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11-Ketotestosterone inhibits the alternative mating tactic in sneaker males of the peacock blenny, Salaria pavo.

In the peacock blenny, Salaria pavo, a species with courtship sex-role reversal, smaller, younger males mimic the courtship behavior and the nuptial coloration of females in order to get access to nests during spawning and to parasitize egg fertilization from nest-holder males. Later in their life, sneakers transform both morphologically and behaviorally into nest-holder males. In the present paper we investigate the activational role of 11-ketotestosterone (KT), the most potent androgen in most teleost species, to promote the switch between tactics in sneaker males of S. pavo. Sneakers were implanted either with KT or with control (i.e. castor oil) silastic implants. A week after implantation they were subjected to a set of behavioral tests and morphometric measurements. KT treatment promoted the differentiation of secondary sex characters, such as the anal glands, and inhibited the expression of female courtship behavior. KT-treated sneakers also showed a trend toward less frequent display of female nuptial coloration. There was no effect of KT treatment on the expression of typical nest-holder male behavior. Finally, there was no effect of KT treatment on the number or soma size of arginine vasotocin neurons in the preoptic area, which are often associated with the expression of vertebrate sexual behavior. Thus, KT seems to play a key role in mating tactic switching by inhibiting the expression of female courtship behavior and by promoting the development of male displaying traits (e.g. anal glands). The lack of a KT effect on behavior typical of nest-holding males and vasotocinergic preoptic neurons suggests that a longer time frame or other endocrine/social signals are needed for the initiation of these traits in males that are switching tactics.

Animals↗

Expression of nitric oxide synthase in the preoptic-hypothalamo-hypophyseal system of the teleost Oreochromis niloticus.

In the present study, we have analyzed the expression of nitric oxide synthase (NOS) in the preoptic-hypothalamo-hypophyseal system of the teleost Oreochromis niloticus. The assay for enzyme activity demonstrated that a constitutive NOS activity is present both in soluble and particulate fractions of the homogenates of diencephalons. Western blot analysis using an antibody against the N-terminus of human nNOS revealed two bands both in the supernatant and in the pellet. One band co-migrates at approximately 150 kDa with that detected in the rat cerebellum homogenates and presumably corresponds to neuronal NOS (nNOS) of mammals. The additional band, which migrates at approximately 180 kDa, might be attributed to an alternatively spliced nNOS isoform. Using NADPH diaphorase (NADPHd) histochemistry in combination with NOS immunohistochemistry, nNOS expression has been detected in preoptic nuclei, hypophysiotrophic nuclei of the ventral hypothalamus, and the pituitary gland. Various degrees of dissociation of NADPHd activity and nNOS immunoreactivity have been detected that could be attributed to the expression of different subtypes of nNOS in the preoptic/hypothalamo/hypophysial system of tilapia. In this paper, we also investigated the colocalization of nNOS with arginine-vasotocin (AVT) by means of immunolabeling of consecutive sections. Results suggest that NO may be colocalized with AVT in a subpopulation of neurosecretory neurons. Present findings suggest that nitric oxide (NO) is implicated in the modulation of hormone release in teleosts in a similar way to mammals.

Animals↗

Radioimmunoassayable avt and avp in adult mammalian brain tissue: comparison of normal and brattleboro rats.

Arginine vasopressin (AVP) and vasotocin (AVT) were measured by radioimmunoassay in extracts of cerebral cortex, cerebellum, brain stem, pineal, hypothalamus, and neurohypophysis from normal Long-Evans rats. AVP was present in expected concentrations in pituitary and hypothalamus and there was no evidence of its accumulation elsewhere. AVT was not detectable in these tissues (within the limits imposed on our assay by the presence of excessive amounts of AVP) but was easily detectable in pineal tissue with a concentration of 22.4 plus or minus 6.6 muU/gland. Extracts of neurohypophysis and pineal glands from homozygous Brattleboro rats (rats with hereditary hypothalamic diabetes insipidus) revealed the total absence of AVP and AVT. We conclude that the Brattleboro rat is incapabel of synthesizing biologically active neurohypophyseal peptides which contain arginine in position 8.

