measured in White Leghorn hens, cockerels and castrated males treated with either testosterone propionate (TP) or oestradiol benzoate (EB) (n = 10/group). The lighting regimen was 14 h light: 10 h dark, supplied by natural diffused sunlight and incandescent bulbs. 2. Both AVT and MT were detected in the pineal gland of all the chickens. 3. There was no significant effect of either sex or treatments on pineal MT. 4. Females had about 4 times more pineal AVT than males, regardless of their treatment. There was no effect of the treatments on pineal AVT in the males. 5. No sexual difference in neurohypophysial AVT was detected, but the neurohypophysis of the castrated males treated with EB contained less AVT than the neurohypophysis of the intact males. 6. Intact males had about twice as much MT in the neurohypophysis as females. Castrated males treated with either TP or EB had similar concentrations of neurohypophysial MT, which were lower than that of the intact males, but higher than that of the females.

Animals↗

PubMed1991

Effect of intrahypothalamic injections of norepinephrine and serotonin on plasma arginine vasotocin and mesotocin in the White Leghorn cockerel.

1. Saline (10 microliters), norepinephrine (NE) and Serotonin (5-HT), 500 nmol each, were injected into the anterior third ventricle (A3V; n = 7) or the posterior third ventricle (P3V; n = 11) of ananesthetised, unrestrained White Leghorn cockerels. Plasma arginine vasotocin (AVT) and mesotocin (MT) were measured 20, 60 and 120 min after injection. 2. Injection of NE into both the A3V and P3V had no significant effect on either plasma AVT or plasma MT at any of the sampling times. 3. Administration of 5-HT into the A3V significantly increased plasma MT about two-fold 20 min following injection. At 120 min time, plasma MT returned to normal. 4. In P3V birds, 5-HT had no effect on plasma MT in the first 20 min, but a significant increase in plasma MT occurred 60 to 120 min after injection. The magnitude of the response was lower than in the A3V cockerels. 5. Plasma AVT was not affected by 5-HT administration into the A3V at any of the sampling times, but 5-HT administration into the P3V caused significant rises in plasma AVT at 120 min. 6. Serotonergic, but not noradrenergic, induction of neurohypophysial peptide secretion was demonstrated.

Animals↗

PubMed1981

The pineal does not concentrate arginine vasotocin from biological fluids in vivo.

The ability of the pineal gland to concentrate circulating arginine vasotocin (AVT) from biological fluids in vivo was examined in rats. Intraventricular and intracisternal injections of AVT failed to elevate net pineal AVT content, even though CSF AVT levels were greatly increased by these treatments. Intravenous injection of AVT did elicit a light increase in pineal AVT, but plasma peptide levels wee elevated enormously. Thus, it seems unlikely that an active uptake process for AVT functions in this gland.

Animals↗

PubMed1995

Enzyme linked immunosorbent assay for the neurohypophyseal hormones arginine vasotocin and isotocin.

A competitive enzyme linked immunosorbent assay (ELISA) using new polyclonal antibodies was developed for the first time for each of two neurohypophyseal hormones: arginine vasotocin (AVT, ubiquitous in vertebrates) and isotocin (IT, restricted to teleost fish). The antibodies obtained were highly specific, showed no cross-reaction between the two peptides and were able to suppress the activity of the peptides in in vitro bioassays. An original feature of the assays was the covalent binding of the peptidic antigen to Covalink microplates. Competition was thereafter made between this bound antigen and the antigen in samples, for a fixed amount of the relevant antibody. Revelation used peroxidase-labeled antibodies. These ELISAs were sensitive enough to detect 1 ng/ml and 0.1 ng/ml for AVT and IT respectively. Moreover, for the first time, the two hormones were measured separately, each in the presence of the other, and shown to exist as circulating hormones in fish.

Amino Acid Sequence↗

PubMed2001

Water deprivation and circadian changes in plasma arginine vasotocin and mesotocin in the domestic hen (Gallus domesticus).

Possible circadian variations in plasma levels of arginine vasotocin (AVT) and mesotocin (MT) were assessed in domestic hens (Gallus domesticus) under a 12h:12h light-dark (LD) schedule. Blood samples were taken at 4h intervals, and neurohypophyseal hormone levels were determined by radioimmunoassay. Marked circadian changes in both AVT and MT were observed in hens provided free access to water. Minimal and maximal AVT levels occurred at 08:00 and 20:00, respectively. Minimal MT levels occurred at 20:00, whereas maximal MT levels occurred over a broad time period of 04:00 to 12:00. In water-deprived hens, plasma AVT levels were elevated at each time point, and the circadian variations in plasma AVT and MT levels were attenuated. These results demonstrate that rhythmicity in neurohypophyseal function in a lower vertebrate species, like that in mammals, is disrupted by osmotic stress.

