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The parvocellular vasotocin system of Japanese quail: a developmental and adult model for the study of influences of gonadal hormones on sexually differentiated and behaviorally relevant neural circuits.

Vasotocin (VT; the antidiuretic hormone of birds) is synthesized by diencephalic magnocellular neurons projecting to the neurohypophysis. A sexually dimorphic system of VT-immunoreactive (ir) parvocellular elements has been described within the male medial preoptic nucleus (POM) and the nucleus of the stria terminalis, pars medialis (BSTm). VT-ir fibers are present in many diencephalic and extradiencephalic locations, and quantitative morphometric analyses demonstrated their sexually dimorphic distribution in regions involved in the control of different aspects of reproduction. Moreover, systemic or intracerebroventricular injections of VT markedly inhibit the expression of some aspects of male sexual behavior. In adult animals, circulating levels of testosterone (T) have a profound influence on the VT immunoreactivity within BSTm, POM, and lateral septum. Castration markedly decreases the immunoreaction, whereas T-replacement therapy restores a situation similar to the intact birds. We observed no changes in gonadectomized females treated with T. These changes parallel similar changes in male copulatory behavior (not present in castrated male quail, fully expressed in castrated, T-treated males). The restoration by T of the VT immunoreactivity in castrated male quail could be fully mimicked by a treatment with estradiol (E(2)), suggesting that the aromatization of T into E(2) may play a key limiting role in both the activation of male sexual behavior and the induction of VT synthesis. This dimorphism has an organizational nature: administration of E(2) to quail embryos (a treatment that abolishes male sexual behavior) results in a dramatic decrease of the VT immunoreactivity in sexually dimorphic regions. Conversely, the inhibition of E(2) synthesis during embryonic life (a treatment that stimulates the expression of male copulatory behavior in treated females exposed in adulthood to T) results in a malelike distribution of VT immunoreactivity. The VT parvocellular system of the Japanese quail can therefore be considered an accurate marker of the sexual differentiation of brain circuits mediating copulatory behavior and could be a very sensitive indicator of the activity of estrogenlike substances on neural circuits.

Animals↗

Vasotocin and mesotocin stimulate the biosynthesis of neurosteroids in the frog brain.

The neurohypophysial nonapeptides vasopressin (VP) and oxytocin (OT) modulate a broad range of cognitive and social activities. Notably, in amphibians, vasotocin (VT), the ortholog of mammalian VP, plays a crucial role in the control of sexual behaviors. Because several neurosteroids also regulate reproduction-related behaviors, we investigated the possible effect of VT and the OT ortholog mesotocin (MT) in the control of neurosteroid production. Double immunohistochemical labeling of frog brain sections revealed the presence of VT/MT-positive fibers in close proximity of neurons expressing the steroidogenic enzymes 3beta-hydroxysteroid dehydrogenase/delta5-delta4 isomerase (3beta-HSD) and cytochrome P450 17alpha-hydroxylase/c17, 20-lyase (P450(C17)). High concentrations of VT and MT receptor mRNAs were observed in diencephalic nuclei containing the 3beta-HSD and P450(C17) neuronal populations. Exposure of frog hypothalamic explants to graded concentrations of VT or MT produced a dose-dependent increase in the formation of progesterone, 17-hydroxypregnenolone, 17-hydroxyprogesterone, and dehydroepiandrosterone. The stimulatory effect of VT and MT on neurosteroid biosynthesis was mimicked by VP and OT, as well as by a selective V1b receptor agonist, whereas V2 and OT receptor agonists had no effect. VT-induced neurosteroid production was completely suppressed by selective V1a receptor antagonists and was not affected by V2 and OT receptor antagonists. Concurrently, the effect of MT on neurosteroidogenesis was markedly attenuated by selective OT and V1a receptor antagonists but not by a V2 antagonist. The present study provides the first evidence for a regulatory effect of VT and MT on neurosteroid biosynthesis. These data suggest that neurosteroids may mediate some of the behavioral actions of VT and MT.

Animals↗

Arginine vasotocin binding component in the uterus (shell gland) of the chicken.

