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PubMed1991

The distribution of corticotropin-releasing factor--immunoreactive neurons and nerve fibers in the brain of the snake, Natrix maura. Coexistence with arginine vasotocin and mesotocin.

The anatomical distribution of neurons and nerve fibers containing corticotropin-releasing factor (CRF) has been studied in the brain of the snake, Natrix maura, by means of immunocytochemistry using an antiserum against rat CRF. To test the possible coexistence of CRF with the neurohypophysial peptides arginine vasotocin (AVT) and mesotocin (MST) adjacent sections were stained with antisera against the two latter peptides. CRF-immunoreactive (CRF-IR) neurons exist in the paraventricular nucleus (PVN). In some neurons of the PVN, coexistence of CRF with MST or of CRF with AVT has been shown. Numerous CRF-IR fibers run along the hypothalamo-hypophysial tract and end in the outer layer of the median eminence. In addition, some fibers reach the neural lobe of the hypophysis. CRF-IR perikarya have also been identified in the following locations: dorsal cortex, nucleus accumbens, amygdala, subfornical organ, lamina terminalis, nucleus of the paraventricular organ, nucleus of the oculomotor nerve, nucleus of the trigeminal nerve, and reticular formation. In addition to all these locations CRF-IR fibers were also observed in the lateral septum, supraoptic nucleus, habenula, lateral forebrain bundle, paraventricular organ, hypothalamic ventromedial nucleus, raphe and interpeduncular nuclei.

Animals↗
PubMed1994

Development of vasotocin pathways in the bullfrog brain.

The brain of adult bullfrogs (Rana catesbeiana) contains six populations of cells which are immunoreactive for the neurohypophysial peptide arginine vasotocin (AVT). It is unknown when some of these cell populations first appear during development and when the sexual differences in AVT distribution first become apparent. We therefore used immunocytochemistry to examine development of AVT pathways in developing bullfrog tadpoles and in newly metamorphosed froglets of both sexes. AVT-immunoreactive (AVT-ir) cells were already present in the three diencephalic areas (magnocellular preoptic nucleus, suprachiasmatic nucleus and hypothalamus) at stage III (Taylor and Kollros stages), the earliest stage examined. Cell size in the magnocellular nucleus was not bimodally distributed in either tadpoles or froglets. AVT-ir cells in the telencephalic septal nucleus and amygdala did not appear until stage VI. There was no sexual difference in the density of AVT-ir cells or fibers in the amygdala of tadpoles or froglets. Finally, cells in the hindbrain pretrigeminal nucleus appeared much later--after stage XX. Thus, different populations of neurons begin to express AVT at unique times during development. The sexual dimorphism in AVT content observed in the amygdala of adult bullfrogs must appear during juvenile development or at adulthood.

Animals↗
PubMed1982

Central administration of arginine vasotocin: effects on exploratory behavior in the rat.

Synthetic arginine vasotocin (AVT) was infused into rat brains either by intraventricular administration or by local infusion on the pineal body. Subsequently, exploratory behavior was analyzed in a hole board. The behavioral effects induced by this peptide were dependent on the time of day, i. e. the light or the dark phase. High intraventricular doses (0.4 microgram) administered during the light phase altered exploratory activity such that the number of hole visits was increased, while the duration of each visit was decreased; lower doses produced no effect. In contrast, during the dark phase peripineal infusion of AVT (10(-4) pg) attenuated the number of hole visits and increased the mean duration of the visits. The strongest effects were obtained with peripineal applications during the dark phase. This treatment also resulted in significantly lowered levels of pineal melatonin.

Animals↗
PubMed1980

Radioimmunoassay for arginine-vasotocin (AVT) in serum of Pekin ducks: AVT concentrations after adaptation to fresh water and salt water.

A radioimmunoassay for arginine-vasotocin (AVT), the antidiuretic principle in birds, was developed using the high cross-reactivity of AVT with an AVP antiserum raised in rabbits. This assay is specific for the measurement of AVT in serum of birds. The sensitivity and precision is such that serum AVT concentrations above 0.5 fmol/ml can be measured quantitatively. A serum AVT concentration of 5.1 +/- 1.4 fmol/ml was found in normally hydrated, fresh water adapted ducks with a serum osmolality of 293.7 +/- 2.2 mosmol/kg. When the same animals were acutely hydrated, no or < 0.5 fmol/ml AVT was present at an osmolality of 289.8 +/- 2.4 mosmol/kg. In salt water adapted ducks with a serum osmolality of 333.0 +/- 7.6 mosmol/kg, a serum AVT of 22.7 +/- 3.0 fmol/ml was found. When fresh water adapted ducks were hydrated and infused with different doses of AVT, a linear relationship between serum AVT and the amount infused was obtained. During AVT infusions, serum osmolality and urine flow rate were negatively correlated and urine osmolality was positively correlated with serum AVT.

