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Blocking effects of OPC-21268 and OPC-31260 (vasopressin V1- and V2-receptor antagonists) on vasopressin-induced constrictions in isolated, perfused dog femoral arteries.

Using a perfusion technique of isolated vessels, constrictor responses to vasopressin (VP) and norepinephrine (NE) were investigated in perfused dog femoral arteries. Both OPC-21268, a selective V1-antagonist, and OPC-31260, a selective V2-antagonist, significantly shifted the VP-induced dose response curves to the right without influencing the NE-induced ones. The blocking effects of OPC-31260 were much greater than those of OPC-21268, suggesting that there may probably be functional V1- and V2-receptors in isolated dog femoral arteries that mediate vasoconstriction.

Angiotensin Receptor Antagonists↗

Adaptation to sustained high plasma vasopressin in water and electrolyte homeostasis in the rat transgenic for the metallothionein-vasopressin fusion gene.

Prolonged exposure of tissues to a receptor agonist often leads to adaptive changes that limit the subsequent responsiveness of the tissue to the same agonist. Recently, we have generated rats transgenic for the metallothionein I-human arginine vasopressin (AVP) fusion gene (Tg), which produced high plasma AVP with relatively preserved renal water excretion, suggesting that there might be adaptive mechanism(s) for maintaining water and electrolyte homeostasis against chronic AVP oversecretion from the earliest stage of life. In this study, to investigate whether down-regulation of AVP V2 receptor (V2R), which could possibly be caused by long-standing high plasma AVP, participates in this adaptive mechanism(s), non-peptidic V2R antagonist OPC31260 was administered to reverse the down-regulation, and water loading was performed after V2R antagonist treatment had been withdrawn. Additionally, to confirm the down-regulation, Northern blotting analysis for V2R mRNA was carried out. Tg rats showed slightly decreased urine volume and water intake with an equivalent plasma [Na(+)] level (Tg 140.4 +/- 0.6 mEq/l; control 139.3 +/- 0.6 mEq/l) under basal conditions. After water loading using a liquid diet containing zinc, which stimulates the promoter region in the transgene, the urine increase showed only limited suppression with a dramatically increased plasma AVP level and mild hyponatremia (135.8 +/- 1.8 mEq/l) in Tg rats. When diet containing OPC31260 had been provided for 4 days until the day before the start of water loading, antidiuresis and hyponatremia (125.4 +/- 1.mEq/l) were significantly potentiated. V2R mRNA expression in kidney was significantly less in Tg rats than in control rats under basal conditions, and this suppression was restored by OPC31260 treatment to levels comparable with those of control rats. These results suggest that long-standing high plasma AVP causes V2R down-regulation, and it may play an important role in the adaptive mechanism(s) for maintaining water and electrolyte homeostasis in chronically AVP-overexpressing rats.

Adaptation, Physiological↗

A novel arginine vasopressin-binding peptide that blocks arginine vasopressin modulation of immune function.

We report on a novel peptide that blocks the neuroendocrine hormone arginine vasopressin (AVP) helper signal for IFN-gamma production by direct interaction with the hormone. The AVP-binding nonapeptide has the sequence Thr-Met-Lys-Val-Leu-Thr-Gly-Ser-Pro (binding peptide). AVP and its 6-amino acid N-terminus cyclic ring pressinoic acid (PA) are both capable of replacing the IL-2 requirement for IFN-gamma production by mouse splenic lymphocytes. We show that the AVP-binding peptide specifically and reversibly blocks AVP help in IFN-gamma production, but fails to block the helper signal of PA. Thus the intact AVP molecule and not just the N-terminal cyclic ring is important for interaction with the binding peptide. AVP interacts with the binding peptide with an apparent KD of approximately 50 nM. The AVP-binding peptide does not inhibit AVP interaction with its receptor on lymphocytes. Interestingly, whereas the AVP-binding peptide does not block the PA helper signal for IFN-gamma induction, the complex of AVP and binding peptide does reversibly block the PA signal. The AVP family of hormones requires conformational flexibility for signal transduction. Thus, we hypothesize that the AVP-binding peptide restricts this flexibility and converts AVP into an antagonist of its own action.

