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Synthesis and biological activities of a photoaffinity probe for vasotocin receptors.

This study reports the synthesis and biological activities of 1-desamino, 7-lysine-(4-azidobenzoyl), 8-arginine vasotocin (d7-N3-AVT). This compound was found to be biologically active in the rat antidiuretic assay (20 U/mg), to behave as an antagonist of vasopressin in the rat pressor assay (pA2 = 6.6), and to yield a half-maximal hydroosmotic response in the isolated toad urinary bladder at a bath concentration of 2.4 X 10(-8) M. When toad bladders were exposed to d7-N3-AVT in the presence of long wavelength UV light, the hydroosmotic response persisted in spite of prolonged and repeated periods of washout. By contrast, the hydroosmotic response in control bladders after stimulation with d7-N3-AVT in the absence of UV irradiation was fully reversed within 15 min of washout. A membrane preparation derived from bladders that had been photolabeled with d7-N3-AVT and washed for 1 h specifically bound 325 fmol [3H]vasopressin/mg protein. Matched bladders exposed to the analog in the absence of UV irradiation and washed for 1 h specifically bound 591 fmol [3H]vasopressin per mg of protein. These studies indicate that d7-N3-AVT binds covalently to hydroosmotic receptors of toad urinary bladder and forms a complex that is functional in triggering an increase in the permeability to water of the epithelium. This analog may prove useful in the isolation and purification of vasotocin receptors in lower vertebrates.

Affinity Labels↗

Arginine vasotocin in the rabbit subcommissural organ.

Subcommissural organs of young and mature rabbits were analyzed for their content of arginine vasotocin by radioimmunoassay. Younger animals had significantly greater quantities of this peptide. There was no detectable arginine vasopressin or oxytocin in subcommissural organ extracts. It is concluded that the subcommissural organ represents, in addition to the pineal and the fetal neurohypophysis, another significant source of arginine vasotocin in the mammalian central nervous system.

Age Factors↗

Expression of hypothalamic arginine vasotocin gene in response to water deprivation and sex steroid administration in female Japanese quail.

Arginine vasotocin (AVT) is a neurohypophyseal hormone involved in reproductive function and control of osmoregulation in birds. In view of the dual function of AVT, the present experiment was designed to observe the effect of water deprivation (WD) and sex steroid [estradiol benzoate (EB) and testosterone propionate (TP)] treatment independently, as well as simultaneously, on the profile/activity of the hypothalamic AVT system. WD resulted in a significant increase in plasma osmolality, sodium ion concentration and AVT concentration, but administration of sex steroids had no significant influence on these parameters. By contrast, the amount of hypothalamic AVT transcript (northern analysis) and the size of immunoreactive vasotocin (ir-AVT) neurons and hybridization signals (in the form of silver grains), representing AVT mRNA in corresponding neurons of paraventricular nuclei (PVN), increased significantly in all the treated groups compared with controls. Our findings indicate that although sex steroid administration has no effect on plasma osmolality and AVT concentration, unlike water deprivation, it may stimulate the profile/activity of AVT neurons of PVN, supporting the possibility of sex steroid receptors on these neurons. It is concluded that in quail, osmotic stress not only upregulates the expression of the AVT gene in existing neurons but also recruits many more neurons to increase the rate of AVT synthesis and secretion, while sex steroids appear to have a stimulatory effect only on the existing number of neurons and only at the level of transcription/translation and hence may influence/modulate hypothalamic AVT gene expression in response to osmotic stress. This study also suggests an interrelationship between reproduction and AVT system/function in birds.

Analysis of Variance↗

The effects of arginine vasotocin on pregnant Mare's serum-induced ovulation in the immature female rat.

