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Arginine-vasopressin (AVP) is believed to be positively correlated with the brain edema formation, but the underlying mechanism is still unclear. In this study, we explored the role of the V2 receptors antagonist tolvaptan on brain edema following intracerebral hemorrhage (ICH) with a rat model. Animals were randomly given tolvaptan or vehicle through oral gavage at 12 h, 36 h, and 60 h after ICH surgery. Brain swelling (%), brain water content(BWC), neurological scores, Evans blue fluorescence and blood-brain barrier (BBB) tight junction proteins were measured to evaluate the effect of tolvaptan in ICH. We found that tolvaptan alleviated the brain swelling (%), decreased the BWC growth, and attenuated the neurological deficits after ICH (p 0.05, vs vehicle). What's more, tolvaptan increased the expression of ZO-1 and occludin (p 0.05, vs vehicle), which might be attributed to the down-regulated effects of tolvaptan on MMP-9. These results provided evidence supporting the use of tolvaptan for ICH-induced brain edema. Large animal experiments are required to further explore the efficacy and mechanism of tolvaptan in ICH treatment.

Ischemic stroke is associated with cardiac myocyte vulnerability through some unknown mechanisms. Arginine vasopressin (AVP) may exert considerable function in the relationship of brain damage and heart failure. Danhong injection (DHI) can protect both stroke and heart failure patients with good efficacy in clinics. The aim of this study is to investigate the mechanism of DHI in heart and brain co-protection effects to determine whether AVP plays key role in this course. In the present study, we found that both the supernatant from oxygen-glucose deprivation (OGD) and reperfused primary rat neuronal cells (PRNCs) and AVP treatment caused significant reduction in cell viability and mitochondrial activity in primary rat cardiac myocytes (RCMs). Besides, DHI had the same protective effects with conivaptan, a dual vasopressin V1A and V2 receptor antagonist, in reducing the RCM damage induced by overdose AVP. DHI significantly decreased the injury of both PRNCs and RCMs. Meanwhile, the AVP level was elevated dramatically in OGD and reperfusion PRNCs, and DHI was able to decrease the AVP expression in the injured PRNCs. Therefore, our present results suggested that OGD and reperfusion PRNCs might induce myocyte injury by elevating the AVP expression in PRNCs. The ability of DHI to reinstate AVP level may be one of the mechanisms of its brain and heart co-protection effects.

The possible involvement of oxytocin (OT) in the generation of seizures has not received a lot of attention in the past, although generalized epileptic convulsions were observed in humans following intravenous OT infusion. We here aimed to investigate the effect of exogenous OT administration on seizure susceptibility in C57Bl/6 mice subjected to the pentylenetetrazol (PTZ) model. In addition, we studied via which receptor possible effects on seizure thresholds could be mediated since OT binds to both the OT receptor (OTR) and the vasopressin 1a receptor (V1aR). We showed that C57Bl/6 mice treated with 0.5 mg/kg OT had decreased PTZ thresholds for ear twitch, myoclonic twitch, tail twitch, forelimb clonus and falling. This pronconvulsive effect was reversed by the OTR antagonist L-368.899, however, it was not mimicked by the OTR agonist carbetocin (CBT). Nevertheless, CBT had antidepressant-like effects in the forced swim test that could be reversed by L-368.899. These experiments shed some doubt on the involvement of OTR in the observed effect of OT on seizure thresholds. Therefore, we investigated the role of the V1aR as a possible mediator of the proconvulsive effects of OT. We found that the proconvulsive effects of both arginine vasopressin and OT were reversed by the V1aR antagonist SR49059. In summary, OT has proconvulsive effects in our mouse model of generalized seizures that could not be mimicked by CBT. Our results suggest that the binding of OT to V1aRs is the most plausible explanation for the proconvulsive effects of OT.

Our previous study has proven that hypothalamic paraventricular nucleus (PVN) played a role in the antinociception. The central bioactive substances involving in the PVN regulating antinociception were investigated in the rat. The results showed that electrical stimulation of the PVN increased the pain threshold, and L-glutamate sodium injection into the PVN elevated the pain threshold, but the PVN cauterization decreased the pain threshold; pain stimulation raised the arginine vasopressin (AVP), not oxytocin (OXT), leucine-enkephalin (L-Ek), beta-endorphin (beta-Ep) and DynorphinA1-13 (DynA1-13) concentrations in the PVN tissue using micropunch method, heightened AVP, L-Ek, beta-Ep and DynA1-13, not OXT concentrations in the PVN perfuse liquid, and reduced the number of AVP-, not OXT, L-Ek, beta-Ep and DynA1-13-immunoreactive neurons in the PVN especially in the posterior magnocellular part of the PVN using immunocytochemistry. There was a negative relationship between the PVN AVP concentration and the pain threshold; pain stimulation enhanced the AVP, not OXT mRNA expression in the PVN using in situ hybridization and RT-PCR; intraventricular injection of anti-AVP serum completely reversed L-glutamate sodium injection into the PVN-induced antinociception, and administration of naloxone - the opiate peptide antagonist, partly blocked this L-glutamate sodium effect, but anti-OXT serum pretreatment did not influence this L-glutamate sodium effect; L-glutamate sodium injection into the PVN-induced analgesia was inhibited by V2 receptor antagonist - d(CH2)5[D-Ile2, Ile4, Ala-NH2(9)]AVP, not V1 receptor antagonist - d(CH2)5Tyr(Me)AVP. The data suggested that the PVN was limited to the central AVP, not OXT, which was through V2, not V1 receptors influencing the endogenous opiate peptide system, to regulate antinociception.

MicroRNAs are a class of small RNAs that regulate the expression of target mRNAs by inhibiting translation or destabilizing target mRNAs. miR-126 is a microRNA that is highly enriched in endothelial cells. miR-126 has been found to promote angiogenesis and inhibit vascular inflammation in endothelial cells by repressing three target genes Sprouty-related EVH1 domain-containing protein 1 (SPRED1), phosphoinositol-3 kinase regulatory subunit 2 (PIK3R2), and vascular cell adhesion molecule 1 (VCAM1). Our previous study showed that the expression of miR-126 was downregulated after spinal cord injury (SCI). Therefore, we wanted to examine whether upregulation of miR-126 could promote angiogenesis, inhibit inflammation, and exert a positive effect on recovery after contusion SCI. In this study, we found that increased levels of miR-126 promoted angiogenesis, and inhibited leukocyte extravasation into the injured spinal cord, which was concurrent with downregulation of mRNA and protein expression of three validated miR-126 target genes, SPRED1, PIK3R2, and VCAM1. Moreover, a dose-dependent effect of miR-126 was observed in rescuing tissue damage and improving the functional deficit after SCI. Thus, the present study indicated that miR-126 played an important role in angiogenesis and inflammation after SCI.