Adjuvants, Immunologic↗

1-deamino-/4-valine-8-D-arginine/-vasopressin (dVDAVP), a new synthetic vasopressin analog for treating diabetes insipidus.

The effects of LVP, dDAVP, and dVDAVP on the water excretion of 14 vasopressin-sensitive and 2 ADH-resistant patients with diabetes insipidus were studied and compared. Findings showed that dDAVP and dVDAVP decreased the diuresis of all ADH-sensitive patients, and in those with diabetes insipidus, more markedly (7.6-fold) and for a longer duration (3.3-fold) than did LVP. From a comparison of the results, dVDAVP is at present one of the most effective preparations for reduction of polyuria in patients with ADH-sensitive diabetes insipidus.

Adult↗

V1 vasopressin receptor antisense oligodeoxynucleotide into septum reduces vasopressin binding, social discrimination abilities, and anxiety-related behavior in rats.

To develop and validate a vasopressin (AVP) receptor knockdown strategy, we infused an antisense oligodeoxynucleotide to the V1 subtype mRNA into the septum of male rats with osmotic minipumps and measured behavioral, cellular and molecular parameters. Compared to vehicle and scrambled-sequence oligo controls, chronic antisense administration for up to 4 d diminished the ability of the animals to distinguish a previously exposed juvenile from a novel one and to respond to exogenous AVP (1 ng/5 microliters, intracerebroventricular) with an improved social memory. Furthermore, anxiety-related behavior was reduced. As measured in the behaviorally tested rats, antisense treatment resulted in a reduced binding of radiolabeled AVP in the septum, but not in other limbic brain areas (receptor autoradiography), and an increased amount of V1 receptor mRNA (reverse transcriptase PCR), indicating translational arrest and ongoing transcriptional activity. In sense oligo-treated rats, on the other hand, both the social and the anxiety-related behavior scores lay between levels obtained in control and antisense-treated animals. These sense-treated rats showed a slightly reduced V1 receptor density in the septum and reduced receptor mRNA levels, indicating hybridization of the sense oligo to the DNA. The data show the potential of antisense targeting to further reveal relationships between local gene expression, neuropeptide-receptor interactions in distinct brain areas, and behavioral performance.

Analysis of Variance↗

Vasopressin and galanin mRNAs coexist in the nucleus of the horizontal diagonal band: a novel site of vasopressin gene expression.

Vasopressin (VP) neurons have been identified in several brain regions where VP has been hypothesized to act as a neurotransmitter or neuromodulator. In many sites, VP is colocalized with the neuropeptide galanin (GAL). Here, using single in situ hybridization histochemistry, we have identified a novel group of neurons within the nucleus of the horizontal diagonal band of Broca (HDB) that express the VP gene and have assessed the distribution of these cells in adult male and female rats (90 days old, n = 7/group). VP mRNA-expressing neurons were scattered throughout the rostrocaudal extent of the HDB, and the number of VP neurons detected unilaterally ranged from 1 to 17 cells per 20 microns section. Using double in situ hybridization histochemistry on alternate sections, we have assessed the number of cells expressing VP and/or GAL mRNA in the diagonal band and have determined the extent of their colocalization. Approximately 50% of all VP-expressing neurons in the HDB coexpressed GAL mRNA, and 33% of GAL-expressing neurons in this region coexpressed VP mRNA. No sex differences were detected in the number of neurons expressing either VP or GAL mRNA or in the incidence of coexpression of VP and GAL mRNAs in this region. VP neurons in the HDB exhibited a low level of expression, and cellular VP mRNA content did not differ between male and female rats. However, sex differences were present in the bed nucleus of the stria terminalis (BNST) of these same rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Distinct inhibition by non-peptide and peptide arginine vasopressin antagonists of vasopressin-induced activation of mitogen-activated protein kinase in cultured rat vascular smooth muscle cells.

