Evolutionary Advantages of Participation of Vasopressin instead of Vasotocin in Regulation of Water–Salt Balance in Mammals

In experiments on non-anesthetized Wistar white rats there was studied reaction of kidney to an intramuscular injection of arginine vasotocin or arginine vasopressin at doses from 0.001 to 0.05 µg/100 g body mass on the background of a water load. Water (5 ml/100 g body mass) was administered through a catheter into stomach to suppress secretion of endogenous antidiuretic hormone (ADH). In experiments with water administration, diuresis increased due to a decrease of osmotic permeability of renal tubules and to excretion of osmotically free water, with the constant clearance of sodium ions. Injection of 0.05 µg arginine vasopressin led to a marked decrease of diuresis due to a rise of reabsorption of osmotically free water without elevation of excretion of osmotically active substances. Injection of the same dose of arginine vasotocin resulted in no increase of diuresis; however, reabsorption of osmotically free water and excretion of osmotically active substances including sodium ions were more pronounced. Hence, both vasotocin and vasopressin increased osmotic permeability of the tubular epithelium, but vasotocin, unlike vasopressin, promoted reduction of reabsorption of sodium ions and their loss with urine. A suggestion is made that one of the reasons for replacement in mammals of the molecular ADH forms (vasotocin by vasopressin) was the absence of the pronounced natriuretic effect in arginine vasopressin. This was of crucial significance to preserve sodium ions in the organism, to maintain water–salt balance in animals adapted to the terrestrial life, and to provide not only osmo-, but also volumoregulation.     

Functional role of V1- and V2-receptors of the apical and basolateral membranes of the frog bladder epithelial cells

Arginine vasotocin, 0.02--1 nM, increases osmotic water permeability of frog urinary bladder, arginine vasotocin after a simultaneous addition to the mucosal and serosal Ringer solutions rises the water permeability to a lesser degree than on the hormone addition only to the serosal solution. 1 nM remestyp, an agonist of V1-receptors, from the apical membrane decreases the hydroosmotic effect of arginine vasotocin added to the serosal Ringer solution. When added to the mucosal solution, combination of the same concentrations of arginine vasotocin and SR 49059, an antagonist of V--receptors, or desmopressin, agonist of V2-receptor alone, increases the effect of the same concentration of arginine vasotocin added to the serosal solution. 1 nM arginine vasotocin at the luminal membrane increases secretion into the Ringer solution of prostaglandin E, and prostaglandin E1 but not of prostaglandin F2 alpha. The data obtained indicate the presence of the arginine vasotocin receptors responsible for the hydroosmotic effect only in the basolateral membranes, while arginine prostaglandin E, participation is shown in modulation of the arginine vasotocin effect.     

Renal filtration and reabsorption of GFP in frog Rana temporaria: effect of arginine-vasotocin

The renal tubular uptake of green fluorescent protein (GFP) in frog Rana temporaria was studied by laser confocal microscopy. The specific green fluorescence was revealed in the proximal tubule cells 30 min after intravenous GFP injection. The GFP fluorescence was distributed predominantly in the apical part of the cytoplasm in the form of the intensively fluorescing vesicles. The GFP injections increased dose-dependently the GFP tubular uptake. This was confirmed by the quantitative assessment of intensity of the specific fluorescence, its relative vesicular density, and by correlation analysis. Preliminary administration of arginine vasotocin into the dorsal lymphatic sac decreased significantly the GFP absorption. The effect of arginine vasotocin was inhibited by pretreatment a vasopressin V1-receptor antagonist. These results suggest that a decrease in the GFP absorption is due to a fall of the AVT-dependent glomerular filtration rate and consequently a decrease in the filtered GFP amount. The effect of arginine vasotocin on the GFP absorption seems to be mediated via the V1-like receptors of preglomerular blood vessels.     