The plasma membrane fraction of the uterus of the chicken was found to contain a component that shows specific binding to arginine vasotocin (AVT), arginine vasopressin (AVP) and oxytocin (OT). This binding component possesses a higher affinity to AVT than to AVP or OT, and the affinity to AVT was higher in laying hens than in non-laying hens and immature pullets, while the maximum number of binding sites per mg protein was less in the laying hens. Intramuscular injections of either estradiol-17 beta, progesterone or testosterone into the immature pullets for six consecutive days caused an increase in the affinity and number of binding sites. The results suggest that AVT receptors are present in the chicken uterus and that their binding properties are affected by ovarian steroid hormones.

Animals↗

Arginine vasotocin receptor binding in the hen uterus (shell gland) before and after oviposition.

The binding affinity and capacity of arginine vasotocin (AVT) receptor in the hen uterus changed during a period before and after oviposition. Three hours before oviposition, the binding capacity of the AVT receptor increased. An injection of prostaglandin (PG) F2 alpha caused an increase in the AVT receptor Bmax in the uterus, and indomethacin blocked the normal rise in the AVT receptor Bmax and PGF content prior to oviposition. However, just prior to oviposition, the binding affinity increased with a decrease in the binding capacity. A progesterone injection caused an increase in binding affinity and a decrease in binding capacity of the AVT receptor. The specific binding of the progesterone receptor in the uterus increased 2 h before oviposition and remained high until oviposition. Serum AVT levels increased at oviposition. An injection of AVT caused an increase in the affinity of the AVT receptor with a decrease in the capacity. The change in the affinity and capacity of AVT receptor at oviposition may result from the action of progesterone via increased progesterone receptor binding, and the action of AVT on its own receptors.

Animals↗

Evidence for prolactin-releasing and release-inhibiting effects of melatonin, serotonin and arginine vasotocin.

Adult female Wistar rats (in 12 hr light/12 hr dark) were pinealectomized (PX) or sham-operated (SO) either 21 days after ovariectomy or on the 15-17th day of pregnancy. Ovariectomized (OVX) rats were injected with estrogen and progesterone (EP) 48 hr before decapitation. Melatonin, serotonin or arginine vasotocin (AVT; 50, 100 or 200 micrograms) were administered intravenously into OVX-EP rats 9 days after pineal removal. In PX and SO groups, the same study was done 3 days after delivery. Sera and pituitaries were collected 30 min after injection in order to determine prolactin (PRL) levels. Fifty micrograms melatonin significantly suppressed serum PRL levels in PX-OVX-EP rats and PX postpartum rats, but had not significant effect in SO-OVX-EP or PX postpartum rats. After administration of AVT, serum PRL levels markedly rose in PX and SO rats. These results suggest that melatonin may act not only to stimulate but also to inhibit rat PRL secretion and that the stimulatory function would be superior to its inhibitory function when the pineal gland is intact.

Animals↗

Arginine vasotocin binding to isolated branchial cells of the eel: effect of salinity.

Binding of 125I-labelled arginine vasotocin (AVT) was studied in isolated intact gill cells obtained from eels (Anguilla anguilla) adapted to fresh water (FW) or to sea water (SW). Experiments carried out at 20 degrees C showed maximum and stable binding beyond 10 min of incubation. Specific binding, determined by using labelled peptide in the presence or absence of an excess of unlabelled hormone, represented 30-50% of total and was reversible, with a half-time of less than 5 min. Scatchard plot analysis revealed the presence of a single population of saturable, high-affinity sites. Maximum binding capacity (Bmax: fmol AVT/10(6) cells) and dissociation constant (Kd: nM) were respectively 5.16 and 3.21 in FW and 24.25 and 1.05 in SW. Analysis of chloride cell number and size in gills and of binding characteristics of AVT revealed parallel changes with external salinity. These results are taken as evidence for the direct intervention of neurohypophysial peptides on the gill epithelium of teleost fishes.

Animals↗

Expression of the vasotocin gene in the hypothalamus of intact and osmotically stimulated bullfrogs during metamorphosis.

