Immunoreactivity distribution of vasotocin and vasoactive intestinal peptide in brain nuclei of two songbird species with different breeding systems.

Vasopressin influences social behaviour in mammals, in particular social recognition and bonding. However, much less is known about its avian analogue, vasotocin, although vasotocin appears to modulate singing behaviour and agonistic interactions together with vasoactive intestinal peptide (VIP) in some songbirds. The objectives of our study were to compare the expression of vasotocin and VIP in brain nuclei hypothetised to be part of the social behavioural network, i.e. septal areas, bed nucleus of the stria terminalis and medial preoptic nucleus (POM), in two songbird species in the wild: the blue tit (Cyanistes caeruleus) and European penduline tit (Remiz pendulinus). These two closely related passerine birds differ in their pair bonding and mating systems: blue tits are socially monogamous with extensive pair bond lasting for several months, whereas in the European penduline tit, pair bond is short and it dissolves during or after laying of the eggs. The two species did not differ in the distribution of vasotocin in the observed brain regions; however, VIP was more abundant in all three regions of penduline tits than in blue tits. We found a sex difference in favour of males in the distribution of vasotocin- and VIP-immunoreactive neurones, fibres and terminals in all three regions in penduline tits. In blue tits, such gender differences were only observed in the POM. The limited differences between the two species suggest that the levels of vasotocin and VIP in the socially relevant brain regions are likely influenced by many other social or environmental factors than just by differences in the duration of pair bonding.

Brain arginine vasotocin immunoreactivity differs between urban and desert curve-billed thrashers, Toxostoma curvirostre: relationships with territoriality and stress physiology.

The neuropeptide arginine vasotocin (AVT: the avian homolog of vasopressin) has numerous functional roles including mediating social behaviors, coregulating the adrenocortical stress response and maintaining water balance. These functions of AVT make it susceptible to environmental influence, yet little is understood concerning the variation in the AVT system across habitats. In this study, AVT immunoreactivity was compared between male curve-billed thrashers, Toxostoma curvirostre, from native Sonoran Desert locations and those within the city of Phoenix, Ariz. Previous research found that urban thrashers are more responsive to territorial intrusion, secrete more corticosterone (CORT) during capture stress, and they may also have greater access to water than desert counterparts. Variation in AVT immunoreactivity was also related to levels of plasma CORT and osmolality, and with behavioral responses to a simulated territorial intrusion. Birds from these two habitats showed different AVT immunoreactive patterns in two brain regions: the paraventricular nucleus of the hypothalamus and the medial bed nucleus of the stria terminalis (BSTM), a part of the limbic system. Immunoreactive AVT within the paraventricular nucleus was associated with plasma CORT levels in urban, but not desert, birds, but no such association with osmolality was observed in birds from either habitat. The total number of BSTM AVT-immunoreactive cells was related to a decreased responsiveness to territorial intrusion. These data suggest that divergence in the AVT system between urban and desert thrashers may help explain observed differences in both the adrenocortical stress response and territorial behavior between populations. Whether differences in water availability between habitats contribute to population differences in the brain AVT system is unknown.

Sexual dimorphism in the vasotocin system of the bullfrog (Rana catesbeiana).

Arginine vasotocin (AVT) is widespread in amphibian brains, where its levels have been correlated with reproductive behaviors. To better understand which neural systems are involved in central actions of AVT, we used immunocytochemistry to compare the distribution of AVT in the brains of male and female bullfrogs (Rana catesbeiana). AVT-immunoreactive cells were observed in the septal nucleus, amygdala pars lateralis, magnocellular preoptic area, suprachiasmatic nucleus, and hypothalamus. AVT-immunoreactive cells were also found in the pretrigeminal nucleus, but only in animals killed in the fall. Immunoreactive fibers were broadly distributed in hypothalamic and extrahypothalamic areas. The most obvious sex differences were found in the amygdala pars lateralis, where the density of immunoreactive cells and fibers was significantly greater in male than in female bullfrogs. In addition, in the habenular nucleus, males had a denser distribution of AVT-immunoreactive fibers than females. In the suprachiasmatic nucleus, AVT-immunoreactive cells were larger in females than in males but did not differ in number. Since the areas that showed sex differences in AVT distribution have also been implicated in control of reproductive behaviors, they may form the neural substrates for the effects of AVT on sexually dimorphic behaviors in amphibians.