Arginine vasotocin (AVT) treatment (1 microgram/injection) every two hours beginning at 06.00 h on the morning preceding expected ovulation significantly inhibited ovulation in 29-day-old immature female rats treated 2 days earlier with 30 IU of PMS. Pre-ovulatory plasma surges and pituitary decreases in LH, FSH and prolactin as measured by radioimmunoassay were not eliminated by AVT treatment. However, there appeared to be an early rise in plasma FSH in these rats and the maximum plasma LH value attained during the surge period in PMS-treated rats was elevated by AVT treatment. Arginine vasotocin significantly inhibited the increase in ovarian weight which occurs in PMS-treated rats on the day preceding ovulation and uterine weights were consistently depressed in AVT-treated rats. The results of this study indicate that AVT likely inhibits PMS-induced ovulation at the level of the ovary, although the possibility cannot be ruled out that inhibition resulted from the advancement of the plasma FSH rise. These data and other studies investigating the effects of AVT on reproduction indicate that AVT is possibly capable of acting at more than one level of the hypothalamo-hypophyseal-gonadal axis.

Animals↗

Guanylylimidodiphosphate enhances the vasopressin- and vasotocin-induced osmotic water flow with no effect on the sodium transport of the frog bladder.

The effects of guanylylimidodiphosphate [Gpp (NH) p] on the vasopressin- and vasotocin-induced osmotic water flow and sodium transport across the bladder of the frog, Rana catesbiana, were examined. Gpp (NH) p enhanced both the vasopressin- and vasotocin-induced water flow, while it did not affect that of the sodium transport. These results suggest that the neurohypophysial hormone-induced water flow is regulated by the guanine nucleotide regulatory site of the adenylate cyclase, while the sodium transport is not regulated at the regulatory site.

Animals↗

Calcium-independent stimulation of glycogenolysis by arginine vasotocin and catecholamines in liver of the axolotl (Ambystoma mexicanum) in vitro.

Arginine vasotocin (AVT) caused a concentration-dependent increase of glycogen phosphorylase alpha activity, breakdown of glycogen and release of glucose, when added to pieces of axolotl liver in organ culture. The concentration causing half-maximal response (EC50) was about 1 nmol/l. These actions of AVT were unaffected by the adrenergic antagonists propranolol, yohimbine and prazosin, but were blocked by equimolar amounts of d(CH2)5Tyr(Me)AVT, a synthetic antagonist of vasopressin. Arginine vasotocin similarly caused glycogenolysis in isolated perfused axolotl liver where the EC50 was about 0.1 nmol/l. The glycogenolytic action of AVT (10 nmol/l) was sustained for at least 3 h in Ca2+-free perfusion and longer in organ culture. No increase in Ca2+ concentration in the effluent perfusion medium was apparent during AVT-induced glucose release. Omission of Ca2+ from the medium, together with addition of EGTA (2.5 mmol/l) to the organ culture, had only a slight inhibitory effect upon the rate of glycogenolysis brought about by AVT and did not inhibit the glycogenolytic action of catecholamines. Addition of the calcium ionophore A23187 (5 mumol/l) neither caused glucose release nor abolished the glycogenolytic action of AVT added subsequently. Nevertheless, A23187 caused increased loss of 45Ca from Ca2+-loaded liver pieces whereas AVT was without effect. There was a slight accumulation of cyclic AMP (cAMP), but not cGMP, in axolotl liver pieces cultured in the presence of 0.1 mumol AVT/l and this was accentuated in the presence of phosphodiesterase inhibitors. We conclude that, in contrast to the position in mammals, Ca2+ is not involved in the glycogenolytic actions of AVT or catecholamines in axolotl liver. Preliminary experiments suggest that the same is true in the carp and we suggest that the involvement of Ca2+ in regulation of hepatic glucose release may not have evolved until after the amphibians separated from the ancestors of the mammals.

Ambystoma↗

Atrial natriuretic factor inhibits vasotocin-induced water reabsorption in the toad urinary bladder.

Peptides recently isolated from atrial extracts possess potent natriuretic and diuretic activities, which are thought to be due to hemodynamic actions, such as increased glomerular filtration or altered medullary blood flow. A direct tubular site of action cannot be ruled out; therefore we have examined the effect of one of these peptides, atriopeptin III on vasotocin-induced water absorption in the toad urinary bladder. Our results indicate that equimolar doses (10(-12) to 10(-11) M) of atriopeptin III can significantly inhibit vasotocin-induced water reabsorption in vitro and suggest a physiologic role for the cardiac peptides to alter water reabsorption directly at the level of the tubules or collecting ducts, independent of any hemodynamic effects they might also exert in vivo.

