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[Arg8]vasopressin-induced contractions of rabbit urinary bladder smooth muscle.

The effects of a number of neurohypophyseal hormones and analogues on the contractility of rabbit urinary bladder smooth muscle in vitro were examined. The order of potency was [Arg8]vasopressin = [Lys8]vasotocin greater than [Lys8]vasopressin greater than [Arg8]vasotocin much greater than [deamino1,D-Arg8]vasopressin greater than oxytocin much greater than pressinoic acid. The maximum tension induced by [Arg8]vasopressin was 4.4 N . cm-2 and the pD2 was 8.7. The effects of [Arg8]vasopressin were competitively antagonized by [(beta-mercapto-beta,beta-cyclopentamethylenepropionic acid)1,(O-methyl)Tyr2,Arg8]vasopressin and by [deaminopenicillamine1,Val4,D-Arg8]vasopressin with pA2s of 8.4 and 6.6, respectively. It was concluded that rabbit urinary bladder smooth muscle contains receptors for the neurohypophyseal hormones which have recognition properties similar to V1-vasopressin receptors.

Animals↗

Blood pressure homeostasis in the snake, Ptyas korros.

Three putative pressor systems, the alpha-adrenergic system (AS), the renin angiotensin system (RAS), and the arginine vasotocin system (AVT-S), were studied for their roles in blood pressure regulation and their possible interactions in the rat snake. Ptyas korros. Norepinephrine (NE), angiotensin I (ANG I), and arginine vasotocin (AVT) increased the mean arterial pressure (MAP) of the snake while administration of phentolamine, an alpha-adrenergic antagonist, and captopril, an ANG-converting enzyme inhibitor, but not KBIV24, an AVT antagonist, decreased the MAP. Treatment with any combination of two of these antagonists/inhibitor invariably decreased the MAP. Treatment with the agonist of the remaining third system invariably returned the MAP to normal or above. Phentolamine and KBIV24 attenuated the vasopressor effect of ANG I. Phentolamine and captopril enhanced the vasopressor effect of AVT. The pressor effect of NE was not altered by KBIV24 and captopril. It was concluded that there were at least two pressor systems (AS and RAS) regulating the basal MAP in the snake. There was also interaction among the three systems which could affect the MAP.

Angiotensin I↗

Direct control of the gonadotroph in a teleost, Poecilia latipinna. II. Neurohormones and neurotransmitters.

Pituitaries from male and female mollies were incubated with varying amounts of mammalian LH-RH, arginine vasotocin, dopamine, or serotonin for 18 hr. Ultrastructural differences between control and experimentally treated glands were used to define the direct effects of these neurohormones and neurotransmitters on the gonadotrophic cells of the adenohypophysis. The effects varied in intensity according to the sex and reproductive state of the donor animal. LH-RH stimulated gonadotrophin secretion by the gonadotrophs, as did vasotocin, although to a much lesser extent and with noticeable differences between the sexes. Dopamine inhibited secretion by basally active gonadotrophs and probably from active cells also, although to a lesser extent. Serotonin mildly stimulated secretion at all stages in both sexes. The results of this study indicate the possible involvement of neurohypophysial octapeptides and of monoamines in the direct control of the gonadotroph of Poecilia latipinna.

Animals↗

Sexual differences in hormonal control of release calls in bullfrogs.

Release calls in anuran amphibians are given when animals are inappropriately clasped by others. Since other call types, such as mate calls, are sexually dimorphic in frogs and toads, sonogram analysis was used to determine whether release call characteristics might also be sexually dimorphic in bullfrogs (Rana catesbeiana). Only intercall intervals differed significantly between males and females. Call duration, dominant frequency characteristics, and the display of secondary or tertiary harmonic frequencies were similar in both sexes. In the spring, but not the fall, calling rates were significantly lower in female bullfrogs, compared to males. Females also had significantly lower plasma androgen concentrations and higher plasma estrogen in the spring, compared to males. In both sexes, plasma androgen and estrogen were significantly higher in the spring, compared to the fall. The neuropeptide arginine vasotocin significantly decreased release call rates in females in the spring while it significantly increased rates in males. Vasotocin had no significant effect in the fall. Prostaglandin E2 significantly inhibited release calling in both males and females. On the other hand, prolactin significantly inhibited calling in female bullfrogs, but had no affect in males. Thus, although acoustic characteristics of release calls were similar in male and female bullfrogs, hormonal control of call rates was sexually dimorphic and seasonally variable.