Correlation of renal solute-free water reabsorption and energy of interaction of vasotocin analogs with V2 receptor

The nonapeptide-receptor interaction energy calculated from docking algorithms was compared with the effect of the same peptides upon solute-free water reabsorption in rat kidney in vivo. Vasotocin and some of its analogs were injected intramuscularly into conscious female Wistar rats in doses from 0.1 pmol to 0.5 nmol/kg of body weight after oral water load (50 ml/kg of body weight). A significant correlation was found between the calculated energy of peptide interaction with V₂ receptors and an increase in the renal solute-free water reabsorption stimulated by injection of nonapeptides. The results evidence that physiological changes in rat kidney in vivo induced by vasotocin analogs can be accurately simulated by virtual modeling of their interactions with V₂ receptors.     

Synthesis of new vasotocin analogues: effects on renal water and ion excretion in rats

Vasopressin and nonmammalian hormone vasotocin are known to increase the water permeability of mammalian collecting ducts, frog skin and the urinary bladder. Neurohypophysial nonapeptides have also been shown to interfere with the regulation of renal ion transport. The subject of this study was a search for vasopressin and vasotocin analogues with selective effects on renal water, sodium and potassium excretion. During this study, we synthesised the following peptides: 13 vasotocin analogues modified at positions 4 (Thr or Arg), 7 (Gly or Leu) and 8 (d ‐Arg, Lys or Glu); 4 vasopressin analogues modified at positions 4 and 8; and 9 peptides shortened or extended at the C‐terminal or with substitutions for Gly‐NH₂. Most of these peptides had mercaptopropionic acid (Mpa) instead of Cys in position 1. The effects of these nonapeptides on renal water, sodium and potassium transport were evaluated in in vivo experiments using Wistar rats. Some nonapeptides possessed antidiuretic, natriuretic and kaliuretic activities ([Mpa¹]‐arginine vasotocin, [Mpa¹, homoArg⁸]‐vasotocin, [Mpa¹, Thr⁴]‐arginine vasotocin and [Mpa¹, Arg⁴]‐arginine vasopressin). Substitutions at positions 4 and 8 increased the selectivity of peptide actions. The antidiuretic [d ‐Arg⁸]‐vasotocin analogues had no effects on sodium excretion. [Mpa¹, Arg⁴]‐arginine vasotocin was antidiuretic and kaliuretic but not natriuretic. [Mpa¹, Glu⁸]‐oxytocin had weak natriuretic activity without any effects on water and potassium transport. In accordance with the data obtained, synthesised vasotocin analogues could be good candidates for pharmaceuticals selectively regulating renal sodium and potassium transport, which is of clinical importance. Copyright © 2013 European Peptide Society and John Wiley & Sons, Ltd.     

Vasotocin acts as a dipsogen in ducks at concentrations stimulating subfornical organ neurons in vitro

The effects of systemic infusions of the avian antidiuretic hormone arginine vasotocin on water intake of domestic ducks were investigated under steady conditions of water balance in which angiotensin II was effective as a dipsogen. The study proceeded from the consistent stimulatory effect of arginine vasotocin on angiotensin II-responsive neurons found in the subfornical organ of ducks, suggesting brain-intrinsic vasotocinergic control of these neurons which are also accessible to circulating agents because of the lacking blood-brain barrier. Levels of circulating arginine vasotocin of about 2700 pg·ml^-1 which were close to the threshold for activation of subfornical organ neurons in vitro, induced weak but significant drinking responses. Even at this high arginine vasotocin level circulatory effects were absent, thereby excluding their interference with water intake. Arginine vasotocin plasma levels of about 60 pg·ml^-1 significantly attenuated the dipsogenic action of angiotensin. While drinking in response to high pharmacological levels of arginine vasotocin is assumed to mimic a stimulatory innervation of angiotensin-responsive subfornical organ neurons by brain-intrinsic vasotocinergic axons, attenuation of angiotensin-induced drinking by high physiological arginine vasotocin levels cannot be explained by its action on central neurons, but may be secondary to body fluid retention caused by the antidiuretic action of arginine vasotocin.Abbreviations ADH antidiuretic hormone - ANGII angiotensin II - AVP arginine vasopressin - AVT arginine vasotocin - BBB blood-brain barrier - HR heart rate - ICV intracerebroventricular - IV intravenous - MAP mean arterial pressure - SFO subfornical organ     