Adaptation, Physiological↗
PubMed1980

Serum arginine-vasotocin (AVT) and afferent and central control of osmoregulation in conscious Pekin ducks.

In conscious Pekin ducks adapted either to fresh water or to hypertonic saline (1.9%) as drinking fluid, urinary excretion, salt gland secretion and the serum concentration of radioimmunoassayable arginine-vasotocin (AVT) were examined with regard to their afferent and central control. The experiments were carried out under conditions of water diuresis, osmotic diuresis or supraorbital salt gland secretion, which were induced by continuous infusions of appropriate solutions. Temporary bilateral vagus blockade caused rises in AVT serum concentration accompanied by antidiuresis in hydrated ducks and by inhibition of salt gland secretion in salt-stressed ducks. Rostral brainstem cooling caused decreases of AVT serum concentration and water diuresis in ducks under osmotic diuresis and reduction of AVT serum concentration and inhibition of salt gland secretion in salt-stressed ducks. Cerebral osmotic stimulation in hydrated ducks by intracarotid injection or by intracerebroventricular microinfusion of hypertonic NaCl solutions caused antidiuretic reactions associated with rises of AVT serum concentration.

Animals↗
PubMed1984

Blood volume changes and arginine vasotocin (AVT) blood concentration in conscious fresh water and salt water adapted ducks.

Blood volume changes consisting in the removal and reinfusion respectively, of 10% of the estimated blood volume (23.2 ml on average) were induced to determine their effects on the blood concentration of arginine vasotocin (AVT), the antidiuretic hormone (ADH) of birds, in fresh water adapted ducks (water ducks) with blood osmolalities and ADH concentrations similar to those of normally hydrated mammals, and in salt water adapted ducks (salt ducks) with chronically elevated blood osmolalities and ADH concentrations. The investigations were carried out in steady state conditions, when infusion of 1 ml . min-1 of isotonic saline was matched by the excretion in water ducks and when infusion of 0.4 ml . min-1 of 1,000 mosmolal saline was matched by the salt gland excretion in the salt ducks. After blood removal, AVT blood concentration (mean +/- SE) increased from 6.5 +/- 0.4 to 8.4 +/- 0.6 pg . ml-1 in water ducks and from 18.1 +/- 1.6 to 22.6 +/- 1.9 pg . ml-1 in salt ducks. The respective blood osmolalities of 297.4 +/- 1.4 and 318.6 +/- 3.3 mOsm . kg-1 did not change. Reinfusion of the blood after steady-state conditions had been reattained decreased blood AVT from 7.9 +/- 0.7 to 6.7 +/- 0.5 pg . ml-1 in water ducks. In the salt ducks AVT concentration had already returned to the control level before blood reinfusion which induced no further reduction. The blood osmolalities remained unchanged in both groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Adaptation, Physiological↗
PubMed1986

Plasma levels of arginine vasotocin, prolactin, aldosterone and corticosterone during prolonged dehydration in the domestic fowl: effect of dietary NaCl.