Androgens↗

A seasonal pineal peptide rhythm persists in superior cervical ganglionectomized rats.

Neurohypophyseal peptide hormone activity is present in the pineal gland of mammals, and varies over a seasonal cycle. Pineal peptide levels, measured by arginine vasotocin (AVT) radioimmunoassay, increase dramatically for a brief time during August each year. The manner in which this cycle is regulated is as yet unknown. Input to the pineal from sympathetic axons arising in the superior cervical ganglia (SCG) is essential for the generation and regulation of the circadian rhythm in melatonin synthesis, and is the only pathway known to regulate pineal biochemical processes. It was of interest then to determine the impact of the SCG on the seasonal peptide cycle. Levels of pineal arginine vasotocin immunoactivity (iAVT) were monitored during August, 1984, in rats which had been superior cervical ganglionectomized (SCGX), in sham-operated and intact controls (L:D 12:12), and in rats subjected to L:D 22:2. The results indicate that SCGX does not abolish the seasonal cycle, but may influence the timing of the iAVT peak. Inhibition of pineal melatonin synthesis by exposure of rats to L:D 22:2 did not mimic the phase delay seen with SCGX, but did cause a significant increase in the amplitude of the August iAVT activity peak.

Animals↗

Regulation of oxytocin-induced phosphoinositide breakdown in adipocytes by adenosine, isoproterenol and insulin.

In rat adipocytes, the breakdown of phosphoinositides labelled by a 3 h incubation with [3H]inositol resulted in the accumulation of labelled inositol mono-, bis- and trisphosphates in the presence of oxytocin, vasotocin or vasopressin. Oxytocin at a concentration of 1 nM markedly increased phosphoinositide breakdown. Incubation of adipocytes both during the 3 h labelling and the 10 min breakdown period in a low adenosine medium (presence of adenosine deaminase) or high adenosine medium (presence of 0.1 microM N6-(phenylisopropyl)adenosine) (PIA) did not affect basal or ligand-stimulated phosphoinositide breakdown. The addition of 1 microM PIA only during the measurement of phosphoinositide breakdown variably stimulated basal breakdown but significantly potentiated that due to oxytocin. Isoproterenol similarly had little effect on basal but inhibited oxytocin stimulation of phosphoinositide breakdown. Insulin did not affect basal or ligand-stimulated phosphoinositide breakdown in the low or high adenosine medium. However, in adipocytes incubated in the absence of added adenosine deaminase or PIA, insulin stimulated basal accumulation of inositol phosphates by about 20% and inhibited that due to oxytocin by about 20%. There was no significant effect of insulin on the stimulation by vasopressin or vasotocin of phosphoinositide breakdown. These results indicate that, in adipocytes, phosphoinositide breakdown stimulated by oxytocin is enhanced by adenosine, inhibited by isoproterenol and, under some conditions is inhibited by insulin.

Adenosine↗

A new tritiated oxytocin receptor radioligand--synthesis and application for localization of central oxytocin receptors.

A new tritiated oxytocin antagonist radioligand was synthesized by introducing a tritiated propionic acid residue into the free amino group of ornithine in position 8 of the parent peptide [1-(beta-mercapto-beta,beta-cyclopentamethylene propionic acid), 2-(O-methyl)-tyrosine, 4-threonine, 8-ornithine, 9-tyrosylamide]vasotocin (OTA), that was previously described. The tritiated compound [3H][1-(beta-mercapto-beta,beta-cyclo-pentamethylene propionic acid), 2-(O-methyl)-tyrosine, 4-threonine, 8-(N6-propionyl)-ornithine, 9-tyrosylamide]vasotocin ([3H]PrOTA) was obtained in good yield with high specific activity (100 Ci/mmol). [3H]PrOTA exhibited the same affinity (Kd = 0.8 nM) and selectivity for the myometrial oxytocin receptor as the iodinated antagonist [125I]OTA. Autoradiographic localization of oxytocin receptors in the rat brain showed specific binding sites for [3H]PrOTA within regions of the limbic system, the neocortex, and hypothalamus, which is consistent with the binding pattern obtained with [125I]OTA. The high specific activity in combination with the long half-life of tritium and its low radiotoxicity as compared to iodine-125 makes the new tritiated antagonist a valuable tool for pharmacological studies.

Animals↗

Water balance of five amphibian species at different stages and phases, as affected by hypophysial hormones.