Earlier autoradiographic studies from our laboratory detected vasopressin recognition sites in the mammalian cerebral cortex [R.E. Brinton, K.W. Gee, J.K. Wamsley, T.P. Davis, H.I. Yamamura, Regional distribution of putative vasopressin receptors in rat brain and pituitary by quantitative autoradiography, Proc. Natl. Acad. Sci. U. S.A., 81 (1984) 7248-7252; C. Chen, R.D. Brinton, T.J. Shors, R.F. Thompson, Vasopressin induction of long-lasting potentiation of synaptic transmission in the dentate gyrus, Hippocampus, 3 (1993) 193-204]. More recently, we have detected mRNA for the V1a vasopressin receptors (V1aRs) in cultured cortical neurons [R.S. Yamazaki, Q. Chen, S.S. Schreiber, R.D. Brinton, V1a Vasopressin receptor mRNA expression in cultured neurons, astroglia, and oligodendroglia of rat cerebral cortex, Mol. Brain Res., 45 (1996) 138-140]. To determine whether these recognition sites are functional receptors, we have pursued the signal transduction mechanism associated with the V1a vasopressin receptor in enriched cultures of cortical neurons. Results of these studies demonstrate that exposure of cortical neurons to the selective V1 vasopressin receptor agonist, [Phe2,Orn8]-vasotocin, (V1 agonist) induced a significant accumulation of [3H]inositol-1-phosphate ([3H]IP1). V1 agonist-induced accumulation of [3H]IP1 was concentration dependent and exhibited a linear dose response curve. Time course analysis of V1 agonist-induced accumulation of [3H]IP1 revealed a significant increase by 20 min which then decreased gradually over the remaining 60 min observation period. V1 agonist-induced accumulation of [3H]IP1 was blocked by a selective V1a vasopressin receptor antagonist, (Phenylac1, D-Tyr(Me)2, Arg6,8, Lys-NH29)-vasopressin. Results of calcium fluorometry studies indicated that V1 agonist exposure induced a marked and sustained rise in intracellular calcium which was abolished in the absence of extracellular calcium. The loss of the rise in intracellular calcium was not due to a failure to induce PIP2 hydrolysis since activation of the phosphatidylinositol pathway occurred in the absence of extracellular calcium. V1 agonist activation of calcium influx was then investigated. V1 agonist-induced 45Ca2+ uptake was concentration dependent with a biphasic time course at 250 nM. Preincubation with the L-type calcium channel blocker, nifedipine, blocked V1 agonist-induced calcium influx suggesting V1 agonist-induced L-type calcium channel activation in cortical neurons. Furthermore, V1 agonist-induced calcium influx was blocked by both bisindolyleimide I (PKC inhibitor) and U-73122 (PLC inhibitor) suggesting a modulation of V1 agonist-induced L-type calcium channel activation by downstream components of the phosphatidylinositol signaling pathway such as protein kinase C. These results indicate that in cultured cortical neurons, V1a vasopressin receptor activation leads to induction of the phosphatidylinositol signaling pathway, influx of extracellular calcium via L-type calcium channel activation, and a rise in intracellular calcium which is dependent on V1a receptor activated influx of extracellular calcium. These data are the first to demonstrate an effector mechanism for the V1 vasopressin receptor in the cerebral cortex and provide a potential biochemical mechanism that may underlie vasopressin enhancement of memory function.

Studies were performed on the central antidiuretic actions via the tachykinin NK-3 receptor in the rat hypothalamic paraventricular nucleus (PVN). Microinjections of the selective tachykinin NK-3 receptor agonist senktide (2-200 pmol) into the PVN resulted in prolonged inhibition of urine output in water-loaded rats, its effect being dose-dependent. The antidiuretic action of senktide was blocked by pretreatment with the vasopressin V2 receptor antagonist OPC-31260 (1 mg/kg, i.v.), but not by microinjection of the angiotensin II AT-1 receptor antagonist losartan (1 nmol) into the PVN. NK-3 receptor mRNA was strongly detected in the magnocellular part of the PVN and the supraoptic nucleus (SON) of the hypothalamus as detected by in situ hybridization histochemistry. Moreover, [3H]senktide binding sites were also detected in the PVN and the SON by receptor autoradiography. These findings suggest that NK-3 receptors in the PVN may be involved in water regulation by stimulation of vasopressin secretion from the posterior pituitary gland, and that vasopressin caused water reabsorbtion via the kidney V2 receptor.

Increased plasma osmolality results in increased central as well as peripheral release of vasopressin. Experiments were carried out to determine whether, in this circumstance, vasopressin can act centrally to modulate its peripheral release. Prior to the start of a thirty-min i.v. infusion of 2.5 M or 0.15 M NaCl, the rats were given an intracerebroventricular (i.c.v.) injection of a peptide V 1 /V 2 vasopressin antagonist (2 μg), OPC-31260 (60 μg), a non-peptide V 2 antagonist, or 1-desamino-8- d -arginine vasopressin (dDAVP, 5 ng), a V 2 agonist. Experiments with the peptide antagonist were carried out in male and non-estrous female rats. Since there were no differences between males and females in the measured responses, experiments with the other two drugs were carried out only in males. Pretreatment with either the V 1 /V 2 antagonist or the V 2 antagonist enhanced the increase in plasma vasopressin levels in response to the hypertonic saline infusion by about 50% at the end of 30 min. dDAVP, on the other hand, had no effect. None of the i.c.v. drugs had an affect on either the pressor or bradycardic responses to hypertonic saline infusion. These observations suggest that vasopressin can act centrally in a negative feedback fashion to attenuate its own release into the peripheral circulation in response to increased plasma osmolality.

Vasopressin (AVP) receptor binding within hypothalamic sites was compared between cycling and lactating female golden hamsters. The density of AVP receptor binding was analyzed by quantitative autoradiography within the ventrolateral hypothalamus and dorsomedial hypothalamic nucleus. Lactation was correlated with a disappearance of AVP receptor binding within the ventrolateral hypothalamus. In contrast, lactation was associated with a two- to three-fold increase in the density of AVP receptor binding within the dorsomedial hypothalamic nucleus. These results suggest that AVP receptor binding within the ventrolateral hypothalamus is responsive to gonadal hormones in female golden hamsters. However, the increase in binding observed within the dorsomedial hypothalamus may be related to other neurobiological changes associated with lactation.

Arginine-vasopressin (AVP) is a nonapeptide of hypothalamic origin that has been shown to exert many important cognitive and physiological functions in neurons and terminals of both the central and peripheral nervous system (CNS and PNS). Here we report for the first time that AVP induced an increase in intracellular Ca²⁺ concentration ([Ca²⁺](i)) in non-neuronal cells isolated from the rat dorsal root ganglion (DRG) and cultured in vitro. The ratiometric [Ca²⁺](i) measurements showed that AVP evoked [Ca²⁺](i) responses in the non-neuronal cells and these concentration-dependent (100 pM to 1 μM) responses increased with days in vitro in culture, reaching a maximum amplitude after 4-5 day. Immunostaining by anti-S-100 antibody revealed that more than 70% of S-100 positive cells were AVP-responsive, indicating that glial cells responded to AVP and increased their [Ca²⁺](i). The responses were inhibited by depletion of the intracellular Ca²⁺ stores or in the presence of inhibitors of phospholipase C, indicating a metabotropic response involving inositol trisphosphate, and were mediated by the V₁ subclass of AVP receptors, as evidenced by the use of the specific blockers for V₁ and OT receptors, (d(CH₂)₅¹,Tyr(Me)²,Arg⁸)-Vasopressin and (d(CH₂)₅¹,Tyr(Me)²,Thr⁴,Orn⁸,des-Gly-NH₂⁹)-Vasotocin, respectively. V(1a) but not V(1b) receptor mRNA was expressed sustainably through the culture period in cultured DRG cells. These results suggest that AVP modulates the activity of DRG glial cells via activation of V(1a) receptor.