Organization of vasotocin-immunoreactive cells and fibers in the canary brain.

The distribution of vasotocin (VT)-immunoreactive neuronal perikarya and fibers in the canary (Serinus canaria) was investigated with immunohistological techniques. The results suggest that most VT-stained cell bodies are located in three diencephalic regions. First, a large number of densely packed neurons are found in the paraventricular nucleus (PVN) and the anterior preoptic nucleus. Neurons here vary widely in size and shape. Small-size rounded neurons and large-size multipolar neurons appear to concentrate in separate subdivisions. Second, a series of loosely organized cell groups of medium- to large-size cells occurs in the lateral parts of the hypothalamus. These aggregates of neurons apparently correspond to subdivisions of the supraoptic nucleus (SON). Third, diffusely distributed, lightly stained cells are found dorsal to the paraventricular nucleus in the dorsal diencephalon. A number of cells of this group seem to be located in the basal septal area and bed nucleus of the stria terminalis. Immunoreactive fibers and varicosities concentrate in brain regions that are associated with neuroendocrine, autonomic, and limbic functions. Axons from the PVN and SON form compact bundles of the hypothalamohypophysial tract in the lateral hypothalamus and then funnel into the internal zone of the medium eminence (ME). Furthermore, a heavy innervation seems to be present in the palisadal, external zone of the ME. A substantial number of fibers appear to leave the PVN toward extrahypothalamic areas. Most extrahypothalamic VT fibers innervate telencephalic and brainstem regions that are thought to be involved in mediation of limbic and autonomic functions. These areas include the lateral and medial septum, the lateral habenula, the substantia grisea centralis, the area ventralis (Tsai), the locus coeruleus, raphe nuclei, the nucleus tractus solitarii, and lateral medulla. In addition, fibers with immunoreactivity for VT innervate structures such as the optic tectum and the nucleus ovoidalis that have been implicated in sensory processing of visual and auditory information. Finally, VT fibers and varicosities occur in centers including the nucleus robustus archistriatalis and nucleus intercollicularis that have been implicated in vocal control.

Distribution of immunoreactive mesotocin and vasotocin in the brain and pituitary of chickens.

The distribution of vasotocin and mesotocin in the pituitary and central nervous system in male chickens was determined using radioimmunoassays. Neither peptide was detected in the pineal. Mesotocin, but not vasotocin, was detected in the cerebellum. Both peptides were found in the septal area, archistriatum, paleostriatum, optic lobe, anterior, medial and posterior hypothalamus, midbrain, pons, medulla oblongata, and the anterior and posterior pituitary. Equal amounts of the 2 peptides were present in the septal area, archistriatum and anterior hypothalamus whereas vasotocin was more abundant (2- to 10-fold) in the paleostriatum, optic lobe, midbrain, and pituitary. The amount of mesotocin was about twice that of vasotocin in the medulla oblongata and the medial and posterior hypothalamus. The wide distribution of vasotocin and mesotocin in extrahypothalamic sites in the central nervous system suggests that the peptides may, as in mammals, have a role in a variety of autonomic and endocrine regulatory processes in chickens.

Radioimmunoassay of arginine vasopressin, oxytocin and arginine vasotocin-like material in the human pineal gland.

Specific radioimmunoassays (RIAs) have been developed for arginine-vasopressin (AVP), oxytocin (OT), and arginine-vasotocin (AVT): they can detect 2.5 pg, 5 pg and 1 pg of hormone respectively. The three antisera employed in the RIAs in the present study all had a specificity that discriminated between the three neuropeptides and thus allowed their accurate measurement in each of 157 human pineal glands. Immunoreactive AVP was found to have a mean concentration of 306.4/+-27.6 (SE) pg/gland in the 1389 pineals where it was at level higher than the detection limit. Immunoreactive OT was determined as 386.3/+-42.1 pg/gland in the 110 pineals where it was measurable, and the mean apparent immunoreactive AVT content was 44.6/+-3.6 pg/gland in the 110 pineals where it was found at detectable levels. Cross-reactivity with AVP or OT cannot account for the immunoreactive peptide content of human adult pineal glands was demonstrated as a function of: (1) sex, (2) time of death, (3) cause of death, (4) age (18 to 85 yr old) or (5) delay between death and gland removal over the range of 4 to 48 hr. While the presence of AVP and OT is not surprising, indications of an immunoreactive AVT may in fact reflect a peptide which is closely related to AVT and cross-reacts in the AVT RIA.