Five amphibian species were studied for the effect of hypophysial hormones on their water balance. The species were three anurans, Rana ridibunda, Bufo viridis and Pelobates syriacus, and two urodeles Salamandra salamandra and Triturus vittatus. In the first four species different stages of development were studied, in the newt both the terrestrial and aquatic phases of the adult were examined. The hormones used were oxytocin (OXY), arginine vasotocin (AVT) and prolactin (PL). Oxytocin caused most water retention when compared with the other hormones, especially responding were juveniles of Rana and Bufo, but also the terrestrial phase of the adult newt Triturus. Arginine vasotocin affected mostly juvenile Pelobates. Prolactin caused water retention in juvenile Rana and in the terrestrial phase of Triturus. In general the hormones affected the juvenile stages more than either larvae or adults.

Amphibians↗

L-cystine-di-beta-naphthylamidase activity in plasma of the snake Bothrops jararaca after chronic challenges of water balance.

Plasma protein, osmolality and L-cystine-di-beta-naphthylamidase activity were measured in Bothrops jararaca snakes submitted to normal hydration, water deprivation, water loading, salt loading or argvasotocin injections. Protein and osmolality were similar for male and female snakes in all treatments. Protein content was similar in all treatments, indicating the ability of the animals to maintain fluid balance between the vascular and extravascular compartment under osmotic challenges. Osmolality was increased in water-deprived and salt-loaded and was decreased in water-loaded animals compared with normally hydrated or argvasotocin-injected snakes. Enzymatic activity differed between males and females and was higher in salt-loaded females and in arg-vasotocin-injected females and males than in water-loaded snakes. A positive linear regression between osmolality and L-cystine-di-beta-naphthylamidase was obtained for female snakes (correlation coefficient = 0.6162, P < 0.01). The data show a role of arg-vasotocin in the consistent relationship between water balance and this sex-related aminopeptidase activity.

Aminopeptidases↗

Neurohypophysial hormones of dogfish, Triakis scyllium: structures and salinity-dependent secretion.

Sharks and rays utilize a unique strategy for adaptation to the hyperosmotic marine environment by maintaining their plasma slightly hyperosmotic to surrounding seawater (SW) through the accumulation of urea. Since neurohypophysial hormones (NHs) are plausible candidates for osmoregulatory effectors, the synthesis and release of NHs were investigated after transfer of fish to different environmental salinities. Molecular cloning revealed three NHs from the hypothalamus of a dogfish, Triakis scyllium: vasotocin (VT), asvatocin, and a novel oxytocin-family peptide, phasitocin ([Phe3, Asn4, Ile8]vasotocin). The VT precursor consists of a signal peptide, VT, a neurophysin and a copeptin moiety. In contrast, the asvatocin and phasitocin precursors are shorter due to the lack of a copeptin moiety as is the case in oxytocin and mesotocin precursors in tetrapods and lungfish, but different from teleost isotocin precursors that have the copeptin moiety. In the hypothalamus, VT mRNA levels significantly increased after transfer to concentrated (130%) SW for 2 days, while no change was observed in mRNA levels of asvatocin and phasitocin following transfer to either 130% or diluted (60%) SW. The increase in VT mRNA was reflected in the plasma level of peptide; plasma VT concentration measured by highly sensitive and specific radioimmunoassay increased according to elevated environmental salinities. These results suggest that VT is an osmoregulatory effector in dogfish, especially when the dogfish is exposed to a hyperosmotic environment.

Amino Acid Sequence↗

FSH and LH-beta subunits in the preoptic nucleus: ontogenic expression in teleost.

In the present study we cloned, sequenced, and confirmed the presence of mRNAs of gonadotropins (FSH-beta, LH-beta subunits) from the brain and pituitary of tilapia, Oreochromis niloticus. Further, we examined the spatio-temporal expression pattern of FSH-beta and LH-beta in the brain and pituitary of two species of teleost (tilapia, O. niloticus; sockeye salmon, Oncorhynchus nerka), using in situ hybridization and immunological methods. The expression of FSH and LH immunoreactivity appeared simultaneously in the brain and pituitary (tilapia, 14 days; sockeye, 51 days after fertilization). In the pituitary, FSH mRNA and peptide expressing cells were distinct from LH expressing cells located in the ventral proximal pars distalis. In the brain, FSH and LH immunoreactivity was co-localized in cells of the preoptic nucleus parvocellularis, magnocellularis, and gigantocellularis. Fibers immunoreactive to FSH and LH antisera were seen along the forebrain-hypothalamus and in the neurohypophysis of the pituitary. Double-label immunofluorescence revealed FSH and LH immunoreactivity co-localized in arginine vasotocin synthesizing preoptic neurons. Our results show that FSH and LH-producing cells have developmental origins in the brain as well as in the pituitary. In addition, we propose that the brain-derived gonadotropins may function as hypophysiotropic hormones that regulate pituitary cells and along with arginine vasotocin could act as neuromodulators of reproductive behaviors.