Three groups of White Plymouth Rock laying hens were adapted to three levels of dietary NaCl: low-NaCl food with tap water (LOW), high-NaCl food (1% NaCl w/w added) with tap water (HT), and high-NaCl food with 0.5% NaCl for drinking (HS). The birds were subjected to water deprivation (dehydration) for 18 days. Blood sampling was done at 2-4 day intervals. Plasma concentrations of arginine vasotocin (AVT), prolactin (PRL), aldosterone (ALDO) and corticosterone (CS) were determined by radioimmunoassay. Plasma osmolality, sodium, chloride, and potassium were also determined. In the normally hydrated hens fully adapted to the diets, there was a stepwise increase from LOW to HS in plasma osmolality (305, 315, 332 mOsm, for LOW, HT and HS, respectively), [Na+] (144, 153, 161 mM) and [Cl-] (109, 119, 127 mM) as well as in [AVT] (6, 14, 18 pg/ml) and [PRL] (16, 24, 34 ng/ml). Regressing [AVT] on osmolality gave a slope of 0.30 pg . ml-1/mOsm and a threshold of 273 mOsm. The slope of [PRL] on osmolality was 0.73 ng . ml-1/mOsm. The correlation coefficient of [AVT] and [PRL] was 0.67. LOW had high [ALDO] (165 pg/ml) which was suppressed to low levels in HT and HS (5-8 pg/ml), while [CS] was the same in all groups (0.9-1.1 ng/ml). Plasma [K+] was decreased in the high-NaCl groups (5.8 mM in LOW, 4.4 and 4.7 mM in HT and HS). Dehydration resulted within 2 days generally in a sharp (5-15%) increase in osmolality, [Na+] and [Cl-], which thereafter increased more slowly during the remaining 16 days in all groups, with the slowest increase in LOW. The levels of osmolality [Na+] and [Cl-] were 5% lower in LOW than in HT and HS, which showed the same levels during the dehydration period. Plasma [AVT] and [PRL] increased 2-4 fold within 2 days of dehydration; [AVT] reached a plateau at 29 pg/ml in all groups, but [PRL] continued to rise in all groups, fastest in LOW, reaching similar levels in all groups after 14-18 days of dehydration, about 85 ng/ml. The correlation coefficient of [AVT] and [PRL] was decreased by half (to 0.32) during dehydration. Plasma [ALDO] increased in all groups with dehydration, 1.7 fold in LOW and 3-6 fold in HT and HS, but the levels reached in HT and HS were only 15-30% of that seen in LOW.(ABSTRACT TRUNCATED AT 400 WORDS)

Adrenal Cortex Hormones↗
PubMed1986

Osmotic stimuli and NaCl-intake in the fowl; release of arginine vasotocin and prolactin.

White Plymouth Rock hens were fed a high- and a low-NaCl content of the diet. The two groups were exposed to moderate dehydration, to intra-arterial hyperosmotic NaCl-loading, or to injection of physiological doses of arginine vasotocin (AVT). The plasma levels of AVT and prolactin were measured by accurate and sensitive radioimmunoassay and the osmolality and Na, K (and Cl) concentrations also measured for 48 h after dehydration, and for 60-90 min after NaCl-loading or AVT-injection. The plasma concentration of AVT after a given increase of plasma osmolality was in all experiments found higher in the low- as compared to the high-NaCl diet group. The average difference was 0.2 pg/ml X mOsm. The intra-arterial injection of AVT resulted in a strictly mono-exponential fall over the next hour with an average half-life of 6.3 min without any difference between the high- and the low-NaCl diet groups. It is concluded (a) that the release of prolactin after osmotic stimulation is most likely caused by a direct effect of osmolality (or Na concentration) and not by AVT, (b) that the release of AVT is influenced by the NaCl-intake in a direction which tends to maintain extracellular volume.

Animals↗
PubMed1987

Gene conversion in avian mesotocin and vasotocin genes: a recurrent mechanism linking two neurohypophysial precursor lineages?

Amino acid sequences of the first half of MSEL- and VLDV-neurophysins from goose and chicken have been determined. Identical substitutions in positions 17, 18, 35, 36 and 41 of both neurophysins of a given species when compared with their mammalian counterparts suggest a gene conversion between vasotocin--MSEL-neurophysin and mesotocin--VLDV-neurophysin genes. This event, which has already been observed for three mammalian species, seems recurrent and would link the evolution of the two neurohypophysial hormone precursors.

Amino Acid Sequence↗
PubMed1987

Morning administration of melatonin antagonizes the antigonadotrophic effect of afternoon injections of melatonin, 5-methoxytryptophol and arginine vasotocin in intact mice.

In intact prepubertal mice kept under a long photoperiod subcutaneous afternoon (4:30 p.m.) injections (20 micrograms) but not morning (9:30 a.m.) injections (100 micrograms) of melatonin and 5-methoxytryptophol elicited antigonadotrophic effects. A morning dose (100 micrograms) of methoxytryptophol abolished the antigonadotrophic effects of afternoon injections of methoxytryptophol. Similarly the antigonadotrophic effects of afternoon injections of melatonin, 5-methoxytryptophol and arginine vasotocin could be partially destroyed by 100 micrograms and completely abolished by 1 mg melatonin administered in the morning.

Animals↗
PubMed2006

Timing of parturition in two species of viviparous lizard: influences of beta-adrenergic stimulation and temperature upon uterine responses to arginine vasotocin (AVT).