Arginine vasotocin induces sexual behavior of newts by acting on cells in the brain.

To investigate whether arginine vasotocin (AVT) acts on target cells in the brain of Taricha granulosa (a urodele amphibian), the behavioral effects of intracerebroventricular (ICV) and intraperitoneal (IP) injections of AVT were compared. Male newts exhibited the greatest sexual activity (amplectic clasping) following an ICV injection of 0.1 microgram AVT. Another study showed that nanogram quantities of AVT, administered ICV, stimulated the behavior. An ICV injection of an antagonist to arginine vasopressin, d(CH2)5Tyr(Me)AVP, or an anti-AVT immune serum significantly inhibited the sexual behavior. Intracranial implants of 17 beta-estradiol (E2) or 5 alpha-dihydrotestosterone (DHT) in castrated males maintained the behavioral response to an injection of AVT. Another study found that an IP injection of DHT or E2 did not increase the incidence of newt sexual behavior during the 8 hours following the injection.

Immunoreactive vasotocin in the zebra finch brain (Taeniopygia guttata).

UNLABELLED: The distribution of vasotocin (VT)-immunoreactive cells and fibers in the zebra finch brain was studied with immunocytochemical techniques. A large number of VT-immunoreactive cells was found in the nucleus preopticus anterior and nucleus paraventricularis. In the lateral parts of the hypothalamus diffusely organized VT-immunoreactive cells were present. Some of these cells were found close to and embedded in the optic tract. An extra-hypothalamic parvocellular cell group was found at the level of the anterior commissure in the dorsal diencephalon. VT-immunoreactive fibers and varicosities were present in a number of extrahypothalamic regions i.e. the septum, the area ventralis of Tsai, the lateral habenula, the optic tectum, the substantia grisea centralis, the nucleus tractus solitarii, the lateral medulla, the nucleus intercollicularis and in the archistriatum surrounding the nucleus robustus archistriatalis. The VT-immunostaining in the lateral septum and dorsal diencephalon showed no sex differences and can not be influenced by testosterone administration. IN CONCLUSION: the topography of the central VT-immunoreactive network is similar to that in another song bird i.e. the canary; but seasonal and testosterone-dependent changes in VT-immunostaining in the lateral septum and dorsal diencephalon are only observed in the canary.

Narcoleptic-like alterations of the sleep cycle in cats induced by a specific vasotocin antiserum.

A specific arginine vasotocin (AVT) antiserum (10 microliter) injected undiluted into the lateral ventricle of unanesthetized cats, induced sleep onset REM periods, markedly increased the number of REM periods, and dramatically decreased REM sleep latency. The amount of total REM sleep significantly increased and the amount of total NREM sleep significantly decreased. Neither a specific arginine vasopressin or oxytocin antiserum nor a normal rabbit serum injected undiluted into the lateral ventricle in the same volume (10 microliter) was able to reproduce the effects of the AVT antiserum. It is concluded that pineal AVT is physiologically involved in the induction and/or circadian organization of REM sleep.

Vasotocin increases quiet sleep and suppresses active sleep in newborn cats. Opposite effects after vasotocin immunoneutralization.

One picogram of synthetic arginine vasotocin (AVT) administered into the lateral ventricle of newborn cats aged 7-9 days, increased the amount of quiet sleep and markedly decreased the amount of active sleep. Ten pg AVT completely suppressed active sleep during the recording time of 3 hr. The increase in the quiet sleep occurred within 5-7 min and lasted for the entire recording time of 3 hr. An undiluted specific AVT antiserum (10 microliters) administered into the lateral ventricle, induced opposite effects suppressing quiet sleep and increasing active sleep. It is concluded that in newborn cats both AVT and the AVT antiserum induce the same hypnogenic effects as in the adult cats.

Effect of testosterone on the distribution of vasotocin immunoreactivity in the brain of the zebra finch, Taeniopygia guttata castanotis.

The distribution of vasopressin or vasotocin immunoreactive cells and fibers in the lateral septum and the bed nucleus of the stria terminalis are sexually dimorphic in many vertebrates including several species of birds examined to date. We examined the vasotocin-like immunoreactivity in the zebra finch brain. Male birds had a higher level of immunoreactive staining in some telencephalic and diencephalic regions. The density of immunostaining increased in the testosterone-treated females to levels typically seen in males. The sexual dimorphism and testosterone dependence of the vasotocin-like immunoreactivity are similar to that found in the canary. Thus this pattern of vasotocin localization and testosterone dependence may be a general feature in brains of passerine songbirds.