Age Factors↗

Oxytocinergic neurotransmission at the hippocampus in the central neural regulation of penile erection in the rat.

OBJECTIVES: To investigate whether there is an oxytocinergic neurotransmission at the hippocampus involved in the central regulation of penile erection in the rat. METHODS: Male adult Sprague-Dawley (n = 27) rats anesthetized with pentobarbital were used. A 26-gauge needle was inserted into the corpus cavernosum to measure the intracavernous pressure (ICP) and the systemic and mean arterial pressure and heart rate simultaneously. The following studies were performed: stereotaxic delivery of oxytocin acetate (3 pmol/100 nL) into the hippocampus; microinjection of a mixture of [d(CH(2))(5)-Tyr(Me)(2)-Orn(8)]-vasotocin (3 pmol/100 nL) and oxytocin (3 pmol/100 nL) into the hippocampus; injection of saline into the hippocampus; and intracavernous injection of oxytocin (3 pmol/100 microL). The ICP and hemodynamic parameters were monitored after each administration of the experimental agents. RESULTS: After administration of oxytocin into the hippocampus, a significant increase in the ICP occurred from resting (8.8 +/- 1.7 mm Hg) to a peak at 49.6 +/- 12.5 mm Hg and persisted for 18.6 +/- 9.4 minutes after an onset latency of 500.0 +/- 389.7 seconds. However, no change in the ICP occurred after administration of [d(CH(2))(5)-Tyr(Me)(2)-Orn(8)]-vasotocin plus oxytocin into the hippocampus. In addition, no elevation of ICP occurred after administration of saline to the hippocampus or after intracavernous injection of oxytocin. CONCLUSIONS: The results demonstrate that administration of oxytocin into the hippocampus induces penile erection in the rat. However, concomitant administration of oxytocin and its antagonist was ineffective in eliciting penile erection. These observations suggest that oxytocinergic neurotransmission at the hippocampus may be involved in the central neural regulation of the penile erection in the rat.

Animals↗

Neuroendocrine regulation of GnRH and behavior during aging in birds.

Avian species exhibit a great variety of life-long patterns in reproduction. Japanese quail are relatively short lived and undergo an age-related loss of reproductive function, making this species an excellent model for the study of the basic biology of aging. Because individuals age at variable rates, sexual behavior has provided a useful index to assess reproductive status of individuals of the same chronological age. Further, exogenous testosterone restores sexual behavior in reproductively senescent male quail, thereby providing evidence for a continued ability of the system to respond. In addition, we have been studying hypothalamic neuroendocrine systems that regulate the endocrine as well as behavioral components of reproduction. Overall, our findings point to the hypothalamic neuroendocrine systems as the site of initial age-related alterations that contribute to the reproductive deterioration. Specifically, we studied adrenergic, opioid peptide, vasotocin, and aromatase systems to understand their relationship to the cGnRH-I system and their potential role in the deterioration of the cGnRH-I system during aging. Our findings provide evidence for qualitative and quantitative alterations in the aromatase enzyme system, which can be partially restored with exogenous testosterone. In addition, other neuronal systems, including the vasotocin system, decline with the loss of gonadal steroids and are restimulated with exogenous testosterone. We will synthesize the data relative to these neuroendocrine systems with attention to the effects of gonadal steroids on these systems during aging.

Aging↗

Combined use of immunocytochemistry and lectin histochemistry for the study of the hypothalamic neurosecretory system of the snake Natrix maura (L.).

Natrix maura snakes were processed for immunocytochemistry and lectin histochemistry at both light- and electron-microscopic levels. Antisera against bovine neurophysins, vasotocin and mesotocin were used as well as concanavalin A, wheat germ and Limax flavus agglutinin lectins. The hypothalamic supraoptic and paraventricular nuclei were studied. Vasotocin neurons should contain a glycopeptide and displayed large colloid droplets consisting of large cisternae filled with packed secretory material. Mesotocin was located in different neurons.

Animals↗

Prostaglandin-dependent osmotic water permeability of the frog and trout urinary bladder.