Absorption↗

Influence of arginine vasotocin on a genetically determined excessive appetite for water in chickens.

Females from two inbred strains of chickens, one normal and one having hereditary diabetes insipidus, were treated with five levels (2.5, 1.0, 0.5, 0.25, and 0.1 units) of Schwartz-Mann (Grade A, 100 units/mg.) arginine vasotocin via ic. injections at 2-hr. intervals. Paired sibs, one normal and one having diabetes insipidus (di), from an F2 population were used in one experiment to compare responses at the level of 2.5 units. Only di females were used for the remaining four levels of treatment. Water intake was determined every two hours and calculated as a percentage of water (ml.)/body wt. Results show that hereditary diabetes insipidus in chickens is arginine vasotocin-sensitive at the 0.5 unit and higher levels. The genetic defect appears to affect the quantity of antidiuretic hormone produced.

Animals↗

[Effect of vasotocin and prostaglandin E2 on absorption of water and ions in the large intestine of a grass fog].

In experiments on the frog Rana temporaria L. isolated colon, it has been shown that the rate of fluid absorption from the lumen is the highest when the lumen contains a hypotonic solution (22.5 mOsm/kg H2O), whereas the rate is lower in case of mucosal Ringer solution (225 mOsm/kg H2O). 10 nmole arginine-vasotocin rises the hypotonic fluid absorption, while 0.1 mumole prostaglandin E2 reduces the fluid absorption. Water content in the colon wall is the highest at mucosal hypotonic Ringer solution. 0.1 mumole prostaglandin E2 decreases hydration of the colon tissue. Secretion of prostaglandins E1 and E2 into the extracellular fluid of the colon has been established. The data obtained indicate participation of vasotocin and prostaglandin E2 in regulation of fluid absorption in the frog colon.

Animals↗

[The effect of injections of arginine vasotocin and mesotocin on the function of the interrenal gland in the frog Rana temporaria 2 months after the removal of the anterior lobe of the hypophysis].

Morphometric and biochemical studies have been made on the activity of interrenal gland in the frog Rana temporaria after removal of the anterior lobe of pituitary and injections of arginine vasotocin and mesotocin. It was demonstrated that the activity of this gland after the operation decreases increasing after arginine vasotocin injections.

Animals↗

The effect of arginine-vasotocin on the production of steroid hormones by mouse, cow, and chicken ovarian tissues in vitro.

The effect of arginine-vasotocin (50 ng ml-1) on the in vitro production of progesterone, testosterone, and oestradiol by isolated cow and chicken ovarian follicles and mouse ovaries was studied. This peptide (1) enhanced the release of progesterone into the medium by the ovarian tissues of all three species of animal; (2) inhibited the production of oestradiol by mouse and cow ovaries; (3) had no significant effect on testosterone secretion in any of the species studied. The results obtained indicate that arginine-vasotocin directly influences steroid production in mammalian and avian gonads.

Animals↗

Synthesis and characterization of a long-acting fluorescent analog of vasotocin.

This study reports the synthesis of l-desamino-7-lysine- (fluorescein)-8-arginine-vasotocin (7-lys(flu) dAVT), and describes its biological activity in the isolated urinary bladder of the toad Bufo marinus. 7-lys(flu)dAVT was fully active in increasing bladder permeability to water. A half-maximal hydroosmotic response was obtained at a concentration of 3 x 10(-8) M. A unique feature of this analog was that its response was not readily reversed after removal of the analog from the serosal bathing solution. The residual response to 7-lys(flu) dAVT was abolished (reversibly) by reducing serosal bath pH from 7.4 to 6.0, suggesting that acidification inhibits the response to analog at a step after the interaction of the ligand with its receptor. Although 8-arginine-vasopressin (AVP) was about 20 times more potent than 7-lys(flu)dAVT in increasing membrane permeability to water, the response to AVP was readily reversed. Preincubation of bladders with 7-lys(flu)dAVT in the presence of AVP blocked the residual response to 7-lys(flu) dAVT. These studies suggest that 7-lys(flu)dAVT forms a stable and physiologically active complex with hydrosmotic toad bladder receptors, and it may, therefore, serve as a useful fluorescent marker for receptors in tissues from this and other species that use vasotocin as an antidiuretic/pressor principle.