To study the ontogeny of the vasotocin (VT) system and its contribution to anuran metamorphosis, VT mRNA levels were determined by Northern blot analysis in metamorphosing bullfrog tadpoles. Effects of osmotic stimulation on VT mRNA levels were also analyzed in order to follow the development of osmotic responsiveness of VT neurons. The intensity of hybridization signals for VT mRNA gradually increased during prometamorphic development. The increase became marked thereafter until metamorphic climax. Plasma osmolality and hematocrit remained unchanged before metamorphosis, and increased after metamorphic climax, indicating that climactic tadpoles in a semi-terrestrial environment were in a dehydrated condition. These increases correlated well with the increase in VT mRNA level. Immersion of tadpoles in 30% seawater (approximately 350 mOsmol) for 3 days increased plasma osmolality at all stages. No significant changes were observed in the VT mRNA level in response to this treatment during premetamorphic stages. The VT mRNA levels were significantly higher in the treated tadpoles after preclimax stages. Hyperosmotic treatment also increased hematocrit until early metamorphic climax, but did not alter it in tadpoles at late metamorphic climax. These results suggest that the responsiveness of VT-producing neurons to hyperosmotic or hypovolemic stimulation, or both, is established by the time of the metamorphic climax in bullfrog. The marked increase in VT mRNA levels at metamorphic climax stages of intact individuals is probably induced by dehydration. VT-stimulated water absorption and reabsorption in the target organs probably prevented the increase in hematocrit at late metamorphic climax. Thus VT may contribute importantly to osmoregulatory mechanisms in relation to adaptation to a semi-terrestrial habitat through the metamorphosis.

Animals↗

Reversal by arginine-vasotocin of the effects of pinealectomy on the amount of 5-hydroxytryptamine in the hypothalamus and the concentrations of luteinizing hormone and follicle-stimulating hormone in the plasma of immature male rats.

Pinealectomy in immature (25-day-old) male Wistar rats significantly decreased the content of 5-hydroxytryptamine (5-HT) in the hypothalamus and increased concentrations of plasma LH and FSH 3 days after surgery. Extremely small amounts (10(-4) pg) of arginine-vasotocin (AVT) injected into the third ventricle (pineal recess) 5 min after pinealectomy completely prevented the decrease in the hypothalamic content of 5-HT and the increase in the concentration of LH and FSH in the plasma. It is suggested that AVT prevented the increase of plasma levels of LH and FSH after pinealectomy by increasing levels of 5-HT at postsynaptic receptor sites.

Animals↗

Effects of alpha-melanocyte-stimulating hormone and [8-arginine]-vasotocin upon melanogenesis in hair follicle melanocytes in vitro.

alpha-Melanocyte-stimulating hormone (alpha-MSH) has been shown to act directly on the mammalian melanocyte in short-term cultures of hair follicles obtained from the Siberian hamster. Melanogenesis was stimulated through an increase in tyrosinase activity which resulted in an increase in melanin production. The response of hair follicle melanocytes to alpha-MSH occurred only in follicles taken from moulting animals, implying that they show a discontinuous expression of MSH receptors during the hair follicle growth cycle. Synthetic 1-24 ACTH had no effect on melanogenesis regardless of whether the follicles came from moulting or non-moulting animals. The pineal peptide, [8-arginine]-vasotocin (AVT), inhibited melanin production without a concomitant decrease in tyrosinase activity. In this respect AVT resembled melatonin, although AVT showed a potency ratio of less than half on a molar basis. The action of AVT, like that of melatonin, must ultimately be on some post-tyrosinase step in melanin biosynthesis. In these hair follicle melanocytes AVT seems to bind to specific receptors since neither of the closely related peptides, oxytocin and [8-arginine]-vasopressin, displayed any activity in our culture system.

Adrenocorticotropic Hormone↗

Effects of [1-(3-mercaptopropanoic acid)]-vasopressin and -vasotocin on the pro-oestrous progesterone surge and ovulation in immature female rats.