The preincubation of vascular smooth muscle cells (VSMC) with a non-peptide arginine vasopressin (AVP) antagonist (OPC-21268) (0.5 mM) for 10 min completely inhibited the 1 microM AVP-induced activation of mitogen-activated protein (MAP) kinase but did not affect the angiotensin II-, protein kinase C activator- or ionomycin-stimulated MAP kinase. Similar results were obtained with the simultaneous administration of AVP and OPC-21268. This inhibitory effect of OPC-21268 completely disappeared after washing the cells pretreated with OPC-21268. In contrast, the preincubation of VSMC with a peptide AVP antagonist, d(CH2)5Tyr(Me)AVP (1 microM), for 10 min completely blocked the 1 microM AVP-activated MAP kinase but the simultaneous administration of d(CH2)5Tyr(Me)AVP with AVP could inhibit that only by 40%. However, the washing procedure did not affect the inhibitory effect of d(CH2)5Tyr(Me)AVP. These results indicate that OPC-21268 is a specific AVP antagonist to inhibit the AVP-induced activation of MAP kinase. The antagonistic effect of OPC-21268 is also suggested to be relatively weak but prompt as compared to that of a peptide AVP antagonist d(CH2)5Tyr(Me)AVP.

Angiotensin II↗

Increased vasopressin secretion from hypothalamic cultures following administration of exogenous vasopressin mRNA.

In Brattleboro rats, exogenous vasopressin (VP) mRNA can be accumulated, transported, and translated by magnocellular neurons. To determine whether this phenomenon may also occur in magnocellular neurons of normal rats, dispersed hypothalamic neuronal cultures of fetal Sprague-Dawley rats were exposed to VP mRNA. The cultures were maintained in either control medium or medium containing the cAMP elevating drugs, IBMX (3-isobutyl-1-methylxanthine), and forskolin for 23 or 30 days of culture to induce VP synthesis and secretion. Following removal of the IBMX and forskolin at Day 23, VP secretion into the medium declined to baseline by 30 days in vitro, but administration of VP mRNA to these cultures on Day 30 resulted in a 5-fold increase in VP content of the medium (P = 0.005) after 6 h and a 2.5-fold increase after 24 h (P = 0.002). Administration of VP mRNA to the cultures treated continuously with IBMX and forskolin also resulted in a small increase in VP secretion which did not reach significance after either 6 or 24 h. When cultures prepared with continuous I/F were exposed to antisense VP mRNA, VP secretion into the media was decreased by 58%, and VP immunoreactive perikayra were difficult to observe. This demonstrates that the increase in VP release observed after the addition of sense VPmRNA did not reflect a nonspecific effect of the addition of mRNA to the culture medium. Autoradiography of cultures administered 3H- or 32P-VP mRNA for 24 h revealed silver grains associated with varicosities, perikarya, and neuritic processes of neurophysin (NP)-positive, but not NP-negative neurons. These results suggest that exogenously administered mRNA has access to cell translation systems in cultured hypothalamic neurons as well as magnocellular neurons of Brattleboro rats.

Animals↗

Effects of vasopressin, des-glycinamide vasopressin and amphetamine on a combined passive and active avoidance task.