We recently demonstrated that the neural peptide vasopressin (AVP) can act as a neurotrophic factor for hippocampal nerve cells in culture. Because the neurotrophic effect of vasopressin is mediated by the V1 receptor [11], we investigated AVP activation of calcium signaling pathways in cultured hippocampal neurons. Results of this investigation demonstrate that exposure of cultured hippocampal neurons prelabeled with [3H]myo-inositol to vasopressin induced a significant accumulation of [3H]inositol-1-phosphate ([3H]IP1). The selective V1 vasopressin receptor agonist, [Phe2, Orn2]vasotocin, induced a significant accumulation of [3H]IP1 whereas a selective V2 vasopressin receptor agonist, [deamino1, D-Arg8]-vasopressin, did not. Moreover, V1 agonist-induced accumulation of [3H]IP1 was blocked by the selective V1 vasopressin receptor antagonist d(CH2)5[Tyr(Me)2]-vasopressin. V1 agonist-induced accumulation of [3H]IP1 was concentration dependent and exhibited a steep inverted U-shaped curve that included both stimulation and inhibition of [3H]IP1 accumulation. Time course analysis of V1 agonist-induced accumulation of [3H]IP1 revealed significant increase by 20 min which continued to be significantly elevated for 60 min. Investigation of the effect of closely related peptides on [3H]IP1 accumulation indicated that the vasopressin metabolite peptide AVP4-9 and oxytocin significantly increased [3H]IP1 accumulation whereas the vasopressin metabolite peptide AVP4-8 did not. AVP4-9 and oxytocin induced [3H]IP1 accumulation were blocked by the V1 vasopressin receptor antagonist d(CH2)5[Tyr(Me)2]-vasopressin. V1 receptor activation was associated with a pronounced rise in intracellular calcium. Results of calcium fluorometry studies indicated that V1 agonist exposure induced a marked and sustained rise in intracellular calcium that exhibited oscillations.(ABSTRACT TRUNCATED AT 250 WORDS)

Intracellular recordings were made from sympathetic preganglionic neurones (SPNs) in transverse slices of thoraco-lumbar spinal cord of young rats (12-20 days old). A small group of SPNs generally having higher membrane potentials (-70 mV) compared to a remaining group (-66 mV) showed spontaneous oscillations of their membrane potential. Oxytocin superfused in concentrations of 0.1-30 microM had four effects on SPNs, inducing slow depolarisation, EPSPs, IPSPs and brief rhythmic oscillations. The slow depolarisation was unaffected by TTX whereas this abolished the other changes. The oxytocin-induced depolarisation was associated with a slow inward current and was not reversed at membrane potentials negative to EK, it increased at more positive potentials and was still present in low Ca2+ and high Mg2+ solutions. These features of the oxytocin induced current are similar to those of the TTX resistant voltage dependent Na+ current described in brainstem autonomic neurones. Vasopressin superfused at concentrations of 0.1 microM to 30 microM had similar effects on SPNs to those of oxytocin. A comparison of the effects of oxytocin and vasopressin on the same neurones revealed that oxytocin was almost 10 times less potent than vasopressin. The effects of oxytocin were not mimicked by a selective oxytocin agonist but were mimicked by a selective vasopressin V1a agonist and blocked by a selective V1a antagonist. Therefore it is concluded that the effects of oxytocin on SPNs are mediated by the vasopressin V1a receptor. It is suggested that oxytocin and vasopressin terminals in the lateral horn are part of a descending system controlling oscillating networks of SPNs in the spinal cord.

Vasopressin binding sites were detected in the brain of the Siberian hamster, using [ 3 H]vasopressin and a 125 I-labelled linear vasopressin antagonist specific for V 1 vasopressin receptors. In the ventromedial and remammillary nuclei, the density of the binding was lower in the females than in the males. The effect of castration and of testosterone replacement was assessed in males. Two distinct effects were observed. Orchidectomy diminished significantly the vasopressin binding in the ventromedial nucleus, an effect which was prevented by implantation of a mini-pump releasing testosterone. On the contrary, in the premammillary nucleus no significant differences were noticed following castration and testosterone treatment. In addition, vasopressin immunoreactivity was examined in males, in females and in castrated males. No sex differences were evident. However, in the bed nucleus of the stria terminalis and the lateral septal nucleus, castration decreased vasopressin immunoreactivity in either sex. This effect of castration was prevented by testosterone. Vasopressin immunoreactivity was detected neither in the ventromedial nor in the premammillary hypothalamic nuclei. Our observations suggest that, in adult Siberian hamster premammillary nucleus, the expression of vasopressin receptors is not controlled by gonadal steroids but is sex related and could be induced during fetal or early postnatal life.

The binding characteristics and central distribution of 125I-Linear AVP antagonist, a new ligand for vasopressin binding sites, are described in the following studies. Saturation studies performed on rat brain septal membranes demonstrated that 125I-Linear AVP antagonist binds to a single class of sites with high affinity (55 pM) and limited capacity (88 fmol/mg protein). In autoradiographic studies, 125I-Linear AVP antagonist labeled brain areas known to contain vasopressin receptors without binding to neurophysins. 125I-Linear AVP antagonist also labeled sites in cortex, hypothalamus, ventral tegmental area and substantia nigra. In competition studies, 125I-Linear AVP antagonist binding was most readily blocked by AVP and a selective V1a agonist. Oxytocin and a selective V2 ligand were effective only in micromolar concentrations. A selective oxytocin agonist was virtually ineffective in blocking 125I-Linear AVP antagonist binding. In regions that contain a high density of oxytocin binding sites, however, oxytocin-displaceable binding was observed. In agreement with studies on peripheral tissues, the binding profile generated from these studies indicates that 125I-Linear AVP antagonist binds to vasopressin receptors of the V1a subtype. These results suggest that 125I-Linear AVP antagonist is a valuable ligand for the study of central AVP receptors.

The cardiovascular activity of neurohypophyseal peptides has been investigated in concious trout bearing intracerebroventricular guide cannula and a permanent intraarterial catheter. Changes in diastolic pressure and heart rate were monitored during the 25-min period following intracerebroventricular or intraarterial administration of arginine vasotocin and related neuropeptides, including arginine vasopressin, (0.62–5 pmol) induced a dose-dependent increase of diastolic and systolic pressures. The onset of the response occured within 3–5 min after intracerebroventricular administration of arginine vasotocin and the maximal increase was reached at 10–15 min. Central administration of vasopressin and oxytocin induced a significant rise in diastolic and systolic pressures at a dose of 5 pmol while hydrin-2 only caused a significant elevation of blood pressure at a dose of 50 pmol. Central administration of mesotocin, isotocin and conopressin-S (5–500 pmol each) had no significant effect on blood pressure. No changes in heart rate occurred after intracerebroventricular injection of any of the seven neuropeptides tested. Intraarterial injection of arginine vasotocin (50 pmol) induced a significant rise in blood pressure and bradycardia. Peripheral injection of the other neuropeptides did not cause any modification of the cardiovascular activity, whatever the doses administered (5–500 pmol). The V1A receptor antagonist [d(CH 2 ) 5 , Tyr(OMe) 2 ]arginine vasopressin had no intrinsic effect on blood pressure and heart rate when injected centrally (50 pmol) or in the peripheral circulation (200 pmol). At the same doses, [d(CH 2 ) 5 , Tyr(OMe) 2 ]arginine vasopressin reduced by 50 and 66%, respectively, the increase in blood pressure evoked by intracerebroventricular (5 pmol) or intraarterial (50 pmol) injections of arginine vasotocin. These data indicate that arginine vasotocin acts centrally to increase blood pressure in consious trout through activation of receptors related to the mammalian V1 receptor type.