The desamino analogues of arginine-vasotocin (dAVT) and arginine-vasopressin (dAVP), when administered on the afternoon of pro-oestrus to immature female rats primed with pregnant mare serum, blocked the preovulatory progesterone surge and inhibited ovulation. Statistical analysis revealed no difference in the potency of the two compounds. At a dosage of 2.0 micrograms, dAVT and dAVP yielded a 70 and 87% block of ovulation respectively. Simultaneous administration of dAVT and the antipressor compound [1-deaminopenicillamine-4-valine-8-D-arginine]-vasopressin had no statistically significant inhibitory effect when compared with saline-treated controls. These data suggest that the inhibitory effects of dAVT and dAVP on the progesterone surge and subsequent ovulation are mediated through receptor sites similar to those mediating the pressor actions of the parent hormones.

Animals↗

Neurohypophysial peptides in the human fetus: presence in pituitary extracts of immunoreactive arginine-vasotocin.

A specific and sensitive radioimmunoassay was used to demonstrate the presence of arginine-vasotocin (AVT) in human fetal pituitary extracts. The mean pituitary contents of AVT in extracts of 70-101, 102-136 and 137-172 days gestation were 0.32, 0.8 and 8.1 ng/gland respectively. Combined high-pressure liquid chromatography (HPLC) and radioimmunoassay of pooled extracts at these gestational ages confirmed the presence of AVT. Radioimmunoassay of neonatal pituitary extracts (240-280 days) did not conclusively show AVT to be present, as the material assayed did not show parallelism with dilution. However, combined HPLC and radioimmunoassay of pooled extracts at this gestational age did demonstrate the presence of an immunoreactive AVT peak. Radioimmunoassay analysis of this peak indicated that a relatively small amount of AVT (at least 0.5 ng/gland) was present in the human pituitary gland at term.

Arginine Vasopressin↗

Effect of dehydration, haemorrhage and oviposition on serum concentrations of vasotocin, mesotocin and prolactin in the chicken.

Serum concentrations of arginine vasotocin (AVT), mesotocin and prolactin were determined by radioimmunoassay in Rhode Island Red chickens during and after dehydration, haemorrhage and oviposition. During dehydration increased circulating levels of AVT, mesotocin and prolactin were found. As water deprivation proceeded, marked differences were observed. After an initial rise in serum AVT, mesotocin and prolactin levels during mild and moderate dehydration, concentrations of both AVT and prolactin tended to normalize during continued water deprivation, while those of mesotocin remained high throughout the whole dehydration experiment with the highest at the end of the water-deprivation period. Removal of 5 ml blood at intervals of 10 min during six consecutive time-periods did not affect serum osmolality and circulating levels of AVT and prolactin, but slightly increased mesotocin. These results suggest an osmoregulatory role for AVT and prolactin, whereas mesotocin may be involved in volume control. Finally, 1 min after oviposition, control values of 19.5 +/- 3.4 pmol AVT/1 (n = 9) were raised more than sevenfold to 142.9 +/- 12.5 pmol/l (n = 11). Thereafter, a decline occurred with a half-life for AVT of 13 min with raised serum levels up to 31 min after oviposition. In contrast, the serum concentrations of mesotocin and prolactin remained unaffected by oviposition.

Animals↗

Volumetric control of arginine vasotocin and mesotocin release in the frog (Rana ridibunda).

Adult male frogs (Rana ridibunda) were subjected to several volumetric and osmometric stimuli and the influence on circulating concentrations of arginine vasotocin (AVT) and mesotocin was studied by the use of highly specific radioimmunoassays. During progressive blood withdrawal (haemorrhage group) urine flow decreased to zero, whereas no change occurred in the plasma and urine osmolality. Control levels of 34.3 +/- 7.3 pmol AVT/1 gradually increased up to 638.3 +/- 179.1 pmol/1 (P less than 0.001) after a blood loss of up to 50-60% of the blood volume. Plasma mesotocin concentrations also increased from 42.4 +/- 9.2 to 70.8 +/- 12.0 pmol/1 (n = 7). Hypervolaemia, produced by the repeated intravenous injection of isotonic Ringer solution, increased the urine flow and osmolality compared to controls but had no influence on the plasma levels of AVT and mesotocin. Hypernatraemia without volume change profoundly increased the urine osmolality but the urine flow was not affected; the plasma concentrations of AVT and mesotocin remained at the control level. Finally, during a 1-h immobilization stress a pronounced antidiuresis occurred in the presence of a constant plasma and urine osmolality and control plasma levels of AVT and mesotocin. It is concluded that the release of AVT and, to a smaller extent, of mesotocin is under volumetric control.