Distribution of immunoreactive vasotocin and mesotocin in the chicken gastrointestinal tract.

Immunoreactive arginine vasotocin (AVT) and mesotocin (MT) were measured in heart, breast muscle, adrenals, testes, and different parts of the gastrointestinal tract in adult male chickens. Neither of the peptides were detected in liver, testis, heart and breast muscle. The amounts of AVT and MT in the adrenals were 167 +/- 25 and 669 +/- 198 pg/gland, respectively. Considerable amounts of immunoreactive peptides were found in the gastrointestinal tract with the highest concentration in the proventriculus (4.18 +/- 0.31 ng AVT and 16.58 +/- 0.86 ng MT per organ). Dose-response curves of duodenal and proventriculus extracts were parallel with synthetic AVT and MT standards.

Distribution of nonapeptide systems in the forebrain of an African cichlid fish, Astatotilapia burtoni.

Nonapeptides and their receptors have important functions in mediating social behavior across vertebrates. Where these nonapeptides are synthesized in the brain has been studied extensively in most vertebrate lineages, yet we know relatively little about the neural distribution of nonapeptide receptors outside of mammals. As nonapeptides play influential roles in behavioral regulation in all vertebrates, including teleost fish, we mapped the distributions of the receptors for arginine vasotocin (AVT; homolog of arginine vasopressin) and isotocin (IST; homolog of oxytocin/mesotocin) throughout the forebrain of Astatotilapia burtoni, an African cichlid fish with behavioral phenotypes that are plastic and reversible based on the immediate social environment. We characterized the distribution of the AVT V1a2 receptor (V1aR) and the IST receptor (ITR) using both immunohistochemistry for protein detection and in situ hybridization for mRNA detection, as well as AVT and IST using immunohistochemistry. Expression of the neuropeptide receptors was widely distributed throughout the fore- and midbrain, including the proposed teleost homologs of the mammalian amygdala complex, striatum, hypothalamus, and ventral tegmental area. We conclude that although the location of nonapeptide synthesis is restricted compared to tetrapod vertebrates, the distribution of nonapeptide receptors is highly conserved across taxa. Our results significantly extend our knowledge of where nonapeptides act in the brains of teleosts to mediate social transitions and behavior.

Arginine vasotocin and glomerular filtration rate in saltwater-acclimated ducks.

We examined the relationship between whole kidney glomerular filtration rate (GRF) and the plasma concentration of immunoreactive arginine vasotocin (AVT), the avian antidiuretic hormone, in saltwater-acclimated ducks. During steady-state diuresis, driven by infusion of sodium chloride solutions, transient reductions of [14C]inulin clearance (3 ml X min-1 X kg-1) occurred when plasma AVT concentrations were roughly doubled by systemic injection of synthetic AVT or after stimulation of endogenous AVT release by perfusion of the third ventricle with hypertonic artificial cerebrospinal fluid (CSF). Transient increases in GFR occurred when plasma AVT was reduced during inhibition of its endogenous release by hypotonic ventricle perfusion. GFR also increased after injection of AVT antiserum but returned to control values within 30 min, while plasma AVT concentration remained very low for at least 1 day. During antidiuresis evoked by infusion of strongly hypertonic saline, GFR values estimated from plasma disappearance curves of [125I]iothalamate were not different from the GFR values estimated subsequently with the same method in the same ducks made diuretic by hypotonic saline infusions, although AVT concentrations were depressed during the latter as compared with the former infusion. Factors other than AVT must be important for the control of GFR during sustained osmotic stress.

Arginine vasotocin immunoreactivity in hypothalamic and extrahypothalamic areas of an amphibian brain.

The distribution of arginine vasotocin immunoreactivity (irAVT) was investigated in the brain of male rough-skinned newts (Amphibia: Taricha granulosa) using microdissection and radioimmunoassay techniques. The highest levels of irAVT were observed in the neuroendocrine hypothalamo-hypophysial system. However, detectable quantities of irAVT (greater than 0.40 ng/mg protein) also were observed in extrahypothalamic areas such as pallium, amygdala, striatum and thalamic, tectal and brainstem nuclei. No irAVT was detected in the olfactory bulb, rostral telencephalon, or in specific areas of the caudal telencephalon and diencephalon. These results indicate that, in amphibians, AVT is distributed among a wide range of brain areas and that there is regional specificity in irAVT concentrations.