An extremely close association exists between the membranes of the neurosecretory endings and the resident astrocytes (pituicytes) of the neurohypophysis. Indeed, synaptoid contacts involving neurosecretory vesicle-containing axons contacting pituicytes have been observed. suggesting pituicytes as targets of the products released from neurosecretory axons. We have investigated the effects of various neural lobe peptides on pituicytes in primary culture from adult neurohypophyses. Using Fura-2 loaded cells and dynamic ratio imaging, we have determined that arginine vasopressin (AVP) or V1- but not V2-receptor agonists, mobilise potuicyte intracellular Ca2+([Ca2+]i) in the absence of extracellular Ca2+. AVP was consistently effective at concentrations of 10 nM or higher in elevating [Ca2+]i by 200–1000 nM. These responses could be blocked by V1-antagonists and were shown to be associated with accumulation of phosphoinositides. Oxytocin was also found to mobilise [ [Ca2+]i but was effective only at higher concentrations than for AVP. Oxytocin-evoked [Ca2+]i elevations were also blocked by V1-antagonists. Raising [K+]0 was ineffective in changing [Ca2+]i suggesting that these cells lack voltage-gated Ca2+ channels. We conclude that pituicytes possess V1-receptors, activation of which mobilises [Ca2+]i, possibly functioning to initiate a Ca2+-activated K+ conductance which could contribute to further depolarisation of secretory terminals and facilitate exocytosis.

Concomitant intracerebroventricular (i.c.v.) injection of anti-arginine vasopressin (AVP) antiserum dose-dependently suppressed the development of analgesic tolerance to daily morphine, 10 mg/kg, s.c., in mice. This suppressive effect of the antiserum was reduced by incubating the antiserum with AVP in vitro, before i.c.v. injection, suggesting that the antiserum inactivates brain AVP to result in the suppression of the development of tolerance in vivo. Similar to the antiserum, both AVP V1 and V2 antagonists given i.c.v., 10 ng and 20 ng/mouse, respectively, suppressed the development of morphine tolerance. Meanwhile, the administration of antiserum dose-dependently recovered morphine analgesia in morphine-tolerant mice and a complete recovery of analgesia was observed at the highest dose of antiserum following the second injection, and the effect of antiserum was maintained for 3 days after its withdrawal. Likewise, 10-100 ng/mouse of AVP V1 receptor antagonist given i.c.v. recovered morphine analgesia partially but significantly in a dose-dependent manner; however, AVP V2 receptor antagonist at the same doses partially recovered analgesic effect but the effect was neither significant nor dose-dependent. These findings suggest that the tolerance developed to morphine can be reversible when disturbing the function of brain AVP, but in addition to the different mechanisms of antiserum, V1 and V2 receptor antagonists, the V1 receptor-mediated mechanism may be more closely concerned in this phenomenon.

Sites which bind oxytocin and vasopressin with high affinity were detected in the brain and upper spinal cord of 12 human subjects, using in vitro light microscopic autoradiography. Tissue sections were incubated with tritiated vasopressin, tritiated oxytocin or an iodinated oxytocin antagonist. The ligand specificity of binding was assessed with unlabelled vasopressin or oxytocin in excess, as well as in competition experiments using synthetic structural analogues. The distribution of vasopressin binding sites differed markedly from that of oxytocin binding sites in the forebrain, while there was overlap in the brainstem. Vasopressin binding sites were detected in the dorsal part of the lateral septal nucleus, in midline nuclei and adjacent intralaminar nuclei of the thalamus, in the hilus of the dentate gyrus, the dorsolateral part of the basal amygdaloid nucleus and the brainstem. The distribution of oxytocin binding sites in the brainstem has been recently reported (Loup et al., 1989). Oxytocin binding sites were also observed in the basal nucleus of Meynert, the nucleus of the vertical limb of the diagonal band of Broca, the ventral part of the lateral septal nucleus, the preoptic/anterior hypothalamic area, the posterior hypothalamic area, and variably in the globus pallidus and ventral pallidum. The presence of oxytocin and vasopressin binding sites in limbic and autonomic areas suggests a neurotransmitter or neuromodulator role for these peptides in the human central nervous system. They may also affect cholinergic transmission in the basal forebrain and consequently play a role in Alzheimer's disease.

The binding site concentration (Bmax) and equilibrium dissociation constant (Kd) for [3H]-arginine vasopressin (AVP) binding sites were measured in limbic brain areas (septum, dorsal hippocampus, amygdala) and liver and kidney of control and osmotically stimulated male Wistar rats. Membrane binding was performed in these five areas 30, 60 and 180 min following intraperitoneal injection of hypertonic saline. This paradigm resulted in no significant change in binding characteristics in the septum, dorsal hippocampus, amygdala and liver from control treated rats. In contrast, the kidney Bmax was significantly reduced 60min following osmotic stimulation, with no effect on affinity. These results also suggest that AVP receptors in the CNS are relatively resistant to regulatory effects of an acute AVP exposure.

Putative receptors for arginine vasotocin (AVT) in the brain of the newt (Taricha granulosa) were measured using quantitative autoradiography with tritium-labelled vasopressin. Specific binding sites were observed in the olfactory bulb, medial (hippocampal) pallium, dorsal pallium, amygdala pars lateralis and tegmental region of the medulla oblongata. In both male and female newts, concentrations of binding sites in the amygdala, but not in the other four areas, were significantly lower in gonadectomized animals than in sham-operated controls. The equilibrium dissociation constants (Kd s ) were not altered by gonadectomy. Since long-term castration abolishes the effect of AVT injection on sexual behaviors, these results support the hypothesis that gonadal steroids maintain sexual behaviors in this amphibian by maintaining AVT receptors in the amygdala.

The distribution and the amount of [3H]oxytocin binding were studied in the brain of adult rats of either sex, as well as in male and female castrates, some of which received injections of estradiol or testosterone. Intact males were treated with an aromatase inhibitor. Castration and inhibition of aromatase activity reduced, whereas estradiol and testosterone increased oxytocin binding, particularly in regions of the brain assumed to be involved in reproductive functions, such as the ventrolateral part of the hypothalamic ventromedial nucleus and the islands of Calleja and neighbouring cell groups. Binding of oxytocin to the uterus was also estrogen-dependent. In the same animals, we also studied the distribution of [3H]vasopressin binding sites present in the brain. It was similar in males and females, and was not affected by experimentally manipulating gonadal hormone levels. In immunocytochemical studies we noticed, as others had previously, that the vasopressin content of certain areas of the rat brain was affected by castration, whereas the oxytocin innervation was not. These results are discussed in relation to the possible functions of oxytocin in the brain and of the lack of correspondence between the immunocytochemical and the autoradiographic data.

In urodele amphibians, the hypothalamic neuropeptide arginine vasotocin and the adrenal steroid corticosterone interact to regulate reproductive behavior by actions in the brain. The present study investigated signal transduction pathways underlying acute effects of vasotocin and corticosterone, presumably mediated via "non-genomic" steroid action, in an amphibian brain. We used Western blot to examine the effects of corticosterone and the vasotocin receptor agonist arginine vasopressin, alone and in combination, on the subcellular localization and proteolytic processing of protein kinase C-alpha (PKCalpha) in tiger salamander brain tissue. Treatment of whole brain minces with vasopressin or vasotocin led to increases in PKCalpha in membrane fractions and concurrent decreases in PKCalpha in cytosolic fractions. Vasopressin or vasotocin treatment also induced the appearance in membrane and cytosolic fractions of a PKCalpha-immunoreactive band that corresponds to PKMalpha, the proteolytically generated, free catalytic subunit of PKCalpha. Treatment with corticosterone alone had no consistent effect on either PKCalpha or PKMalpha in either fraction. However, pretreatment with corticosterone reliably blocked vasopressin-induced increases in cytosolic PKMalpha. These data provide new information about the cellular mechanisms of action of vasopressin and corticosterone in the vertebrate brain and suggest a cellular mechanism by which the two hormones interact to regulate neuronal physiology and behavior.