The southern snow skink Niveoscincus microlepidotus is a viviparous alpine lizard with biennial reproduction, in which embryos are fully developed before winter but parturition is delayed until spring. We aimed to determine whether, in this species, in vitro uterine preparations are responsive to arginine vasotocin (AVT) and prostaglandin (PGF(2alpha)) in autumn and spring, and whether pre-treatment with the beta-adrenergic agonist isoproterenol decreases the effectiveness of AVT in stimulating uterine contractions. Using the spotted snow skink (Niveoscincus ocellatus), an annually breeding species, we aimed to determine influences of temperature and the beta-adrenergic system upon the response to AVT in vivo. In both N. microlepidotus and N. ocellatus females are more responsive to AVT than to PGF(2alpha), and that the response to AVT is decreased, but not prevented, by beta-adrenergic stimulation. In N. microlepidotus, uteri are equally responsive in both seasons to the hormones administered. In N. ocellatus environmental conditions, specifically, temperature, modulate the response to AVT in vivo with the time to parturition increasing as temperature decreases. We conclude that in these viviparous squamates the endocrine cascade leading to parturition is modulated by the beta-adrenergic system, and that this may reflect the mechanism by which the timing of parturition is tied to suitable environmental conditions.

Adrenergic beta-Agonists↗
PubMed2004

Nitric oxide inhibits arginine-vasotocin-induced increase of water osmotic permeability in frog urinary bladder.

The present study addressed the question of whether nitric oxide (NO) participates in regulation of osmotic water permeability in the urinary bladder of the frog Rana temporaria L. Experiments were carried out on isolated, paired hemi-bladders filled with amphibian Ringer solution diluted 1:10 with distilled water. Sodium nitroprusside (SNP, 125-250 micro M), an NO donor, markedly attenuated the increase of osmotic water flow elicited by arginine-vasotocin (AVT) (AVT 10(-10) M: 2.20+/-0.26; AVT plus 200 micro M SNP: 1.21+/-0.15 micro l/min cm(2), n=20, P<0.001). This effect of SNP was apparent only in the presence of 50 micro M zaprinast, an inhibitor of the cGMP-specific phosphodiesterase-5 (PDE5). In the presence of zaprinast, SNP elevated cGMP production significantly both in control and AVT-stimulated urinary bladders, but had no effect on the level of cAMP (AVT 5 x 10(-10) M: 7.6+/-0.6; AVT plus SNP 200 micro M: 7.5+/-0.4 pmol/mg protein, n=8, N.S.). 1 H-[1,2,4]-oxadiazole-[4,3-a]-quinoxalin-1-one (ODQ, 25-100 micro M), an inhibitor of soluble guanylate cyclase, enhanced the AVT-induced water flow, decreased the SNP-stimulated increase of cGMP in the bladder tissue and almost abolished the inhibitory effect of SNP on the AVT-induced hydroosmotic response. 8-( p-Chlorophenylthio)-cGMP (8-pCPT-cGMP, 25 or 50 micro M), a membrane-permeable cGMP analogue specific for cGMP-dependent protein kinase (PKG), inhibited, whereas 2 micro M KT-5823, an inhibitor of PKG, significantly stimulated the increase of water flow induced by AVT. The inhibitory effect of SNP on AVT-induced water flow was almost completely reversed by KT-5823, but not by 50-100 micro M erythro-9-[2-hydroxy-3-nonyl]adenine (EHNA), an inhibitor of cGMP-activated PDE2. Immunohistochemistry of urinary bladder slices with antibodies against different types of NO synthase (NOS) revealed a positive immunostaining for neuronal NOS (nNOS) in the mucosal epithelium. These results suggest that in the frog urinary bladder endogenous NO is involved in regulation of water osmotic permeability. NO inhibits the AVT-induced increase of water flow at least partly by activation of PKG, which interferes with the hydroosmotic effect of AVT probably at (a) post-cAMP step(s).

Adenine↗
PubMed2005

Immunocytochemical studies on translocation of phosphorylated aquaporin-h2 protein in granular cells of the frog urinary bladder before and after stimulation with vasotocin.