Plasma corticosterone of city and desert Curve-billed Thrashers, Toxostoma curvirostre, in response to stress-related peptide administration

We compared the activity and responsiveness of the hypothalamo-pituitary-adrenal (HPA) axis of an urban (Phoenix, Arizona) and desert population of a male songbird species (Curve-billed Thrasher, Toxostoma curvirostre), by measuring plasma corticosterone in response to acute administration of corticotropin-releasing factor, arginine vasotocin, or adrenocorticotropin hormone. Urban adult male thrashers showed greater responsiveness than desert birds to an injection of arginine vasotocin or adrenocorticotropin hormone, suggesting a population difference in pituitary and adrenal gland sensitivity. Plasma corticosterone in response to corticotropin-releasing factor injection did, however, not differ between populations. The differential corticosterone response to arginine vasotocin and corticotropin-releasing factor may reflect effects of chronic stress or habituation, which are known to favor arginine vasotocin over corticotropin-releasing factor sensitivity. Efficacy of HPA negative feedback by glucocorticoids was determined by measuring plasma corticosterone in response to acute administration of the synthetic glucocorticoid dexamethasone. This administration decreased plasma corticosterone similarly in urban and desert thrashers, suggesting that the negative feedback of glucocorticoids on the HPA axis in the two populations was equally effective. The higher sensitivity of urban than desert thrashers to adrenocorticotropin hormone and arginine vasotocin may result from up-regulation of the HPA axis in urban birds. This up-regulation may in turn make it easier for city birds to cope with urban environment-associated stressors.     

The effect of vasotocin on plasma glucose, free fatty acids and growth hormone in the frog, Rana catesbeiana

Injections of arginine vasotocin into bullfrogs had no effect on plasma glucose after 30 min. but caused hyperplycemia after 120 min. Plasma levels of growth hormone were elevated earlier (30 min.) and returned to control levels after 120 min. Plasma free fatty acids were unaltered following vasotocin administration. It is suggested that the hyperglycemic effect of vasotocin may be mediated via growth hormone.     

Testosterone response in arginine vasopressin desensitized immature rat testis

Direct injection of arginine vasopressin into immature rat testis inhibited basal testosterone synthesis. Simultaneous injection of arginine vasopressin with luteinizing hormone, norepinephrine or cholera toxin inhibited these agonists - induced testosterone response. In arginine vasopressin - desensitized testis, cAMP response to luteinizing hormone, norepinephrine and cholera toxin was not disturbed. However, testosterone response to luteinizing hormone, norepinephrine or cholera toxin was drastically reduced in arginine vasopressin-desensitized testis. This shows that the increased cAMP generated by luteinizing hormone, norepinephrine or cholera toxin in arginine vasopressin desensitized testis did not cause increase in steroidogenesis. This could be due to a lesion in steroidogenic pathway beyond cAMP generation caused by arginine vasopressin.     

Studies in vivo on the effects of parathyroid hormone upon kidney cyclic adenosine 3',5'-monophosphate content using rapid tissue fixation by microwave irradiation.