Animals↗

Occurrence of hydrin 2 (vasotocinyl-Gly), a new hydroosmotic neurohypophyseal peptide, in secretory granules isolated from the frog (Rana esculenta) neurointermediate pituitary.

Neurohypophyseal secretory granules have been purified from the frog (Rana esculenta) neurointermediate pituitary gland by sucrose gradient centrifugation, and their polypeptide content has been analyzed by reverse-phase high-pressure liquid chromatography. Aside from vasotocin, mesotocin, and their associated neurophysins, hydrin 2 (vasotocinyl-Gly), previously identified in hydrochloric acid extracts, has been recognized. This finding supports the previous suggestion that hydrin 2, a peptide active on the water permeability of frog bladder and frog skin, is a secreted hormone involved in osmoregulation specific to amphibians. Hydrin 2 has not been found in neurosecretory granules of birds such as the goose.

Animals↗

Studies on the role of the pineal gland in the regulation of TSH secretion in postpuberal male rats.

The role of the pineal gland in the regulation of thyroid-stimulating hormone (TSH) secretion was studied in postpuberal male rats using 12:12 h light:dark cycles. Pinealectomy, performed 3--4 days before decapitation, had no effect on the basal or thyrotropin-releasing hormone stimulated TSH secretion, but significantly decreased the cold-induced TSH response both at noon and at midnight. Neither hypothalamic thyrotropin-releasing hormone concentrations nor serum thyroid hormone levels were affected by pinealectomy. Melatonin injected intravenously (1 and 10 mg/kg) or intracerebroventricularly (5 and 50 microgram/rat) had no effect on the basal or cold-stimulated TSH levels, neither was the thyrotropin-releasing hormone induced (50 and 125 ng/rat i.p.) TSH response modified by melatonin (1 mg/kg i.v.). Arginine vasotocin (1 and 100 ng/rat) given intracerebroventricularly had no effect on the basal or cold-stimulated TSH secretion. These results suggest that the pineal has no influence on the basal activity of the hypothalamus-pituitary-thyroid system, but it seems that the cold-induced TSH response depends to a considerable degree on the intact pineal gland.

Animals↗

Hormonal regulation of renal function during development.

The present review summarizes current and new knowledge concerning the major hormonal systems that directly or indirectly affect renal function during development. The role of the renin-angiotensin-aldosterone system in regulating renal function during fetal and postnatal life is reviewed. A summary of the role of this system during fetal and postnatal stresses is also provided. The physiological role of the renal kallikrein-kinin system in the control of renal blood flow, renin release and sodium excretion during development is examined. Possible influences of the prostaglandin system on regulation of renal function and renin secretion during fetal and postnatal maturation are explored. The effect of vasopressin on the ability of the fetal and postnatal kidney to concentrate urine and regulate body fluid homeostasis is reviewed in detail. The physiologic action of vasotocin on renal sodium and water homeostasis is described. New information regarding the role of the sympathetic system in the regulation of renal hemodynamics and in the control of renal function during development is presented. Finally, recent studies demonstrating the effect of atrial natriuretic factor and corticosteroids on the developing kidney are discussed.

Animals↗

Glomerular effects of AVT on the in situ perfused trunk preparation of the dogfish.

The effects of arginine vasotocin on the pattern of glomerular perfusion in an elasmobranch fish were examined using an in situ perfused renal trunk preparation. A significant antidiuresis was coupled with a marked reduction in the proportion of filtering glomeruli (86%-25%) and an increase in the proportion of perfused but not filtering (10%-53%) and non-perfused glomeruli (4%-22%). However, the reduction in filtering glomeruli was greater than predicted by measurements of urine flow rate and glomerular filtration rate alone, suggesting that alterations in single nephron glomerular filtration rate may occur.

Animals↗

Cortisol facilitates ovine fetal/maternal water transfer.