We have generated a specific antibody against phosphorylated aquaporin-h2 (pAQP-h2) protein to investigate the role of phosphorylation in the translocation of AQP-h2 protein within the granule cells of the urinary bladder of the frog (Hyla japonica). The antibody was generated against a synthetic peptide (ST-160) corresponding to amino acids 255-268, with a phosphorylated Ser-262, a residue that is putatively phosphorylated by protein A kinase. Using this antibody, we found, by Western blot analysis, that phosphorylation of the AQP-h2 protein rapidly increased within 2 min after vasotocin (AVT) stimulation and remained at a higher than normal level for 15 min. Moreover, quantitative immunoelectron microscopy indicated that the location of the AQP-h2 protein dramatically changed after AVT stimulation. Before stimulation, pAQP-h2 protein was localized in only a small number of intracellular vesicles near the nucleus of the granular cells, whereas the labeling density of the intracellular vesicles and the apical membrane rapidly increased after stimulation. This finding was also confirmed by the results of an immunofluorescence study. Thus, phosphorylation of AQP-h2 protein seems to be essential for translocation of the protein from the cytoplasmic pool to the apical plasma membrane of the granular cells in frog urinary bladder.

Amino Acid Sequence↗
PubMed1997

Sexual dimorphism of arg-vasotocin gene expressing neurons in the telencephalon and dorsal diencephalon of the domestic fowl. An immunocytochemical and in situ hybridization study.

A strong sex dimorphism in the distribution of immunoreactive arginine-vasotocin (AVT) and AVT mRNA was observed in telencephalic and dorsal diencephalic areas of the domestic fowl using immunocytochemistry and in situ hybridization. Two subgroups of immunoreactive parvocellular perikarya surrounded by dense plexus of immunoreactive fibres were found within the bed nucleus of the stria terminalis and the dorsal part of the diencephalic paraventricular region of males. No signs of immunoreactivity were observed within corresponding regions of the female brain. Instead, in females a few scattered weakly stained perikarya were observed rostrally to the level of the anterior commissure, juxtapositioned to the nucleus accumbens and the floor of the lateral ventricle. The distribution of AVT mRNA containing cell profiles fully confirmed the immunocytochemical findings. Osmotic stress induced by water deprivation for 48 h had no influence on the number of immunoreactive or AVT mRNA containing parvocellular cell bodies. However, it resulted in an increase of immunoreactive cell area in the bed nucleus of the stria terminalis and dorsal diencephalon of 5. 9 and 11.7%, respectively. We suggest that the sexually dimorphic vasotocinergic circuit may be involved in the co-ordination of behavioural and autonomic functions in response to environmental stress.

Animals↗
PubMed1999

Arginine vasotocin mRNA revealed by in situ hybridization in bovine pineal gland cells.

Arginine vasopressin (AVP) is the main antidiuretic hormone in mammals and arginine vasotocin (AVT) in submammalian vertebrates. The possibility that the genetic material encoding AVT is maintained in mammals is controversial. In this study, we investigated by radioactive in situ hybridization the possible presence of the mRNA encoding AVP and AVT, and using immunocytochemistry the presence of structures immunoreactive for AVP and AVT in the bovine pineal gland. In situ hybridization was performed by use of 35S-labelled oligoprobes. Immunocytochemistry was performed using specific polyclonal rabbit antibodies and the avidin-biotin-complex method. In situ hybridization revealed positive signals for both AVP mRNA and AVT mRNA in a few cells scattered throughout the pineal body. Immunocytochemistry revealed thin AVP-immunoreactive fibres in the pineal stalk and the pineal gland. It also revealed staining of several AVT-immunoreactive nerve fibres in both the pineal stalk and the gland. In addition, polyhedral, neuron-like cell bodies from which two to three processes emerged were also AVT-immunoreactive. Thus, our investigation shows the presence of AVP/AVT-immunoreactive cellular structures in the bovine pineal gland. Our data further show the presence of mRNAs encoding both AVT and AVP. We therefore suggest that AVT mRNA is translated into an AVT-like peptide in the bovine pineal.

Animals↗
PubMed1987

One-step processing of the amphibian vasotocin precursor: structure of a frog (Rana esculenta) "big" neurophysin.

Vasotocin-associated neurophysin (MSEL-neurophysin) from the frog Rana esculenta has been isolated and sequenced through tryptic and staphylococcal proteinase peptides and cyanogen bromide fragments. This protein appears homologous to the mammalian vasopressin-associated neurophysin with a C-terminal glycopeptide extension homologous to the mammalian copeptin. In contrast to the two-step processing of mammalian vasopressin/MSEL-neurophysin/copeptin precursor, a single cleavage is therefore involved in the processing of the amphibian vasotocin/neurophysin precursor. It appears that the physiological release of the vasopressin-like hormone from the N-terminal end of the protein precursor is not dependent upon a previous trimming of the C-terminal copeptin-like moiety.

Amino Acid Sequence↗