The present study investigated the hypothesis that social isolation increases aggression by increasing the number of V1a vasopressin receptors in the anterior hypothalamus (AH). Male hamsters were randomly assigned to a group that was allowed to interact with a small nonaggressive hamster three times each week for 3 weeks (socially experienced) or to a group that did not interact socially with other hamsters (social isolates). On the final day of the experiment, hamsters in both groups were placed in a neutral arena with a small, nonaggressive intruder, and agonistic behavior was scored for 10 min. In social isolates, the duration of aggression and the number of attacks were significantly greater than in socially experienced hamsters. There were no significant between-group differences in the latency to the onset of aggression, the number of flank marks or in the duration of defensive/submissive, social or nonsocial behavior. The amount of V1a receptor binding was significantly greater in the AH, the paraventricular nucleus of the hypothalamus and the lateral hypothalamus in the social isolates than in the socially experienced hamsters. The amount of V1a receptor binding was significantly greater in the central amygdala of socially experienced hamsters than in socially isolated hamsters. Serum concentrations of testosterone were significantly higher in the socially experienced hamsters than in social isolates. These data support the hypothesis that social isolation increases aggression by increasing the number of V1a vasopressin receptors in the AH.

We used a phytoestrogen (PE) and a phytoestrogen-free (PE-Free) diet to determine whether or not diet can have neurobehavioral effects on intermale aggression in Syrian hamsters (Mesocricetus auratus). In Experiment 1, 20 adult male hamsters were pre-tested for aggression and then placed on a PE (n=10) or a PE-Free diet (n=10) for 4 weeks in isolation. During week 5, experimental hamsters were exposed to a group-housed, nonaggressive opponent (NAO) for 5 min in a neutral cage arena. PE-fed hamsters exhibited more attacks (33.4+/-6.1) toward the NAO compared to the PE-Free-fed hamsters (18.1+/-4) (p0.05). Interestingly, testosterone in the blood serum was higher in the PE-fed group (11.01+/-1.48 ng/ml) compared to the PE-Free group (6.5+/-0.87 ng/ml). In Experiment 2, 16 juvenile hamsters were weaned onto a PE (n=8) or a PE-Free diet (n=8). After 7 weeks on the diet, experimental hamsters were exposed to a NAO for 5 min in a neutral cage arena. Although the PE group exhibited higher levels of aggressive behavior, there were no statistically significant differences between groups. However, the PE group had higher levels of testosterone (9.0+/-0.95 ng/ml) compared to the PE-Free group (4.6+/-0.98 ng/ml) (p0.05). In addition, analysis of the brains from both experiments revealed differences in binding for vasopressin 1A (V1A) receptors. Optical densities were converted to disintegrating units per min/mg. The PE-Free group had higher levels of V1A receptor binding (2689.93+/-254.8 dpm/mg) compared to the PE group (1907.32+/-136.3 dpm/mg) in the lateral septum (p0.05). In addition, there were differences in the lateral hypothalamus, but the PE group had higher receptor binding (2550.9+/-63.59 dpm/mg) when compared to the PE-Free group (2011.9+/-174.14 dpm/mg) (p0.05). In sum, these data present the first evidence that phytoestrogens can affect aggressive behavior and, concurrently, alter hormonal status and stimulate changes in the brain of male hamsters.

Arginine vasopressin (AVP) is a peptide involved in social behaviors in rodents. To investigate the mechanism underlying the deficits in social behavior induced by blockade of N-methyl- d -aspartate (NMDA) receptors, this study examined the effect of noncompetitive NMDA antagonists on AVP receptor binding and social interaction in the rat. Subchronic phencyclidine (PCP) administration (2 mg/kg/day, 14 days, i.p.) significantly reduced the density of V1a receptor binding sites, labeled by an [125I]-Linear AVP antagonist, in several brain regions. Subchronic treatment with PCP or MK-801 (0.13 mg/kg/day, 14 days, i.p.) impaired social interactions in rats, as has been previously reported. These results suggest that NMDA antagonists have modulatory effects on the central vasopressinergic system and social interaction.

The neurochemical mechanisms involved in post-ictal antinociception remain to be elucidated. Application of electroconvulsive shock (ECS) to rats results in post-ictal antinociception. The objective of this study was to identify endogenous substances that could participate in antinociception during post-ictal depression induced by ECS (70 mA, 60 Hz, 1 s). Antinociception was measured by the rat paw-pressure test, in which increased sensitivity is induced by intraplantar injection of carrageenan. This test proved to be efficient in detecting the electroshock-induced antinociception. Intense post-ictal antinociception was observed over a period of 30 min after the end of the seizure. It was used nonspecific opioid and specific vasopressin antagonists and the prolactin (PRL) release inhibitor to test the reversal of antinociception. Administration of naloxone (5, 7.5 and 10 mg/kg) blocked the post-ictal antinociception. The V(1) (125 microg/kg) and V(2) (250 microg/kg) vasopressin receptor antagonists ([beta-mercapto-beta,beta-cyclopentamethylenepropionyl(1),O-Et-Tyr(2),Val(4),Arg(8)]-vasopressin and [adamantaneacetyl(1),O-Et-d-Tyr(2),Val(4),Abu(6),Arg(8,9)]-vasopressin) also inhibited the nociceptive response. The antinociception blockade was more intense after administration of the V(1) receptor antagonist. Bromocriptine (4, 8 and 12 mg/kg) was able to reverse antinociception behavior during the post-ictal period. Morphine (1, 2 and 4 mg/kg), vasopressin (12.5, 100 and 400 microg/kg) and prolactin (100, 200 and 400 microg/kg) administration promoted a higher nociceptive threshold. It was administered the three substances with their respective antagonists to verify the opioidergic pathway and vasopressin and prolactin release interactions, and as a positive control. We observed that the tested mediators were released in an independent manner, indicating no interference in which other.

We sought to determine whether vasopressin V 1a receptor (V 1a R) mRNA detected in cortical astrocytes [Brain Res. Mol. Brain Res. 45 (1997) 138] was translated into functional receptors by investigating the effector calcium signaling cascade associated with the vasopressin V 1a receptor subtype. Analysis of intracellular calcium dynamics using the calcium-sensitive dye fura-2 AM indicated that exposure of cortical astrocytes to V 1 vasopressin receptor agonist, [Phe 2 ,Orn 8 ]-oxytocin, induced a marked dose-dependent increase in intracellular calcium which was abolished by depletion of extracellular calcium. V 1 agonist treatment induced a rapid increase in calcium signal in both the cytoplasm and nucleus, which was followed by an accumulation of the calcium signal in the nucleus, suggesting translocation of cytoplasmic calcium into the nucleus. The nuclear calcium signal was sustained for several seconds followed by translocation back to the cytoplasm. Following the nuclear-to-cytoplasmic calcium translocation, total free intracellular calcium concentration decreased. The dynamic calcium cytoplasmic and nuclear localization was confirmed by laser scanning confocal microscopy coupled with the calcium-sensitive dye fluo-3 AM. To determine the source of calcium, V 1 agonist-induced 45 Ca 2+ uptake and [ 3 H]IP 1 accumulation were investigated. V 1 agonist induced significant and rapid uptake of 45 Ca 2+ and a significant dose-dependent increase in [ 3 H]IP 1 accumulation in cortical astrocytes. To our knowledge, this is the first documentation of a vasopressin receptor-induced calcium signaling cascade in cortical astrocytes and the first documentation of vasopressin receptor induction of nuclear calcium signaling.