After the infusion of 500 ml 20% mannitol to 12 pregnant ewes, we studied fetal plasma osmolality responses in chronically catheterized fetal lambs at 129-140 days' gestation. To compare the effects of arginine vasotocin (AVT) alone and AVT plus cortisol on fetal/maternal water transfer, the mannitol infusion was repeated during a 2-h fetal infusion of AVT alone (7 microU/kg/min) and during infusion of AVT with cortisol (50 micrograms/h) (eight and six animals, respectively). The fetal osmolar response to maternal mannitol administration was compared to the response following the baseline mannitol study in the same sheep. In four of the ewes, an identical (sham) study was performed substituting 500 ml normal saline for the mannitol infusion to the ewe. Fetal AVT infusion significantly obtunded the fetal osmolality increment induced by maternal mannitol alone [P less than 0.001, analysis of variance (ANOVA)]. Fetal AVT administered concurrently with cortisol produced an increment in fetal plasma osmolality in response to maternal mannitol that was significantly enhanced compared with that during fetal AVT infusion alone (P less than 0.001, ANOVA). The response to AVT with cortisol was similar to that following mannitol alone. Normal saline (sham protocol) produced no change in maternal or fetal osmolality. These results indicate that fetal AVT inhibits fetal to maternal water transfer following an osmotic stimulus to the ewe, while cortisol (infused with AVT) tends to counteract the inhibitory effect of AVT on fetal/maternal water transfer.

Analysis of Variance↗

Female oxytocin-deficient mice display enhanced anxiety-related behavior.

Previous studies have suggested that oxytocin (OT) may be anxiolytic in female laboratory rats and mice. The elevated plus-maze was used to compare anxiety-related behaviors of OT-deficient (OT-/-) and wild-type (OT+/+) mice. Female OT-/- mice displayed increased anxiety-related behavior compared with OT+/+ mice. The percentage of entries (P < 0.0002) and time spent (P < 0.003) in the open arms was less in female OT-/- than OT+/+ mice. Administration of synthetic OT, 2 ng by intracerebroventricular (icv) injection to female OT-/- mice, increased the percentage of entries (P < 0.003) and time spent (P < 0.004) in the open arms compared with artificial cerebrospinal fluid female OT-/- mice. Administration of an OT receptor antagonist (Atosiban, d[Dtyr(Et)(2), Thr(4)]ornithine vasotocin) 100 ng icv, to female OT+/+ mice increased anxiety-related behavior by decreasing the percentage of entries (P < 0.01) and time spent (P < 0.04) in the open arms compared with artificial cerebrospinal fluid-treated controls. Central infusion of an OT receptor antagonist, 100 ng icv, before administration of synthetic OT, 2 ng icv, in female OT-/- mice blocked the anxiolytic affect of OT. In contrast, male OT-/- mice displayed decreased anxiety-related behavior compared with male OT+/+ mice. The percentage of entries (P < 0.007) and time spent (P < 0.004) in the open arms was greater in male OT-/- vs. OT+/+ mice. Our findings indicate that OT pathways play a role in modulating anxiety in female mice of the C57BL/6 background, and the effect is mediated by the OT receptor.

Animals↗

Radioimmunologic detection and measurement of nonapeptides in the pineal gland.

RIAs have been developed for the nonapeptide hormones arginine vasotocin (AVT), arginine vasopressin (AVP), and oxytocin (OT). The AVP RIA can detect as little as 2 pg hormone and shows essentially no cross-reactivity with AVT or OT. The OT RIA is sensitive to 7 pg and shows no significant cross-reactivity with AVP or AVT. The AVT RIA is sensitive to about 5 pg; some cross-reactivity occurs with OT and AVP, but the RIA is suitable for assaying AVT levels in biological samples containing OT and/or AVP in concentrations up to 5 times greater than that of AVT. Using these RIAs, we found large amounts of AVT (up to 1.48 microgram/gland) in the chicken pituitary but no AVP or OT. The chicken pineal also contained AVT (about 300 pg/gland) and lacked AVP and OT. Bovine pineal glands appeared to contain all three peptides in roughly similar amounts (200-400 pg/gland). Pineal glands from a variety of rodents (including the rat) contained only very small amounts of AVT-like immunoreactivity (about 10 pg/gland) and no AVP or OT. Because AVT immunoreactivity appears in the pineals of several species, the peptide may subserve some physiological function of this organ. The functional roles, if any, of AVP and OT in the bovine pineal are unknown.