Microwave irradiation is shown to be a useful method for simultaneously killing chicks and fixing tissues. Renal adenylate cyclase and phosphodiesterase activities were rapidly abolished by microwaving. The increase in chick kidney cyclic adenosine 3',5'-monophosphate (cyclic AMP) content produced by intravenous bovine parathyroid hormone (PTH) injection was much greater in microwaved birds than in those killed by cervical dislocation with subsequent tissue fixation in liquid nitrogen. After PTH injection there was a prolonged elevation of renal cyclic AMP content. At the time of maximum response (2 minutes), log. dose-response curves were linear in the dose range 0.1-10 U. The responses to three different bovine PTH preparations were indistinguishable. Arginine vasopressin, arginine vasotocin, salmon calcitonin and prostaglandin E1 did not affect kidney cyclic AMP content within 2 minutes. Because of its specificity and precision, the method is of use for the in vivo bioassay of PTH. Injection of CaCl2 (20 mumoles) 1 minute before, or conjointly with, bovine PTH inhibited the subsequent increase in kidney cyclic AMP content. The synthetic bovine PTH peptide fragments BPTH (1-34) and BPTH (2-34) both increased chick kidney cyclic AMP content. The use of such fragments allows investigation of the structural requirements of PTH for interaction with the systems regulating cyclic AMP metabolism in the kidney in vivo.     

Central cardiovascular effects of mammalian neurohypophyseal peptides in conscious rats

To confirm and extend the results of previous studies which demonstrated central cardiovascular effects of vasopressin in anesthetized rats, we determined blood pressure and heart rate changes for 30 minutes after intracerebroventricular injections of arginine vasopressin, arginine vasotocin and oxytocin in conscious rats. As compared to sham injections, significantly greater increases in either systolic or diastolic blood pressure were noted over the 30 minutes which followed the injection of 0.15, 1.0 or 10.0 nM of either vasopressin or vasotocin. In animals given vasopressin, plasma levels of the peptide were determined. There was a substantial increase in plasma vasopressin only after the highest dose. Overall blood pressure responses to doses of oxytocin as high as 100 nM were not significantly different than sham injections. Heart rate following both vasopressin and vasotocin was increased at 0.15 nM, was initially decreased then increased at 1.0 nM and was substantially decreased after the 10.0 nM dose. There was a significant increase in heart rate at the 10.0 nM and 100 nM doses of oxytocin. Dose response curves for systolic blood pressure and heart rate 20 minutes after injection were similar for vasopressin and vasotocin. We conclude that arginine vasopressin has significant central pressor and tachycardic effects in conscious rats, and it is related, at least in part, to the tail structure of the peptide, which is shared with arginine vasotocin.     

Prolactin-releasing activity of arginine vasotocin in vitro.

A significant elevation in both luteinizing hormone (LH) and prolactin release was observed in the culture medium of hemipituitaries from castrated estrogen-progesterone (EP) primed female rats incubated for 5 h with arginine vasotocin (AVT) and luteinizing hormone-releasing hormone (LRH) compared to corresponding halves incubated with LRH alone. However, AVT alone did not significantly alter the discharge of LH or prolactin from hemipituitaries of EP-treated rats in vitro. Arginine vasopressin, a natural analogue of AVT, inhibited prolactin release using this same model system. Normal male rat hemipituitaries incubated with AVT released significantly more LH and prolactin into the culture medium than did their corresponding halves. A log-dose response curve indicated that any dose from 100 ng to 10 mug AVT significantly promoted prolactin release. However, the terminal tripeptide of AVT, Pro-Arg-Gly(NH2), failed to modify the discharge of LH or prolactin into the culture medium.     

Variation in the effects of antidiuretic hormone on the isolated skin of the toad, Scaphiopus couchi.