Animals↗

Modulation of arginine vasotocin secretion in chicks by dehydration during embryogenesis.

The influence of low humidity during incubation (LHI) on arginine vasotocin (AVT) secretion in male and female White Leghorn chicken embryos and chicks was investigated. LHI embryos were incubated from day 8 (E8) to E18 at a relative humidity of 32-35%, while control embryos were incubated at 58-62%. In the control groups, plasma AVT gradually increased from E15 to E18, increased sharply during hatching and returned to the E18 level within 4 h. LHI did not alter the level of AVT until E18. LHI E19 males had concentrations of AVT twice as high as those of control males (39 +/- 3.9 vs 15 +/- 1.6 pmol/l, P < 0.001). Augmentation persisted during hatching and 4 h afterwards. In LHI females additional elevation reached significant levels only at 4 h after hatching (58 +/- 13 vs 16 +/- 1.7 pmol/l, P < 0.01). Deprivation of water caused an increase in plasma sodium concentrations in the 2-day-old chicks of the control groups, while no significant changes were observed in the LHI group. Deprivation of water together with a high environmental temperature caused an additional increase in AVT only in the LHI groups (42 +/- 5.2 vs 22 +/- 3.9 pmol/l, P < 0.01 for males; 32 +/- 3.4 vs 19 +/- 2.8 pmol/l, P < 0.01 for females). The results clearly showed that LHI leads to a more pronounced response of the AVT system on subsequent osmotic and non-osmotic challenges, especially in males. Embryonal activation of the AVT system probably helps the organism to cope with hatching and posthatching stresses.

Animals↗

Diurnal variation in uterine estrogen receptors in immature female rats--inhibition by arginine vasotocin.

A diurnal variation in uterine estrogen receptors from immature female rats was observed with the peak occurring during the mid-light phase (noon) of the cycle. Although no dose response was noted, all concentrations of arginine vasotocin ranging from 5 x 10(-7) to 5 x 10(-9) significantly inhibited binding of 3H-estradiol to estrogen receptors in the cytosol fraction of uteri obtained from immature female rats either during the light or dark phase of the photoperiod.

Aging↗

Effect of vasotocin on plasma free fatty acid level in the migrating anadromous sea lamprey.

The effect of injections of arginine vasotocin (AVT) on plasma free fatty acid (FFA) levels was studied in anadromous sea lampreys collected in the St. John River, New Brunswick, during their upstream spawning migration. Plasma FFA was significantly higher in lampreys injected with a single dose of 1 000 mU vasotocin/kg body weight than in those receiving only the vehicle solution, the difference being the greater at 90 than at 30 min post-injection. The significance of AVT in migration is discussed.

Acclimatization↗

Arginine vasotocin induces free fatty acid release from avian adipose tissue in vitro.

The effect of arginine vasotocin (AVT) on free fatty acid (FFA) release from pigeon adipose tissue slices incubated in vitro was studied. AVT at concentrations of 0.5 microgram/ml and 5.0 micrograms/ml was found to produce significant increases in the release of FFA from the adipose tissue. This augmentation of FFA release did not require the presence of hydrocortisone.

Adipose Tissue↗

Effect of acute and chronic cardiac denervation on osmotic release of vasotocin in chickens.

Hypo- and hyperosmotic NaCl were infused intravenously to examine osmotic release of arginine vasotocin (AVT) in anesthetized, acutely cardiac-denervated chickens and in conscious, chronically denervated birds. Mean arterial blood pressure was consistently higher in denervated compared to sham-operated chickens but heart rates were similar in experimental and control groups. Plasma AVT concentrations (pAVT) were significantly higher than controls in acutely, but not chronically, denervated chickens. The slope of the regression line relating pAVT to plasma sodium concentration was higher in denervated birds indicating that removal of cardiac receptor activity increases the osmotic sensitivity of the AVT system. The results suggest that cardiac end-net receptor activity may participate in the regulation of blood pressure and can modulate the release of antidiuretic hormone in the chicken.

Animals↗