Animals↗

PubMed1998

Arginine vasotocin receptor in the vagina of the oviduct of the hen.

The presence of receptor for arginine vasotocin (AVT) in the vagina of the oviduct of the hen was demonstrated by the use of radioligand binding assays on membrane fractions of the tissue. The binding to [125I]AVT was highly competitive with unlabeled AVT. Scatchard analysis revealed that the binding sites are of a single class. The equilibrium dissociation constant (Kd) was 0.48+/-0.05 nM (x+/-SEM; n = 6) in laying hens holding a hard-shelled egg in the uterus (shell gland) and 1.01+/-0.02 nM (n = 6) in nonlaying hens. The maximum binding capacity (Bmax) was 0.41+/-0.04 pmol/mg protein (n = 6) in laying hens and 0.81+/-0.01 pmol/mg protein (n = 6) in nonlaying hens. The Kd value of the laying hens varied from 0.39 to 1.20 nM during an oviposition cycle, showing an increase just prior to oviposition, and the Bmax value also varied from 0.30 to 0.66 pmol/mg protein, showing a gradual increase after 6 h prior to oviposition. In the nonlaying hen, both values were almost constant during a 24-h day. The changes in the binding affinity and capacity of AVT receptor of the vagina may be related to oviposition in the hen.

Animals↗

PubMed2003

Arginine vasotocin induces bearing down for oviposition in the hen.

An intravenous injection of arginine vasotocin (AVT) at 16 h before the expected oviposition of the initial egg of an egg-laying sequence caused bearing down for oviposition. Oviposition was increased up to 100% by increasing the dose of AVT up to 1 microg, but it decreased when the dose was 10 and 20 microg. Bearing down was observed even when the oviposition was not induced by the injection of a higher dose of AVT. The results of the present experiment suggest that bearing down is not necessarily caused by the entering of egg into the vagina, but instead may be caused by hormonal stimulus of AVT receptors present in vaginal tissue.

Animals↗

PubMed1984

Sleep in the cat: raphe dorsalis and vasotocin.

To determine the relationship between the raphe nuclei and the sleep-inducing pineal peptide arginine vasotocin (AVT), we investigated the hypnogenic effects of intraventricularly administered AVT in adult cats with electrolytic lesions performed either in the raphe dorsalis nucleus (RDN) or the raphe pontis nucleus (RPN). The following results were observed: (a) RDN lesions produced a quantitative decrease in NREM sleep and a transitory increase in REM sleep and narcoleptic-like alterations of REM sleep in the first 2-5 postoperative days. On the eighth postoperative day, normal (prelesion) sleep parameters were restored. (b) RDN lesions prevented AVT from inducing its hypnogenic effects during the first 5 postoperative days, but the AVT hypnogenic effects reappeared when sleep parameters reached prelesion levels. (c) Although the RPN lesions produced a slight but statistically significant REM sleep decrease, this did not prevent AVT from inducing its hypnogenic effects. We conclude that (a) the anatomic site of AVT action within the brain may be related to RDN; (b) at least some of the sleep disturbances observed after RDN lesions (narcolepsy) may be due to damage of the AVT site of action or to modifications of AVT synthesis or release; (c) RDN belongs to a large neural circuit that includes the pineal gland and habenula, a circuit that may play an important role in the sleep-wake cycle by "fine-tuning" sleep mechanisms. We hypothesize that AVT could be the specific inhibitory neuromodulator of this neural circuit.

Animals↗

PubMed1991

Effect of beta-adrenergic stimulation on uterine contraction in response to arginine vasotocin and prostaglandin F2 alpha in the gecko Hoplodactylus maculatus.