Animals↗

"Antigonadal" activity of the neurohypophysial hormones in cultured rat testicular cells: abolition by pertussis toxin.

The intermediary role of the putative inhibitory regulatory membrane protein "Ni" in the "antigonadal" activity of the neurohypophysial hormones was investigated in vitro with the use of a primary culture of rat testicular cells. To this end, use was made of the pertussis toxin (PT) probe, an exotoxin of Bordetella pertussis presumed to modify and inactivate a membrane protein related to or identical with Ni. Testicular cells were pretreated with PT (70 ng/ml) for 24 h, a duration of exposure known to result in a maximal PT effect. Thereafter, the cells were cultured for an additional 48 h in the absence or presence of hCG (10 ng/ml), with or without a maximal inhibitory dose (10(-6) M) of the neurohypophysial principle arginine vasotocin (AVT). Although concomitant treatment of control cells with AVT produced a profound (95%) inhibition of hCG-stimulated testosterone accumulation, PT pretreatment resulted in a long-lasting (greater than 4 days) abolition of this "antigonadal" effect. Furthermore, pretreatment with PT resulted in significant (P less than 0.05) augmentation of the hCG-stimulated accumulation of extracellular cAMP (1.5- to 2.0-fold) and testosterone (1.7- to 3.8-fold), presumably as a result of abolition of the "basal" tonic inhibition of adenylate cyclase activity. Taken together, these findings constitute the first demonstration of the involvement of Ni in the "antigonadal" activity of the neurohypophysial hormones and in the regulation of testicular function, thereby raising the intriguing possibility that testicular Ni may serve as the coupling unit of inhibitory receptors other than those for the neurohypophysial hormones, integrating varied incoming signals of endocrine, paracrine or autocrine nature.

Adenylate Cyclase Toxin↗

A novel type of vasopressin receptor on anterior pituitary corticotrophs?

Suspensions of rat anterior pituitary cells were exposed to corticotropin-releasing factor (CRF) (5 nM) and various neurohormones (0.002-1000 nM). CRF-induced secretion of ACTH was doubled by 0.1 nM arginine vasopressin (AVP), 0.2 nM arginine vasotocin, 1 nM oxytocin, 10 nM angiotensin II, and 100 nM noradrenalin; vasoactive intestinal peptide had no effect at 0.2-200 nM. CRF potentiation by AVP was also observed at lower concentrations of CRF. Since AVP appeared to be the most potent modulator of CRF-induced ACTH secretion, potentiation was further tested with specific antidiuretic and oxytocic agonists. Potentiation was clearly related to pressor biological activity, less so to antidiuretic, and hardly at all to oxytocic activities. However, even at 200 nM, the antipressor antagonists dPTyr(Me)AVP and d(CH2)5Tyr(Me)AVP had no effect on potentiation by AVP. The lack of antagonism was partly due to the agonistic effects of the antagonists on the pituitary gland, an effect not observed within vascular tissue. The results thus suggest that anterior pituitary vasopressin receptors resemble, but are not identical to, V1 (pressor and hepatic), do not resemble the V2 (renal), and might be classified as V3 (pituitary) receptors.

Adrenocorticotropic Hormone↗

Prolactin-releasing and release-inhibiting factor activities in the bovine, rat, and human pineal gland: in vitro and in vivo studies.