Amino Acid Sequence↗

[Morphofunctional research on the interrenal gland of the frog Rana temporaria following arginine vasotocin administration].

Studies have been made of the effect of injections of hypothalamic nonapeptide neurohormone, arginine vasotocin, on functional condition of the interrenal gland in mature frogs. In unoperated, sham-operated and in animals 10 days after hypophysectomy, single and especially three subsequent injections of arginine vasotocin (5 x 10(-9) M per 1 kg of the body weight) result in evident activation of glandular cells of the interrenal gland which is manifested in the increase of the volume of their nuclei and cytoplasmic area, as well as in the dilatation of the blood vessels. Activation of the interrenal gland in hypophysectomized frogs, which lack endogenous ACTH, indicate the direct para-adenohypophyseal influences of nonapeptide hypothalamic hormones on the activity of glandular cells in the peripheral endocrine glands, in particular, the interrenal gland of the grass frog.

Animals↗

[Radio-immunologic determination of hypophyseal vasotocin by Phoxinus laevis during experimental modification of osmotic equilibrium].

Hypophysial vasotocin of Phoxinus laevis is determined by radio-immunoassay. With different sea water dilutions it is shown that hypophysial vasotocin concentration varies with ambient salinity. This fact agrees with histological pictures. Moreover, in comparison with the results of other authors, it is established that the hypophysial morphological modifications appeared only after the hormonal changes.

Animals↗

The effect of hemorrhage on blood arginine vasotocin level in the domestic fowl.

The effect of hemorrhage on blood arginine vasotocin level in the domestic fowl. Acta Physiol. Pol., 1978, 29 (2): 107--110. The blood arginine vasotocin (AVT) level of domestic fowl after hemorrage was determined on the isolated bladder of the frog. It was found that hemorrhage equivalent to 10--20% of the blood volume caused a 2--3 times greater increase in blood AVT level compared to that found before stimulation.

Animals↗

Central cardiovascular effects of vasotocin, oxytocin and vasopressin in conscious rats.

Arg8-vasotocin (AVT), oxytocin and Arg8-vasopressin (AVP) were injected into the i.c.v. space and/or the hypothalamus of awake, freely moving rats in order to investigate the potential role of these neuropeptides in central cardiovascular control. AVT (0.015-10 nmol) injected i.c.v. caused dose-dependent pressor responses; low doses also elicited tachycardia, whereas the higher doses (1 and 10 nmol) produced bradycardia. In contrast, oxytocin injections into the lateral ventricle (0.015-100 nmol) had no significant cardiovascular effects. Plasma vasopressin was not affected by i.c.v. administration of oxytocin or AVT except at the highest dose of AVT. AVT (0.1 or 1 nmol) and AVP (0.015 or 0.1 nmol) injected into the hypothalamic nucleus preopticus medialis (POM) increased blood pressure and heart rate; these changes were much greater than with comparable doses of AVT injected i.c.v. Plasma levels of norepinephrine and epinephrine, but not vasopressin, were significantly elevated during the pressor period induced by 1 nmol of AVT injected into the POM. The pressor and cardiac accelerating effects of AVT (1 nmol) injected into the POM were significantly diminished in bilaterally adrenal demedullated, bretylium-treated rats. These data suggest a role for AVT, but not oxytocin, in central cardiovascular control through activation of the sympathoadrenomedullary axis. The anteroventral hypothalamus might play a central role in these autonomic effects of vasotocin. It is also suggested that AVP and AVT share some central autonomic effects due to similarities in the side-chain of their molecules.

Animals↗

[The effect of vasotocin and mesotocin on the suprarenal gland of the frog Rana temporaria in vivo and in vitro].

Biochemical and morphometric analyses have been made of the effects of nonapeptide neurohormones, vasotocin and mesotocin, on the chromaffin tissue of the adrenal gland of the frog, both in vivo and in vitro experiments. It was shown that vasotocin exerts stimulating effect on the suprarenal gland whereas mesotocin inhibits the latter. In vitro experiments revealed that adrenalin may abolish the effect of nonapeptides on the suprarenal gland.

Adrenal Glands↗