The rate of active sodium transport as measured by short-circuit current across the isolated skin of the toad, Scaphiopus couchi, was elevated following vasopressin (0.2 units/ml) or arginine vasotocin (0.1 units/ml) treatment of skins from active animals at all times of the year tested. Skins from dormant animals showed no such elevation at any time of the year. The rate of active sodium transport was elevated following treatment with dibutyryl cyclic AMP (2.5mM) plus theophylline (10 mM) in all skins tested. The hydraulic conductivity of isolated skins from both active and dormant animals showed no significant change following treatment with vasopressin (0.2 units/ml) or arginine vasotocin (0.1 units/ml except on the first day following emergence from dormancy in the field. A correlation was, therefore, observed between the occurrence of a hydroosmotic response to antidiuretic hormones and the seasonal exposure of S. couchi to standing water. A small but significant elevation of hydraulic conductivity was observed across the skins of dormant toads following treatment with dibutyryl cyclic AMP (2.5 mM) plus theophylline (10 mM) whereas a substantial elevation was observed with the skins of active animals.     

Immunocytochemical evidence for the presence of an albumin-like substance in the rat pineal gland

Summary Two rabbits and two guinea pigs were immunized with arginine vasotocin (AVT) conjugated to bovine albumin with glutaraldehyde. Only one preparation of antiserum (anti-G 7) was of value. Anti-G 7 immunochemically defined the same rat pineal cells previously reported as presumptive AVT cells. However, absorption of anti-G 7 with bovine albumin inhibited the staining of the pineal cells demonstrating that they contained an albumin-like substance. Positive immunochemical staining of the rat pars nervosa suggested that anti-G 7 contained antibodies able to react with arginine vasopressin (AVP). Loss of a positive reaction in the posterior lobe on absorption of anti-G 7 with AVT or AVP confirmed this. However, the addition of AVT to anti-G 7 failed to inhibit the immunochemical staining of the pineal cells. This study reports the presence of an albumin-like substance in pineal cells previously described as presumptive AVT cells, and discusses possible explanations for the inability of anti-G 7 to recognize immunocytochemically the native AVT molecule. Confirmation of AVT in the pineal gland by immunocytochemistry must await the availability of more specific antisera.     

Influence of ovine growth hormone on water and NaCl absorption by the rat proximal jejunum and distal ileum

1. Ovine growth hormone appears to stimulate equally the absorption of water and NaCl in both the proximal jejunum and in the distal ileum. 2. Although the largest dose used of 0.8 mg/rat for 2 days produced greater increases in absorption rates than the lowest dose of 0.1 mg, a clear dose-related response was not obtained. 3. It is suggested that growth hormone may contribute significantly in the regulation of hydromineral metabolism in mammals at times other than during pregnancy and lactation.     

Hormone-specific responses and biosynthesis of sulfolipids in cell lines derived from mammalian kidney.

The established cell lines isolated from mammalian kidney were characterized by its receptor activities against hormones and the ability to synthesize sulfolipids localized in the renal tubule. The level of 3':5'-cyclic AMP in JTC-12.P3 (monkey kidney) cells increased in 2 min as much as 2.5-5-fold on activation with 1.0 unit/ml of bovine parathyroid hormone or 1.9 units/ml of synthetic parathyroid hormone (1-34) resulting in intracellular cyclic AMP concentration of more than 40 pmol/mg protein. Prostaglandin E1 (14 micronM) and isopropylnorepinephrine (10 micronM) were also found to increase the concentration of cyclic AMP by more than 30- and 9-fold, respectively. Addition in medium of calcitonin, arginine vasopressin, adrenocorticotropic hormone and glucagon caused no significant changes of cyclic AMP level in the cell. In contrast, MDCK, a cell line isolated from canine kidney, reacted to arginine vasopressin, isopropylnorepinephrine and prostaglandin E1 and only slightly to parathyroid hormone. MDBK cell line derived from bovine kidney or fibroblast cell lines from rat lung and guinea pig kidney did not react to any of the hormones specific to kidney, i.e. arginine vasopressin, calcitonin or parathyroid hormone in the presence of theophylline. However, in the presence of 2 mM isobutylmethylxanthine, small but significant elevation of cellular cyclic AMP levels in response to calcitonin, arginine vasopressin, isopropylnorepinephrine and prostaglandin E1 was observed. The cell lines JTC-12, MDCK and MDBK, when incubated with H235SO4, incorporated the isotope into sulfolipids assigned as sulfatides and ceramide dihexoside sulfate or in MDCK also into cholesterol sulfate. The results suggested that JTC-12, MDCK and MDBK cell lines are epithelial origin and also JTC-12 and MDCK originated most probably from renal tubular cells of cortex and medulla, respectively.     