The effects of crude extracts of bovine, rat, and human pineal glands on prolactin (PRL) release were studied using an in vitro system. In addition, the effects of a known pineal constituent, arginine vasotocin (AVT), and crude bovine pineal extract (bPE) on PRL secretion were studied in vivo. Normal male rat hemipituitaries (HP), incubated with bPE (13 mg tissue/HP)released 200%, 150%, and 285% more PRL into the medium than did their corresponding untreated control halves incubated in either Medium 199 alone, hypothalamic extract, or cerebral cortical extract, respectively. HP incubated with either rat (6 mg of tissue/HP) or human (25 mg of tissue/HP) pineal extract released 110% and 75% more PRL, respectively, than did their corresponding untreated control halves. HP exposed to 10 mg tissue eq of either bovine pineal fraction A1 or bovine pineal fraction A3 released 88% and 63%, respectively, less PRL than did their corresponding untreated control halves incubated in Krebs-Ringer Bicarbonate (KRB) medium. Quantitites of melatonin, thyrotropin-releasing hormone (TRH), or estrogen, comparable to those found in the pineal, had no significant effect on PRL secretion in vitro. The iv injection of either bPE (90 mg tissue/rat) or AVT (10 mug/rat) into estrogen and progesterone-treated male rats resulted in a 40% and 138% increase, respectively, in plasma PRL titers, 10 min after injection, over pre-injection control levels. The per cent of increase in plasma PRL levels in these animals was significantly greater than that observed in control rats receiving either saline or cortical extract. The results suggest that crude extracts of pineal glands of three different species contain prolactin-releasing factor (PRF) activity which is probably not due to any endogenous melatonin, TRH, or estrogen that may be present. Conversely, two bovine pineal fractions, A1 and A3, appeared to exhibit prolactin-inhibiting factor (PIF) activity. We have concluded that the pineal gland may serve as an alternate or supplemental source of PRF and/or PIF.

Aged↗

Regulation of oxytocin receptor messenger ribonucleic acid in the ventromedial hypothalamus by testosterone and its metabolites.

Oxytocin receptor (OR) binding in the ventromedial hypothalamus (VMH) is regulated by testosterone (T) and its metabolites, estrogen (E2) and dihydrotestosterone (DHT). Previous studies have reported that OR binding increases in the VMH in castrated male rats when they are replaced with T or E2 compared to that in vehicle-treated animals. DHT alone had no effect on OR binding, but when given in combination with E2 appeared to have a synergistic effect. This study was designed to determine whether these effects of steroid hormones on OR binding in the VMH are associated with changes in OR messenger RNA (mRNA) expression. Male rats were castrated or sham operated and given T propionate (TP), E2 benzoate (EB), DHT plus EB, or an oil vehicle. OR mRNA was assessed using a rat complementary RNA OR probe and in situ hybridization techniques. OR binding to tissue slices was quantified autoradiographically using an OR antagonist, [125I]d(CH2)5[Tyr(Me)2,Thr4,Tyr-NH2(9)] ornithine vasotocin. These experiments showed that TP and EB increased both OR mRNA and OR binding in the VMH significantly above levels in vehicle-treated animals. However, animals given both EB and DHT exhibited significantly lower OR mRNA expression and OR binding in the VMH compared to those in animals treated with TP or EB alone. These data indicate that increases in VMH OR binding in response to gonadal steroids are accompanied by changes at the mRNA level that correspond well in magnitude and direction with those in the OR-binding sites.

Animals↗

Central effects of various ligands on drinking behavior in eels acclimated to seawater.

Intracranial injection of eel angiotensin II (eANG II, 5x10(-13)-5x10(-8) mol), acetylcholine (ACh, 5x10(-12)-5x10(-9) mol), substance P (5x10(-10) mol) and isoproterenol (a beta-adrenoceptor agonist, 5x10(-11)-5x10(-9) mol) enhanced water intake in the seawater eel. The effects of eANG II, ACh and isoproterenol were dose-dependent. By contrast, water intake was inhibited by intracranial injection of eel atrial natriuretic peptide (eANP, 5x10(-13)-5x10(-10) mol), serotonin (5-HT, 5x10(-12)-5x10(-8) mol), ghrelin (5x10(-12)-5x10(-10) mol), gamma-amino butyric acid (GABA, 5x10(-11)-5x10(-8) mol), prolactin (PRL, 5x10(-10)-5x10(-9) mol), arginine vasotocin (AVT, 5x10(-12) mol), vasoactive intestinal peptide (VIP, 5x10(-11) mol), noradrenaline (5x10(-9) mol l(-1)) and phenylephrine (alpha-adrenoceptor agonist, 5x10(-11)-5x10(-9) mol). The inhibitory effects of eANP, 5-HT, ghrelin, GABA, PRL and phenylephrine were dose-dependent. The intracranial stimulatory effect of eANG II was relatively long-lasting compared with the intravenous effect. The stimulatory effect of intravenous eANG II disappeared immediately, and was followed by an inhibition, which could be well explained by an increase in eANP secretion from the atrium.