The effects of beta-adrenergic stimulation on uterine contractions occurring in response to arginine vasotocin (AVT) and prostaglandin F2 alpha (PGF2 alpha) were compared during late pregnancy in the viviparous gecko Hoplodactylus maculatus. High doses of AVT (150 or 1,500 ng/g body weight) induced birth in vivo, but PGF2 alpha at doses of up to 2,000 ng/g did not induce birth. The effect of AVT (150 ng/g) on birth rate in vivo was not enhanced by pretreatment 20 min beforehand with the beta-adrenoreceptor antagonist dichloroisoproterenol (2 micrograms/g), whereas the effect of PGF2 alpha (200 ng/g) was markedly enhanced: geckos treated with dichloroisoproterenol and then with PGF2 alpha showed rapid birth-related behavior and gave birth. Isolated uteri showed a tonic contraction in response to AVT (100 ng/ml) and to PGF2 alpha (1,000 ng/ml). Pre-exposure of isolated uteri to the beta-adrenoreceptor agonist isoproterenol (1 microgram/ml) caused relaxation; this pre-exposure did not block the tonic contraction occurring in response to AVT, whereas it completely blocked the tonic contraction induced by PGF2 alpha. We conclude that in H. maculatus, beta-adrenergic stimulation inhibits uterine contractions induced by PGF2 alpha but not those induced by AVT. These data are the first to show that beta-adrenergic stimulation inhibits uterotonic responses to PGF2 alpha in a reptile, and they suggest that the cellular mechanisms by which AVT and PGF2 alpha induce contraction may differ in this species. They also provide further evidence for similarities between mammals and reptiles in the effects of beta-adrenergic stimulation on uterine relaxation.

Animals↗

PubMed1997

Effect of the oxytocin antagonist atosiban (1-deamino-2-D-tyr(OET)-4-thr-8-orn-vasotocin/oxytocin) on nocturanl myometrial contractions, maternal cardiovascular function, transplacental passage, and fetal oxygenation in the pregnant baboon during the last third of gestation.

The oxytocin antagonist, atosiban (1-deamino-2-D-tyr(OET)-4-thr-8-orn-vasotocin/oxytocin), was infused i.v. to chronically instrumented pregnant baboons in the last third of pregnancy. Atosiban (6 microg/kg per min) inhibited myometrial electromyographic activity associated with spontaneous myometrial contractions that occurred around the onset of darkness between 134 and 162 days gestation (term 180 days gestation). The effect of atosiban on maternal heart rate was minimal. Maternal blood pressure remained unaltered during atosiban infusion. Fetal carotid arterial PO2 was unchanged during a 2-h infusion of atosiban. Transplacental passage of atosiban from mother to fetus was assessed at cesarean section under halothane anesthesia in four baboons and in two chronically instrumented fetuses in the absence of anesthesia. The maternal:fetal concentration gradient ranged from 9.2 to 22.8. Maternal atosiban clearance rates were 9.2-16.9 ml/kg per min. In conclusion, atosiban was very effective at inhibiting spontaneously occurring nocturnal myometrial contractions during the last third of gestation in the pregnant baboon. Although atosiban crosses the placenta relatively freely, there was no effect on fetal oxygenation.

Animals↗

PubMed1997

Oxytocin antagonist [1-D(CH2)5,Tyr(ME)2,Thr4,Tyr-NH2(9)]ornithine vasotocin inhibits oxytocin-induced prostaglandin F2alpha release in late-pregnant cows.

The affinity and specificity of an antagonist of oxytocin, [1-D(CH2)5,Tyr(ME)2,Thr4,Tyr-NH2(9)]ornithine vasotocin (OTA), to oxytocin receptors (OTR) in bovine gestational endometrium was determined in displacement experiments with oxytocin (OT) and vasopressin (AVP) analogues and compared to myometrial OTR. OTA had the highest affinity in both tissues. The effect of OTA on OT-induced increase in plasma concentration of 13,14-dihydro-15-keto-prostaglandin F2alpha metabolite (PGFM) was studied in 24 late-pregnant cows. Treatments consisted of i.v. saline; OT (50 IU); OTA (1200 microg); and OTA (400, 1200, or 4000 microg) injected i.v. 5 min before OT (50 IU) (n = 4 each). Samples were collected from jugular vein at 15-min intervals for 30 min before and 3 h after the injection of OT. Progesterone was measured in once-daily samples taken for 7 days after the experiment. OT caused a twofold increase in plasma PGFM within about 60 min (p < 0.005), with levels returning to baseline at 150-180 min; OTA (1200 microg) caused a gradual lowering of basal plasma PGFM over 180 min (p < 0.05). The 400-microg or 1200-microg dose of OTA did not alter OT-induced PGFM response, whereas the 4000-microg dose inhibited it almost completely (p < 0.005). Plasma progesterone declined after the experiment in all cows, with no differences among groups. Because OTA inhibits OT-induced release of endometrial prostaglandin F2alpha it may be a good tocolytic agent.

Animals↗