Washout of autacoids from serosal Ringer solution, using a repeated change of the solution of the frog and trout urinary bladder, was accompanied by a pronounced rise in the osmotic water permeability: the water transport in the frog rose from 0.05 +/- 0.02 to 1.21 +/- 0.26 microliter min-1.cm-2, in the trout, from 0.041 +/- 0.011 to 0.26 +/- 0.034 microliter min-1.cm-2. Such an increase in the osmotic water permeability in the trout and frog urinary bladder occurred in the background of a decrease in the prostaglandin E2 concentration in the serosal Ringer solution. This permeability increase was accompanied by the formation of aggregates of intramembranous particles in the apical plasma membrane of the trout and frog urinary bladder. A decrease in the osmotic water permeability was achieved by the addition to the serosal Ringer solution of 10-8 M prostaglandin. Experiments on the frog urinary bladder have shown that prostaglandins E1, I2 and F2 alpha also decrease the osmotic water permeability. Vasotocin increased the osmotic water permeability in the frog urinary bladder but did not affect the osmotic water permeability of the trout urinary bladder. The data obtained indicates a role of the endogenous prostaglandin production in maintaining the low osmotic water permeability in the frog and trout urinary bladder. A suggestion is made that in the vertebrate evolution, colonisation of the fresh-water was connected with the maintenance of the low osmotic water permeability via participation of prostaglandins, whereas the vasotocin hydroosmotic effect developed in the vertebrate evolution later and provided for the possibility of the water absorption, osmotic homeostasis and animal migration from fresh-water to the land.

Alprostadil↗

Possible absorption of antidiuretic hormone in isolated rat intestine.

Arginine-vasopressin or arginine-vasotocin hormones introduced into the small intestine are absorbed and exert a hydrosmotic effect. The amount of absorbed antidiuretic hormone increased when it is introduced with intestinal peptidase inhibitor aprotinin. These data attest to the possibility of absorption of pituitary hormones (vasotocin and vasopressin) in rat small intestine without losing their physiological activity.

Animals↗

Hormonal response of primary hepatocytes of the clawed toad, Xenopus laevis.

Freshly isolated hepatocytes of the clawed toad, Xenopus laevis, were cultured for at least 3 days. The viability of the cells was characterized using staining and biochemical methods. In particular, the glucose and glycogen balance was tested. After culture for 16-20 hrs, the cells were subjected to hormonal treatment. Both adrenaline and arginine vasotocin stimulated the release of glucose in a dose dependent manner. 10(-6) M concentrations were strongly effective. The determination of the glycogen balance made it clear that the glucose release is mainly due to glycogenolysis. Using receptor antagonists and agonists, it has been shown that the effect of adrenaline is clearly mediated by beta-type receptors. Arginine vasotocin stimulated glycogenolysis via a type of receptor which is similar to the V2-receptor of mammals. This means that cAMP is involved in the response to both types of hormones which is in contrast to that which is known about the effect of nonapeptides on the liver of mammals.

Animals↗

Ventilation, sensible heat loss, broiler energy, and water balance under harsh environmental conditions.

Air velocity (AV) is one of the main environmental factors involved in thermoregulation, especially at high ambient temperatures. To elucidate the effect of AV on performance and thermoregulation of 4- to 7-wk-old broiler chickens, an experiment was conducted using 4 different AV (0.8, 1.5, 2.0, and 3.0 m/s) at constant ambient temperatature (35 +/- 1.0 degrees C) and RH (60 +/- 2.5%). BW, feed intake, and fecal and urinary excretions were monitored in individuals and were used to calculate the amount of energy expended for maintenance. Infrared thermal imaging radiometry was used to measure surface temperatures for the calculation of heat loss by radiation and convection. Brachial vein blood was collected for plasma osmolality and arginine vasotocin analysis. Broilers performed optimally at an AV of 2.0 m/s. Energy expenditure for maintenance was significantly higher under these conditions, suggesting the ability to direct a sufficient amount of energy to control body temperature, while maintaining relatively high growth rates. Convective heat loss increased significantly with increasing AV, whereas radiative heat loss was not affected. Sensible heat loss, expressed as a percentage of energy expenditure for maintenance, was significantly higher at 2.0 m/s compared with 0.8 m/s but significantly lower than that of 3.0 m/s. The high level of heat loss observed at 3.0 m/s probably affected body water balance, as supported by significantly higher plasma osmolality, arginine vasotocin concentration, and the hyperthermic status of these birds. It can be concluded that AV of 2.0 m/s enables broilers to maintain proper performance together with efficient thermoregulation and water balance under harsh environmental conditions.

Air Movements↗