Sensory nerves contribute to insulin secretion by glucagon-like peptide-1 in mice

It has been hypothesized that the potent insulinotropic action of the gut incretin hormone glucagon-like peptide-1 (GLP-1) is exerted not only through a direct action on the beta cells but may be partially dependent on sensory nerves. We therefore examined the influence of GLP-1 in mice rendered sensory denervated by neonatal administration of capsaicin performed at days 2 and 5 (50 mg/kg). Control mice were given vehicle. Results show that at 10-16 wk of age in control mice, intravenous GLP-1 at 0.1 or 10 nmol/kg augmented the insulin response to intravenous glucose (1 g/kg) in association with improved glucose elimination. In contrast, in capsaicin-pretreated mice, GLP-1 at 0.1 nmol/kg could not augment the insulin response to intravenous glucose and no effect on glucose elimination was observed. Nevertheless, at the high dose of 10 nmol/kg, GLP-1 augmented the insulin response to glucose in capsaicin-pretreated mice as efficiently as in control mice. The insulin response to GLP-1 from isolated islets was not affected by neonatal capsaicin, and, furthermore, the in vivo insulin response to glucose was augmented whereas that to arginine was not affected by capsaicin. It is concluded that GLP-1-induced insulin secretion at a low dose in mice is dependent on intact sensory nerves and therefore indirectly mediated and that this distinguishes GLP-1 from other examined insulin secretagogues.     

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.     

Osmotic and volume control of diuresis in conscious ducks (Anas platyrhynchos)

Summary In conscious Pekin ducks made diuretic either by infusing hyposmotic glucose solution or isosmotic saline, osmotic and volume effects on renal water excretion were investigated. As in mammals, antidiuresis mediated by enhanced release of antidiuretic hormone was induced by increasing carotid blood osmolality while a decrease augmented diuresis, indicating cerebral osmotic control of renal water excretion in birds.In contrast to the situation in mammals, a sensitive diuretic response to isosmotic volume expansion, corresponding to 1% of the extracellular volume, can be demonstrated, with intracarotid and intravenous application of the isosmotic saline infusion having identical effects.Volume loading with isosmotic saline produced a greater diuretic response than loading with the same amount of autologous blood, thereby indicating a major contribution of volume changes in the interstitial compartment to the control of renal water excretion. This corresponds to the importance of the interstitial fluid compartment for the control of salt gland activity in this species.Abbreviations AVP arginine vasopressin - ECF extracellular fluid - i.c., i.v. intracarotid, intravenous - ECFV ECF volume     

Desensitization of immature rat testicular ornithine decarboxylase to arginine vasopressin

Prior exposure of immature rat testis to arginine vasopressin caused the testis refractory at 24 h in terms of ornithine decarboxylase activity. Arginine vasopressin caused desensitization both in Leydig cells and seminiferous tubules. Arginine vasopressin induced desensitization was found to be both time and dose-dependent. Arginine vasopressin desensitized testis was refractory to luteinizing hormone, follicle stimulating hormone, norepinephrine, dibutyryl cAMP, phorbol-myristate acetate and cholera toxin at 24 h. Arginine vasopressin desensitized testis showed recovery of response to norepinephrine at 48 h after the first injection. On the contrary arginine vasopressin could stimulate ornithine decarboxylase in luteinizing hormone desensitized testis. These results indicate that in arginine vasopressin desensitized testis the block is at post cAMP step which is common to both cAMP dependent and protein kinase C-diacylglycerol system in stimulating testicular ornithine decarboxylase.