The effects of peripherally injected arginine vasopressin (AVP), its des-glycinamide derivative (DGAVP), which has limited pressor activity, and d-amphetamine (AMP) were studied on a combined passive and active avoidance task (Carew 1970). In this procedure, the rat experiences mild foot-shock in one of two distinctively coloured compartments, followed by drug treatment. On the next day, re-entry latency into either of the two compartments is measured (latency index), and the actual compartment chosen is noted (choice index). Both measures were affected by the intensity of the shock. A moderate dose of AVP (25 micrograms/kg) increased latencies, but only when a relatively low shock intensity was used; choice was not markedly affected. DGAVP did not significantly affect performance under any condition. AMP affected latency scores as well as choice behaviour at the lower shock level but, presumably due to "ceiling" effects, did not exert an effect in the high shock condition. The negative DGAVP findings suggest that the behavioural potency of peripherally injected AVP was due to pressor activity, rather than any direct action on cognitive mechanisms. Moreover, this indirect AVP effect on mnemonic performance was relatively weak, since only the latency, but not choice, measure was significantly affected. It is concluded that the Carew paradigm provides a more comprehensive assessment of behaviour than traditional passive or active avoidance procedures.

Animals↗

Contrasting effects of vasopressin, desglycinamide-vasopressin and amphetamine on a delayed matching to position task in rats.

The effects of peripherally injected arginine vasopressin (AVP: 0-25 micrograms/kg), its desglycinamide analogue (DGAVP: 0-25 micrograms/kg), which is practically devoid of pressor activity, and d-amphetamine (AMP: 0-1.25 mg/kg) were studied using a delayed (0-32 s) matching to position task (Dunnett 1985). A limited hold for responding (20 s) was in operation. This task enables an accurate assessment of forgetting in rats. AVP reliably improved per cent correct performance, and this effect was substantiated by accuracy indices derived from signal detection theory (TSD). DGAVP, however, was inactive, suggesting that the parent peptide's pressor properties were responsible for its beneficial effects. AMP disrupted performance in a dose-related manner, and was the only substance to alter a TSD bias index (responsivity index, RI), indicating a degree of response repetition at the highest dose. These results are consistent with some earlier reports, and suggest that AVP may enhance memory by peripheral action, while AMP disrupts performance. Closer inspection of the data, however, suggested that the peptide reduced general responsiveness. A new index to measure bias (Sahgal 1987) suggested that AVP-treated subjects restricted their sample, and choice responses to one side of the operant chamber, thereby achieving a spuriously high detection rate with few errors of commission (incorrect responses). It is concluded that AVP does not, after all, improve performance: on the contrary it has detrimental effects, and produce errors of omission (failure to respond).

Animals↗

Regulation of vasopressin gene expression: changes in the level, but not the size, of vasopressin mRNA following endocrine manipulations.

1. Regulatory interactions between the hypothalamoneurohypophyseal vasopressin (VP) axis and the endocrine systems of the anterior pituitary have been investigated in the rat by observing changes in VP mRNA expression following endocrine manipulations. 2. An increase in the level, but not size, of VP mRNA was found in the supraoptic (SON) and paraventricular nuclei (PVN) of the hypothalamus and in the neurointermediate lobe (NIL) of the pituitary following hypothyroidism (induced by drinking 6-n-propyl-2-thiouracil; PTU) and adrenalectomy. Hypothyroidism induced by alternative procedures (surgical thyroidectomy or PTU injections) did not exert similar effects. 3. Treatment with the dopamine agonist bromocriptine to reduce prolactin secretion raised levels of VP mRNA in the NIL only. Castration did not up-regulate VP mRNA levels. 4. Since the observed effects on VP mRNA levels occur in the absence of changes in plasma osmolality, these results provide evidence of nonosmotic regulation of VP gene expression, an effect which is observed most clearly in the NIL pool of VP mRNA. Furthermore, the effects are distinct from changes in VP mRNA levels associated with raised plasma osmolality since the VP mRNA size was not increased.

Adrenalectomy↗

Role of blood osmolality in the regulation of vasopressin secretion in man: application of a new radioimmunoassay method for vasopressin.