Neurones in the paraventricular nucleus of the hypothalamus project to rostral ventrolateral medullary spinally projecting vasomotor neurones. We studied the excitatory action and the role of glutamate and vasopressin in this pathway in anaesthetised rats. A five barrel micropipette assembly was used for extracellular recording of neuronal activity and for microiontophoresis of drugs into the vicinity of identified medullary vasomotor neurones. Iontophoresis of l -glutamate or vasopressin into the vicinity of a vasomotor neurone increased activity, effects which were blocked by simultaneous iontophoretic application of a glutamate receptor antagonist, or a vasopressin V1a antagonist respectively. Paraventricular neurones were activated either by microinjecting d , l -homocysteic acid or by disinhibition by microinjecting bicuculline. The excitatory effects on vasomotor neurones, of paraventricular nucleus stimulation at some sites were prevented by simultaneous microiontophoretic application of kynurenic acid or at other sites by application of V1a antagonist. Neither antagonist altered the ongoing activity of the vasomotor neurones. Therefore, glutamate or vasopressin may act as excitatory neurotransmitters at synapses of paraventricular neurones on rostral ventrolateral medullary vasomotor neurones.

In the present study, we have examined neurochemical correlates that may be involved in the differential cardiovascular responses observed in normotensive and hypertensive rats during stress. Using a restraint stress paradigm, both normotensive Wistar Kyoto (WKY) and Spontaneously Hypertensive rats (SHR) underwent acute (1 h restraint in a perspex tube), chronic (1 h restraint for ten consecutive days) or no restraint (control) stress. Following cessation of restraint, rats were processed by incubating sections of brain stem and kidney with [125I]-HO-LVA (0.03 nM) or [125I]Sar(1)Ile(8)-AngiotensinII (0.5 nM), in the presence of PD123319 (10 microM) or losartan (10 microM), to determine the distribution and density of vasopressin V(1A), angiotensin AT(1) and AT(2) receptors, respectively. Analysis of autoradiograms indicated changes in the density of radioligand binding in acutely and chronically-stressed rats, as compared to controls. For example, V(1A) binding in the medial nucleus tractus solitarius (SolM) decreased in the WKY but increased in the SHR. AT(1) binding in SolM did not significantly change in the WKY but decreased in the SHR with repeated restraint. In kidney slices, AT(1) binding decreased with stress in the WKY (-17%) but increased in SHR (+10-15%). AT(2) binding in the kidney showed a pattern similar to that of AT(1) binding in SHR, but not WKY. Graded increases in V(1A) binding were measured in kidney medulla and cortex of both strains (+50-60% with chronic restraint). These results suggest that physiological adaptation to restraint is associated with specific changes in V(1A), AT(1) and AT(2) receptor density within brain nuclei and kidney.

Accurel devices were used to administer AVP or AVP-antagonist into the lateral ventricle of the rat brain for 7 days. Neither AVP nor antagonist altered total binding site concentration in either septum or amygdala. However, treatment with antagonist caused a marked decrease in the affinity of the receptor for agonist in both the septum and amygdala.

The distribution of vasopressin AVP producing cells, their projections and AVP receptors was examined in the brain of common . marmosets Callithrix jacchus using in situ hybridization, immunocytochemistry and receptor autoradiography. Clusters of cells labeled . . . for AVP mRNA or stained for AVP immunoreactivity AVP-ir were found in the paraventricular PVN , supraoptic SON and . suprachiasmatic nuclei SCN of the hypothalamus. Scattered AVP producing cells were also found in the lateral hypothalamus and the . bed nucleus of the stria terminalis BST . Neither AVP mRNA-labeled nor AVP-ir cells were detected in the amygdala. Although AVP-ir fibers were evident outside of the hypothalamic-neurohypophyseal tract, a plexus of fibers in the lateral septum, as observed in the rat brain, was not detected. Receptor autoradiography using 125 I-linear-AVP revealed specific binding for AVP receptors in the nucleus accumbens, diagonal band, lateral septum, the BST, SCN, PVN, amygdala, anterodorsal and ventromedial nucleus of the hypothalamus, indicating sites for central AVP action in the marmoset brain. Together, these data provide a comprehensive picture of AVP pathways in the marmoset brain, demonstrating differences from rodents in the distribution of cell bodies, fibers and receptors. q 1997 Elsevier Science B.V.

We have investigated the distribution of vasopressin binding sites in the brain of male and female adult mice using a radio-iodinated ligand and film autoradiography. Vasopressin receptors were uncovered in various regions of the brain including the basal nucleus of Meynert, the substantia innominata, the hypothalamic paraventricular nucleus, the substantia nigra pars compacta and the hypoglossal nucleus. A sex-related difference in the expression of vasopressin receptors was seen in the medial preoptic area/anterior hypothalamus corresponding to the rat sexually dimorphic nucleus in the rat and in the hypothalamic mammillary nuclei. In both structures the autoradiographic labeling is more intense in females than in males. These observations confirm that vasopressin binding sites are present in the hypothalamic preoptic area of most species examined so far and that sex-related expression of neuropeptide receptors could trigger sex-related behavioral differences.

Vasopressin binding sites were determined in the choroid plexus of five Alzheimer's disease patients and five non-demented controls using the125I-labelled linear V1a-antagonist. The Alzheimer's disease patients showed a twofold increase in the density of vasopressin binding sites, whereas the increase in the affinity constant Kd did not reach significance.

A recently developed ligand with very high affinity and selectivity for the vasopressin (AVP) V1a receptor subtype (i.e. [125I]Linear AVP antagonist ([125I]-LinAntag) was used to describe the distribution of AVP binding sites in the hamster brain, and to determine whether AVP receptor binding was influenced by testicular hormones in sites involved in the regulation of steroid-dependent social behaviors. These studies demonstrated [125I]LinAntag binding in regions of the hamster brain which have not been previously identified with other AVP ligands. In addition, testicular hormones were found to alter [125I]LinAntag binding in two distinct regions, the posterior lateral preoptic-anterior lateral hypothalamic continuum and the posterior ventrolateral hypothalamic nucleus and adjacent tuberal area.

It has been previously shown that chronic water deprivation or hypertonic saline intake, osmotic stress models with concomitant decrease in food intake, decrease hypothalamic CRH mRNA levels and ACTH responses to acute stimulation. To determine the contribution of food restriction to the effects of osmotic stimulation, the function of the hypothalamic pituitary adrenal axis was analyzed in rats subjected to food deprivation, water deprivation or their combination for 60 h. In all three groups, basal levels of plasma corticosterone were increased, while ACTH and catecholamines were unchanged. Basal plasma vasopressin levels were normal in food deprived rats, but significantly increased in water deprived and simultaneously food and water deprived rats. In contrast to the 25% reduction of plasma ACTH responses to 30 min immobilization by water deprivation, food deprivation had no inhibitory effect and prevented the decreased ACTH responsiveness caused by water deprivation. In control rats, plasma corticosterone levels increased 22.5-fold 30 min after immobilization, and this response was significantly potentiated in the water deprived, food deprived and combined food and water deprived groups. The elevation in plasma catecholamines in response to acute immobilization was also enhanced in both water deprived and food deprived rats. In situ hybridization studies showed a 35% increase in VP mRNA levels in the PVN after water deprivation, whereas food deprivation caused a slight decrease and prevented the stimulatory effect of water deprivation. CRH mRNA in the PVN was reduced by 27% after food deprivation and by 67% after water deprivation, but simultaneous food and water deprivation caused a significantly smaller reduction similar to that in food deprivation alone.(ABSTRACT TRUNCATED AT 250 WORDS)

Two subtypes of arginine vasopressin (AVP) receptors (V1 and V2) have been distinguished. In this study, we examined the characteristics of AVP binding in rat hippocampus and the effects of bilateral adrenalectomy and adrenal steroids on its [ 3 H]AVP binding. [ 3 H]AVP binding to rat liver and the hippocampal membranes was strongly inhibited by the V1 antagonist, OPC-21268. ADX resulted in a significant decrease in theB max of AVP binding in the hippocampus. Chronic treatment with aldosterone and corticosterone restored the ADX-induced reduction, but treatment with dexamethasone did not. These results suggest that the AVP V1 receptor in the hippocampus is regulated by adrenocortical neuroregulatory function.