An antiserum that recognizes mesotocin and isotocin: development of a homologous radioimmunoassay for plasma mesotocin in chickens (Gallus domesticus).

This report presents characteristics of an antiserum raised in a rabbit immunized with synthetic mesotocin (MT) conjugated to bovine thyroglobulin. Cross-reactivity studies indicate that the antiserum (Kl-II) recognizes the carboxyl-terminal "tail" of MT and isotocin (IT). A homologous, disequilibrium radioimmunoassay (RIA) for MT has been developed that can detect less than 1 pg of peptide. Plasma was extracted with octadecasilyl-silica. Recovery of MT from plasma was correlated with the amount added and averaged 70%. Different volumes of plasma and posterior pituitary extract, when measured in the assay system, yielded inhibition curves that were parallel with standard MT. Immunoreactive MT and AVT of plasma and neural lobe coeluted with synthetic standards after gel filtration. The ED50 of a heterologous, sequential saturation RIA for IT was 17.4 pg, suggesting that the MT antiserum may be useful for measuring the oxytocin-like principle in bony fishes. Immunoreactive MT in plasma of cockerels increased and decreased with iv infusion of hypo- and hyperosmotic saline, respectively. The changes in plasma MT were inversely related to osmolality. Hyperosmotic saline infusion resulted in correlated increases in plasma AVT and osmolality. The data suggest that MT may be released by dilution and/or expansion of extracellular fluid in chickens.

Opposite effects of vasotocin and of a specific vasotocin antiserum on active sleep of kittens.

The effects of intracerebroventricularly administered synthetic arginine vasotocin (AVT) or undiluted AVT, vasopressin (AVP) and oxytocin (OT) antisera on active sleep (AS) of newborn kittens have been investigated in comparison with rabbit serum control. In contrast to AVP and OT antisera, AVT antiserum has produced opposite effects on AS as AVT itself. Since after 10 microliter of undiluted AVT antiserum the percentage of AS did not decrease under 20% and even after 100 microliter AS did not decrease under 5%, it is concluded that, at least during perinatal life, AVT could be considered as a neuromodulator with AS-promoting effect.

Characterisation of multiple immunoreactive neurons in the central nervous system of the pond snail Lymnaea stagnalis with different fixatives and antisera adsorbed with the homologous and the heterologous antigens.

In the central nervous system of the pond snail Lymnaea stagnalis a large number of elements (cells and fibers) can be identified with antisera (a-FM) to the molluscan cardioactive tetrapeptide FMRFamide (Phe-Met-Arg-Phe-NH2). Of these elements some are also reactive to antivasotocin (a-VT) and/or anti-gastrin (a-Gas). These observations suggest that the a-FM positive elements belong to more than one type. Previous results had already indicated that the immunoreactivity of many a-FM positive cells is influenced by the type of fixation. Taking into account the effects of three fixatives on the reactivity of the cells, and their staining characteristics with the two other antisera used, 8 a-FM positive types could be distinguished. Homologous and heterologous adsorptions were carried out to test the specificity of a-FM, a-VT and a-Gas. After homologous adsorptions no staining was obtained. After heterologous adsorptions only part of the multiple staining cells were identified. This indicates that in a-FM, a-VT and a-Gas in addition to (more) selective IgG molecules, less specific IgG molecules occur that can bind to other peptides than those used to raise the antisera (cross-reaction). The (more) selective IgG molecules in a-FM bind to 6 of the a-FM positive types, suggesting that in L. stagnalis a family of FMRFamide-like substances occurs. This conclusion is sustained by results obtained with a-FM adsorbed with fragments of FMRFamide.(ABSTRACT TRUNCATED AT 250 WORDS)

Extrahypothalamic distribution of vasotocin-immunoreactive fibers and perikarya in the avian central nervous system.

Immunohistochemical analysis of the extrahypothalamic distribution of vasotocin-like immunoreactive elements within the central nervous system of the domestic fowl and Japanese quail, revealed several mesencephalic, pontine and bulbar target areas topographically identifiable. Extrahypothalamic immunopositive perikarya were observed in diencephalic and mesencephalic locations after glutaraldehyde fixation.