A radioimmunoassay for plasma arginine vasopressin (AVP) has been developed based on R2 antibody of Thomas and Lee, synthetic standard (Ferring) and extraction on Sep-Pak column. High recovery of AVP (approximately 79%) was achieved with a detection limit of 0.25 pg/ml. By improving the technique of measurement of plasma osmolality an intraassay coefficient of variation less than 1% was obtained. Physiological studies performed with this method demonstrated that AVP becomes undetectable after water loading 20 ml per Kg of water po; (N = 6) and increases in response to hypertonic saline infusion (0.05 ml/kg/min; N = 15) with a linear relationship between plasma osmolality and AVP in individual subjects; this relationship is maintained when the test is repeated in the same subjects. However when pooling all data together, the relationship between plasma osmolality and AVP is best expressed by an exponential relationship. This implies that after AVP release is initiated, the concentration of the hormone increases more rapidly than plasma osmolality and the release is continuous possibly due to recruitment of increasing number of neuronal units whose osmotic threshold varies from individual to individual.

Administration, Oral↗

Extrahypothalamic vasopressin and oxytocin in the human brain; presence of vasopressin cells in the bed nucleus of the stria terminalis.

In the present study, the distribution of extrahypothalamic vasopressin (VP) and oxytocin (OXT) in the human brain was investigated by means of immunocytochemistry. In the septum verum, few VP fibers were found in the nucleus septalis lateralis and medialis (NSL and NSM), and in the bed nucleus of the anterior commissure. Very few VP and OXT fibers were present in the amygdala and in the hippocampus, mainly around the rostral tip of the lateral ventricle on the level of the pes hippocampi. The locus coeruleus (LC) contained dense networks of VP fibers and, although to a lesser extent, OXT fibers over its entire rostrocaudal extension. VP-immunoreactive neurons were present in the bed nucleus of the stria terminalis in a number of subjects, while no OXT cells were found in this structure. Thus, the VP innervation of limbic structures in the human brain, in particular of the NSL, was found to be clearly less pronounced than in the rat brain. The VP innervation of the LC, by contrast, was denser in the human brain than in the rat brain. No sex differences were found in the VP innervation of the human brain. These findings stress the need for caution in extrapolation of data concerning peptidergic innervation of the rat brain towards the human brain.

Adolescent↗

Microinjection of anti-vasopressin serum into hippocampus in mice: effects on appetitively reinforced task after intraventricular administration of Arg-vasopressin.

Antiserum to [Arg8]vasopressin (anti-AVP) was bilaterally administered into dorsal hippocampus at 1:50 or 1:10 dilution 20 min before the 24-day retention session of a visual discrimination task. This treatment by itself did not affect the retention performance by comparison with the respective control group, whatever the dilution of anti-AVP, suggesting that hippocampal endogenous AVP is not involved in our behavioral paradigm. On the other hand, intracerebroventricular (i.c.v.) administration of AVP 10 min before the retention session improved retention performance of the visual discrimination task. When anti-AVP was injected at the 1:10 dilution into the dorsal hippocampus 10 min before the i.c.v. administration of AVP, the retention performance was not improved. These data suggest the involvement of the hippocampus in the behavioral expression of AVP following an i.c.v. treatment.

Animals↗

Arginine vasopressin-binding peptides derived from the bovine and rat genomes differ in their abilities to block arginine vasopressin modulation of murine immune function.

Previously we reported on a nonapeptide (binding peptide) derived by reading the complementary DNA strand of the bovine arginine vasopressin (AVP) gene in the 3'5' direction that specifically blocks the AVP helper signal for gamma-interferon (IFN gamma) production by helper cell-depleted mouse spleen cultures. Bovine 5'3' AVP-binding peptide, however, did not block AVP activity. We report here on the relative abilities of 5'3' and 3'5' AVP-binding peptides derived from the bovine and rat AVP genes to block the AVP helper signal for IFN gamma production. The sequences of the bovine and rat 5'3' AVP-binding peptides differ by two amino acids, whereas the 3'5' AVP-binding peptides derived from both genes are the same. In contrast to the lack of blocking activity of the bovine 5'3' AVP-binding peptide, the rat 5'3' AVP-binding peptide was almost as effective as the 3'5' AVP-binding peptide in blocking AVP function. No effect was seen with a 9-amino acid control peptide consisting of a 'scrambled' 3'5' AVP-binding peptide sequence. Further, polyclonal anti-rat 5'3' AVP-binding peptide antibodies blocked AVP activity, whereas polyclonal anti-bovine 5'3' AVP-binding peptide antibodies had no significant effect. Polyclonal antibodies generated against the 3'5' AVP-binding peptide also blocked AVP activity. These antibodies possibly blocked AVP function by interaction with the lymphocyte AVP receptor, since AVP specifically displaced binding of the antibodies.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