Acclimatization↗

Water balance of field-excavated aestivating Australian desert frogs, the cocoon-forming Neobatrachus aquilonius and the non-cocooning Notaden nichollsi (Amphibia: Myobatrachidae).

Burrowed aestivating frogs of the cocoon-forming species Neobatrachus aquilonius and the non-cocooning species Notaden nichollsi were excavated in the Gibson Desert of central Australia. Their hydration state (osmotic pressure of the plasma and urine) was compared to the moisture content and water potential of the surrounding soil. The non-cocooning N. nichollsi was consistently found in sand dunes. While this sand had favourable water potential properties for buried frogs, the considerable spatial and temporal variation in sand moisture meant that frogs were not always in positive water balance with respect to the surrounding soil. The cocoon-forming N. aquilonius was excavated from two distinct habitat types, a claypan in which frogs had a well-formed cocoon and a dune swale where frogs did not have a cocoon. Cocoons of excavated frogs ranged in thickness from 19.4 microm to 55.61 microm and consisted of 81-229 layers. Cocooned claypan N. aquilonius were nearing exhaustion of their bladder water reserves and had a urine osmolality approaching that of the plasma. By contrast, non-cocooned N. aquilonius from the dune swale were fully hydrated, although soil moisture levels were not as high as calculated to be necessary to maintain water balance. Both species had similar plasma arginine vasotocin (AVT) concentrations ranging from 9.4 to 164 pg ml(-1), except for one cocooned N. aquilonius with a higher concentration of 394 pg ml(-1). For both species, AVT showed no relationship with plasma osmolality over the lower range of plasma osmolalities but was appreciably increased at the highest osmolality recorded. This study provides the first evidence that cocoon formation following burrowing is not obligatory in species that are capable of doing so, but that cocoon formation occurs when soil water conditions are more desiccating than for non-cocooned frogs.

Adaptation, Physiological↗

Pharmacokinetics in the human of a new synthetic vasopressin and oxytocin uterine antagonist.

The pharmacokinetics in the human of 1-deamino-2-D-Tyr(OEt)-4-Thr-8-Orn-vasotocin (dE-TVT), was studied after iv and intranasal administration in 11 subjects at 12 experiments each route. The plasma concentration of the analogue was analysed by means of an arginine vasopressin antibody, which cross-reacted with dE-TVT to 4.7%. When given intravenously as bolus injection (10 nmol/kg/body weight), the total body clearance amounted to 0.623 +/- 0.099 (SEM) l/h kg and the half-life to 16.2 +/- 2.4 min. After intranasal administration (100 nmol/kg/body weight), the bioavailability was 10.5 +/- 2.9%. Peak concentrations in plasma appeared 2-8 min after iv and 10-45 min after intranasal administration. At the end of an observation period of 2 h measurable amounts in plasma were still found in one of the iv and seven of the intranasal experiments. It is concluded that the moderately long half-life is suitable for the treatment of hospitalized patients in premature labour where promising results with intravenous infusion (50 micrograms/min) of dE-TVT have been obtained. It is still uncertain whether or not the absorption of dE-TVT is sufficient for intranasal administration to out-patients with uterine hyperactivity in late pregnancy and to patients with primary dysmenorrhoea, where significant relief of symptoms were seen after iv administration (10 micrograms/kg body weight).

Administration, Intranasal↗

Neurophpophysial hormones and the emission of semen in rabbits.

To elucidate the importance of the neurohypophysial hormones for the emission of semen, several neurohypophysial peptides were tested in male rabbits and the sperm density in the ejaculates was determined. Besides oxytocin and vasopressin, vasotocin and one oxytocin analogue (de-amino1-oxytocin) were used. Only vasopressin, in a dose as low as 10 mU, increased the number of spermatozoa in the ejaculates. It is suggested that vasopressin is of physiological significance for the emission of semen, at least in rabbits.

Animals↗