We investigated the effects of vasopressin-related neuropeptides on the hypoxia/hypoglycemia (ischemia)-induced decrease of the CA1 presynaptic potential elicited by stimulation of Schaffer collaterals in rat hippocampal slices. Treatment with arginine-vasopressin (AVP) potentiated the ischemic decrease of the CA1 presynaptic potential. In contrast, a V1 receptor antagonist produced a dose-dependent neuroprotective effect, whereas a V2 receptor antagonist had no effect. The AVP-induced decrease of the CA1 presynaptic potential was completely blocked by simultaneous application of the V1 receptor antagonist. Because AVP4-9 is regarded as the major proteolytic product of AVP in the rat brain, we examined its effect on the ischemic decrease of the CA1 presynaptic potential. Treatment with AVP4-9 produced a more marked reduction of the potential than treatment with AVP itself. The present study demonstrates that stimulation of the V1 receptor has a detrimental effect on the development of ischemic damage whereas V1 receptor blockade has a neuroprotective effect, suggesting that AVP may potentiate ischemic neuronal deficits via V1 receptor stimulation.

To determine central cardiovascular effects of arginine vasopressin (AVP) in the dog, and the nature of the receptors involved, blood pressure (BP) and heart rate (HR) responses were monitored in 18 conscious dogs subjected to third ventricle (i.c.v.) infusions of either AVP or AVP analogs with agonistic or antagonistic properties towards V1, V2 and putative ‘V3’ receptors. Significant blood pressure (BP) icreases were elicited by i.c.v. infusions of: (i) AVP at a rate of 0.01, 1 and 100 ng/min; (ii) the selective V1 agonist (F180, Ferring) at a rate of 1 and 100 ng/min, and (iii) the selective V1 antagonist, dEt2 Tyr(Me)DAVP at a rate of 100 ng and 400 ng/min. Pretreatmetn with anothe selective V1 antagonist (MeCAAVP) increased baseline BP and prevented AVP induced BP increase. Blood pressure was not altered by i.c.v. infusions of the selective V2 agonist dVDAVP, the selective V2 antagonist d(CH2)5[ d -Ile2,Abu4]AVP, the putative ‘V3’ agonist agonist vasopressin-(4–8) (Akzo Organon), mediating behavioral actions of AVP and by artificial CSF. Heart rate was significantly accelerated by AVP infused at a rate of 100 3g/min. The results reveal high sensitivity of teh conscious dog to centralpresor action of AVP and indicate that this effect is mediated by V1-like receptors. It is suggested that the pressor of the V1 antagonist may result from its partial agonistic properties towards central V1 receptors, however, it can not be excluded that endogenous AVP released under baseline conditions may exert tonic hypotensive effect mediated by a different population of V1 receptors, this effect being abolished by V1 antagonist. The results do not support involvement of V2 or ‘V3’ receptors in central cardiovascular actions of AVP.

The luteinizing hormone surge in the female rat is not only induced by the positive feedback of estradiol, but also by circadian signals originating in the suprachiasmatic nucleus (SCN). In a previous study we showed that administration of vasopressin, an SCN transmitter present in preoptic projections, induced an LH surge in animals bearing complete lesions of the SCN. This strongly suggests vasopressin as a stimulatory circadian signal for the timing of the LH surge. In the present study we investigated during which time window vasopressin may act in the medial preoptic area to stimulate LH secretion in SCN-intact female rats. Vasopressin or a specific V1a receptor antagonist was administered into the MPO by a reverse microdialysis technique during different time windows, and plasma LH concentrations were measured. Vasopressin stimulated the LH surge in 30% of the animals, when administered during the second half of the light period, but during the first half of the light period no effects were observed. Administration of the V1a receptor antagonist, however, did not affect the LH surge. These data confirm our previous results that vasopressin is a stimulatory factor for the LH surge also in SCN-intact animals, and indicate that a certain time window is available for such stimulation. We hypothesize that vasopressin in the SCN-intact animal may act as a circadian signal during a specific time window to induce the LH surge. The time window is the result of other SCN regulatory systems that are involved in the preparation of the LH surge.

Oxytocin facilitates maternal behaviour in sheep. In the present study, we searched for the presence of oxytocin and vasopressin binding sites in the sheep olfactory bulb, a brain area which is thought to be involved in specific bond formation between the ewe and its lamb. Using in vitro autoradiography, we observed binding of tritiated vasopressin to the glomerular layer of the olfactory bulb. Competition studies performed with structural analogues and the use of a 125I-labelled linear vasopressin antagonist suggested that sites which bind vasopressin are V1 type receptors. In contrast, specific binding sites for oxytocin in the olfactory bulb could be detected neither in control females, nor in ovariectomized females treated with estradiol nor in postparturient ewes, although such sites were present in the uterus.

Using autoradiography on film, specific binding sites for arginine-vasopressin (AVP) and for oxytocin (OT) were localized in various areas of the brain of adult male guinea pigs. Vasopressin binding sites were detected with [3H]AVP or with [125I]VPA, a recently synthetized linear vasopressin antagonist radiolabeled with 125I. [125I]VPA and [3H]AVP yielded similar results, thus suggesting that AVP binding sites present in the guinea pig brain are V1 type receptors. Supporting evidence on this was obtained in competing studies using structural analogues allowing to discriminate V1 receptors from V2 and from OT receptors. Oxytocin binding sites were labeled with [3H]OT or with the iodinated OT antagonist [125I]OTA; both ligands yielded similar results. The localization in the guinea pig brain of AVP binding sites differed from that of OT binding sites. AVP binding sites were mainly detected in the olfactory bulb and throughout the cerebral cortex. Oxytocin binding sites were most noticeable in the hypothalamic ventromedial nucleus, in the amygdaloid complex and in restricted areas of the cerebral cortex. A comparison of the present data with those previously described in the rat, the mouse, the human and the hamster brain suggests that similar binding sites are present in these species, but that their anatomical distribution differs markedly. These data are discussed in relation to immunocytochemical and electrophysiological data which suggest that binding sites detected by autoradiography may represent, at least in part, functional neuronal receptors.

The purpose of the present study was to determine the overall cardiovascular and sympathetic nervous system responses to stimulation of neuronal cell bodies in the paraventricular neucleus (PVN) of the hypothalamus. Bilateral microinjections (50 nl) of monosodium glutamate or sodium acetate were made into the PVN of conscious unrestrained rats. Blood pressure, heart rate and plasma concentrations of norepinephrine and epinephrine were measured. The injection of sodium acetate as an osmotic control was without effect on any of the recorded variables. In contrast, the injections of glutamate were associated with a rapid increase in both blood pressure and heart rate. At doses of 15, 25, and 50 nmol blood pressure increased by13 ± 2, 14 ± 3and16 ± 1mmHg while heart rate increased by64 ± 15, 73 ± 8 and50 ± 8 bpm. These responses were associated with increases in plasma norepinephrine concentrations of51 ± 8, 100 ± 16and62 ± 13pg/ml while epinephrine concentrations rose by42 ± 17, 58 ± 18and38 ± 17pg/ml. The responses of glutamate (25 nmol) were not affected by blockade of vascular vasopressin receptors with d(CH 2 ) 5 Tyr(Me)AVP (10 μg/kg) (blood pressure: pre5 ± 3vs post13 ± 3mmHg, heart rate pre77 ± 9bpm vs post91 ± 7bpm, plasma norepinephrine: pre106 ± 22vs post121 ± 28pg/ml and plasma epinephrine: pre61 ± 25vs post34 ± 30pg/ml. These findings indicate that stimulation of neuronal cell bodies in the PVN of conscious rats causes an increase in sympathoadrenal outflow resulting in an increase in blood pressure and heart rate. Vasopressin release into the systemic circulation does not appear to play a major role in the cardiovascular responses associated with PVN stimulation.