Intracerebroventricular application of a vasopressin antagonist peptide prevents the behavioral actions of vasopressin.

Three experiments were conducted to test the hypothesis that the memory-improving properties of peripherally-applied vasopressin (AVP) were related to its aversive (i.e. arousing) actions. The memory effects of AVP were observed in a one-trial food-finding task where non-deprived rats were briefly exposed to a large open field that contained an alcove in which a high-incentive familiar food reward (sweetened milk) was freely available. AVP injections immediately upon removal from the open-field produced faster latencies to refind the alcove (compared to vehicle controls) when tested 48 h later. The aversive actions of AVP were demonstrated in two behavioral assays: (1) a conditioned taste aversion test in which rats learned to avoid a preferred saccharin solution after it had been paired with injections of AVP; and (2) a conditioned place test in which rats learned to avoid a distinctive environment associated with AVP administration. Both the memory and aversive responses to AVP were prevented, in a dose-dependent manner, by immediate pretreatment with intracerebroventricular infusions of the pressor antagonist analog 1-deaminopenicillamine-2-(O-methyl)-tyrosine AVP. The large antagonist doses required to block AVP's behavioral effects suggest that the critical site of action may be far removed from the lateral ventricles. The possibility that AVP-induced improvements in memory result from peripheral arousing actions is discussed.

Animals↗

Changes in vasopressin-converting aminopeptidase activity in the rat pineal gland during summer: relationship to vasopressin contents.

Vasopressin (VP)-converting aminopeptidase (VP-AP) activity and VP contents were measured in single rat pineal glands during the summer of two successive years. The peptidase activity decreased significantly in August. The lowest activity (+/- SEM) of 0.18 +/- 0.02 pmol.hour-1 was recorded on August 14, compared to the basal activity of 0.25 +/- 0.01 pmol.hour-1 in July and September of 1986. The change with similar percentage occurred in the same period of 1987. The specific activity of the enzyme in the crude homogenate, 15,000 g pellet and supernatant fraction of rat pineal glands, exhibited the same pattern of variations. The decrease in peptidase activity coincided with the previously reported dramatic rise in pineal VP content in early August which was confirmed in this series of experiments. Another peptidase, the so-called gamma-endorphin generating endopeptidase (gamma-EGE) activity, and beta-endorphin-related peptides in the pineal gland did not change in this period. The results show that the variations of pineal VP contents and VP-AP activity during summer are not general for other peptides and peptidases. The coincidence of opposite changes in VP content and VP-AP activity of the pineal gland may indicate a role of the peptidase activity to regulate the VP content.

Acclimatization↗

A daily rhythm in behavioral vasopressin sensitivity and brain vasopressin concentrations.

The concentration of Arg-vasopressin (AVP) in the suprachiasmatic nuclei of the hypothalamus (SCN), the neural generator of circadian rhythms, showed a daily rhythm, which was inversely related to the rhythm in lordosis, an aspect of sexual behavior shown by ovariectomized estradiol-17 beta-treated female rats. A threshold dose of an AVP antagonist facilitated sexual behavior most effectively if injected intracerebroventricularly when the endogenous levels of AVP in the SCN were maximal and a threshold dose of AVP inhibited the behavior most effectively if injected when these levels were minimal. The results support the suggestion that AVP may be the neuropeptide whereby the SCN generate some behavioral rhythms.

Animals↗