The effect of arginine vasopressin (AVP) on its own septal release was evaluated using an in vitro superfusion procedure. As compared to basal release from septal fragments, pulses of synthetic AVP (15 pg/5 min) resulted in a 25-fold augmented release of endogenous AVP, indicating a positive feedback action. Both the basal and stimulated AVP release were significantly increased by 60 mM potassium and markedly reduced by omission of calcium. Preincubation of the septal fragments with the V 2 /V 1 AVP receptor antagonist d(CH 2 ) 5 [ d -Tyr (ET) 2 , Val 4 ]AVP resulted in a dose-dependent inhibition of the positive feedback action of AVP which was nearly completely blocked at doses between 1.25 and 5 ng per 100 μl incubation medium. As compared to this effect, the V 1 antagonist d(CH 2 ) 5 Tyr (Me) 2 AVP as well as oxytocin were significantly less potent. The results suggest that the positive feedback action of AVP on its own release from septal fragments is potassium-stimulated, calcium-dependent and mainly V 2 receptor-mediated. The physiological significance of this phenomenon remains to be shown.

The neuropeptide arginine vasopressin modulates neuroadaptive processes, including memory consolidation and functional tolerance to ethanol, by actions at CNS V1 receptors. Noradrenergic systems play a role in these actions of the peptide. To assess whether vasopressin may act presynaptically on catecholamine neurons, vasopressin receptors were measured by quantitative autoradiography in the lateral septum, an area that is innervated by catecholaminergic neurons and has a high density of V1 receptors, of control and 6-hydroxydopamine-treated mice. Vasopressin receptors were distributed non-uniformly throughout the lateral septum, with greater binding in the more caudal regions. Treatment with 6-hydroxydopamine lowered septal catecholamine levels and vasopressin binding, with a greater effect on binding in the intermediate and caudal portions of the lateral septum. Pretreatment with desmethylimipramine reversed the depletion of norepinephrine, and attenuated the effect of 6-hydroxydopamine on vaspressin binding in the intermediate region, but was less effective in the caudal region of the lateral septum. The results suggest that a portion of septal vasopressin receptors are localized on the terminals of noradrenergic and, possibly, dopaminergic neurons, consistent with the hypothesis that certain neuroadaptive responses to vasopressin could be mediated by modulation of neurotransmitter release. In contrast to the results with 6-hydroxydomapine, treatment of mice with 5,7-dihydroxytryptamine, to destroy serotonergic terminals, did not alter vasopressin binding in the lateral septum.

The intracerebroventricular administration of arginine vasopressin suppressed significantly the fever evoked by interleukin-1. This antipyretic action of arginine vasopressin was blocked completely by the antivasopressor analog d(CH2)5Tyr(Me)arginine vasopressin, an antagonist of the V1 subtype of peripheral vasopressin receptor. However, in contrast to AVP, the V2 receptor agonist, 1-desamino-8-D-arginine vasopressin, did not alter the normal time course or magnitude of interleukin-1 fever. These data suggest that arginine vasopressin induced antipyresis is mediated via central receptors which may resemble the V1 subtype of peripheral vasopressin receptor. The V2 subtype of vasopressin receptor is unlikely to be involved since an agonist of this receptor did not exhibit any antipyretic activity against interleukin-1 fever.

In brain slices of normal Wistar and Long-Evans rats, brief high frequency stimulation of the fimbria fibers induced long-term potentiation (LTP) in excitatory transmission between these fimbria fibers and neurons and teral septum (LS). Slices prepared from diabetes insipidus (DI) Brattleboro rats, that contained no vasopressin (VP), consistently failed to maintain LTP in this excitatory transmission. Exogenous VP, administered to slices from DI Brattleboro rats shortly prior to the experiment or released from a subcutaneous depot in DI Brattleboro rats for several days prior to decapitation, corrected this failure. The maintenance of LTP in the LS in slices from Wistar and Long-Evans rats was prevented by d (CH2)5-Tyr(Me)-arginine VP, an antagonist for the V1 type of VP receptors. These results indicate an important role of VP in the maintenance of LTP in excitatory transmission in the LS. It is conjectured that the effects of VP on LS neurons are related to the role of the peptide in the maintenance of LTP and that these processes play a role in memory formation.

Following two weeks of cerebroventricular administration of arginine-vasopressin (AVP) by Accurel implants, two types of binding sites for this peptide were immunocytochemically visualized in blood vessels in the brain of Brattleboro (di/di) rats: (1) endothelial cells of capillaries were stained with the highest density in hippocampus, striatum, and locus coeruleus (LC), whereas only few such stained cells were present in the septum and cerebral cortex. (2) Bound AVP was also present on endothelial cells and pericyte-like cells in larger blood vessels in striatum and the LC. Both types of vasopressin binding site staining on blood vessels were dose-dependent and could be further enhanced by additional in vitro preincubation of the fixed sections with AVP. This staining was not present in rats implanted with either oxytocin or alpha-melanocyte-stimulating hormone.

We have investigated whether microinjectuions of arginine-vasopressin (AVP) into the nucleus tractus solitarii (NTS) of rats results in the stimulation of inositol phospholipid metabolism, a biochemical event related to the activation of V1-type receptors. We found that AVP, but not the related peptides oxytocin or deamino- d -AVP, increases the incorporation of [3H]myoinositol into lipids and the accumulation of labelled inositol phosphates in the dorsomedial medulla oblongata of lithium-treated rats. These effects were abolished by local pretreatment with d -(CH2)5-Tyr(Me)-AVP, a specific antagonist of V1-receptors. It is concluded that the effects of AVP in the NTS of rats are likely to be due to an action on specific receptors (V1-like) and subsequent activation of polyphosphoinositide turnover.

The paraventricular nucleus of the hypothalamus (PVN) is composed of magnocellular and parvocellular subdivisions. Magnocellular neurosecretory neurons project to the neurohypophysis while parvocellular neurons send monosynaptic axonal projections to autonomic regulatory areas in the brainstem and spinal cord. In the present study, we investigated the hemodynamic effects produced by selective magnocellular or parvocellular stimulation. In urethane anesthetized rats with intact baroreflexes, magnocellular and parvocellular stimulation produced only slight differences in hemodynamic responses, however, following acute sinoaortic denervation a clear difference was observed. Parvocellular stimulation produced an increase in arterial pressure and vasoconstriction in gut, kidney and skeletal muscle. Magnocellular stimulation produced little effect on arterial pressure and marked vasodilation in the hindquarters. Blockade of peripheral vasopressin vascular receptors did not affect the vasoconstrictor response produced by stimulation of PVN. These data are consistent with the hypothesis that the long descending neural projections of the parvocellular PVN subserve a selective vasoconstrictor function.

Vasopressin is one of numerous neuropeptides contained in sympathetic ganglia, but whose function remains unresolved. In this report, we present electrophysiological evidence that arginine-vasopressin (AVP) is a neurotransmitter in guinea pig inferior mesenteric ganglion (IMG). AVP superfused over the IMG, in vitro, produced in a population of neurons a membrane depolarization accompanied by a resistance increase, both of which were blocked by a specific V1 receptor antagonist. Moreover, slow excitatory postsynaptic potentials (EPSPs) elicited by repetitive nerve stimulation were attenuated in 75% of cells tested in the presence of excess AVP, and occasionally in the presence of the antagonist. Thus, AVP joins substance P as a putative transmitter of slow potentials in the guinea pig IMG.