RESUMO
Parental care represents a suite of distinct behaviors performed by parents to maximize fitness. Dynamic shifts in parental care behaviors, such as between nest defense and direct provisioning of the offspring, are required in response to environmental variation. However, the neural mechanisms which mediate such behavioral shifts remain a mystery. The anemonefish, Amphiprion ocellaris, represents an experimentally valuable model in social neuroscience which is conducive to manipulating the environment while simultaneously measuring parental care. The goal of this study was to determine the extent to which arginine vasotocin (AVT) and isotocin (IT) signaling are necessary for males to shift between direct egg care and aggressive nest defense in the presence of intruders, Domino damselfish (Dascyllus trimaculatus). The IT receptor antagonist desGly-NH2-d(CH2)5[D-Tyr2,Thr4]OVT, significantly reduced direct egg care, while at the same time increased levels of aggressive nest defense relative to vehicle. Conversely, blockade of AVT using the antagonist d(CH2)5[Tyr(Me)2]AVP, reduced aggression and tended to increase egg care. Results demonstrate that male anemonefish alter their parental strategy in response to allospecific intruders, and that IT and AVT signaling oppositely regulate parental care displays of aggression versus egg care.
Assuntos
Agressão/fisiologia , Comportamento de Nidação/fisiologia , Ocitocina/análogos & derivados , Perciformes/fisiologia , Vasotocina/fisiologia , Agressão/efeitos dos fármacos , Animais , Masculino , Comportamento de Nidação/efeitos dos fármacos , Ocitocina/metabolismo , Ocitocina/fisiologia , Receptores de Ocitocina/antagonistas & inibidores , Receptores de Ocitocina/metabolismo , Receptores de Ocitocina/fisiologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia , Comportamento Social , Territorialidade , Vasotocina/análogos & derivados , Vasotocina/antagonistas & inibidores , Vasotocina/metabolismoRESUMO
BACKGROUND: The herbal medicine Bryophyllum pinnatum has been used as a tocolytic agent in anthroposophic medicine and, recently, in conventional settings alone or as an add-on medication with tocolytic agents such as atosiban or nifedipine. We wanted to compare the inhibitory effect of atosiban and nifedipine on human myometrial contractility in vitro in the absence and in the presence of B. pinnatum press juice (BPJ). METHODS: Myometrium biopsies were collected during elective Caesarean sections. Myometrial strips were placed under tension into an organ bath and allowed to contract spontaneously. Test substances alone and at concentrations known to moderately affect contractility in this setup, or in combination, were added to the organ bath, and contractility was recorded throughout the experiments. Changes in the strength (measured as area under the curve (AUC) and amplitude) and frequency of contractions after the addition of all test substances were determined. Cell viability assays were performed with the human myometrium hTERT-C3 and PHM1-41 cell lines. RESULTS: BPJ (2.5 µg/mL), atosiban (0.27 µg/mL), and nifedipine (3 ng/mL), moderately reduced the strength of spontaneous myometrium contractions. When BPJ was added together with atosiban or nifedipine, inhibition of contraction strength was significantly higher than with the tocolytics alone (p = 0.03 and p < 0.001, respectively). In the case of AUC, BPJ plus atosiban promoted a decrease to 48.8 ± 6.3% of initial, whereas BPJ and atosiban alone lowered it to 70.9 ± 4.7% and to 80.9 ± 4.1% of initial, respectively. Also in the case of AUC, BPJ plus nifedipine promoted a decrease to 39.9 ± 4.6% of initial, at the same time that BPJ and nifedipine alone lowered it to 78.9 ± 3.8% and 71.0 ± 3.4% of initial. Amplitude data supported those AUC data. The inhibitory effects of BPJ plus atosiban and of BPJ plus nifedipine on contractions strength were concentration-dependent. None of the test substances, alone or in combination, decreased myometrial cell viability. CONCLUSIONS: BPJ enhances the inhibitory effect of atosiban and nifedipine on the strength of myometrial contractions, without affecting myometrium tissue or cell viability. The combination treatment of BPJ with atosiban or nifedipine has therapeutic potential.
Assuntos
Kalanchoe/química , Miométrio/efeitos dos fármacos , Nifedipino/antagonistas & inibidores , Extratos Vegetais/farmacologia , Nascimento Prematuro/prevenção & controle , Tocolíticos/antagonistas & inibidores , Contração Uterina/efeitos dos fármacos , Vasotocina/análogos & derivados , Adulto , Antagonismo de Drogas , Feminino , Humanos , Técnicas In Vitro , Miométrio/fisiopatologia , Nifedipino/farmacologia , Gravidez , Tocolíticos/farmacologia , Vasotocina/antagonistas & inibidores , Vasotocina/farmacologia , Adulto JovemRESUMO
Vasotocin (VT) is the basic nonapeptide hormone secreted by the neurohypophysis of non-mammalian vertebrates and is involved in the regulation of osmoregulation, metabolism, cardiovascular function, reproduction and behaviour. Among the reproductive function, VT is specifically implicated in final oocyte maturation, ovulation, oviposition/parturition in teleosts, amphibians, reptiles, and birds. The central catecholaminergic system is involved in the regulation of pituitary hormone secretion including gonadotropin, and mediates also changes in environmental photoperiod and temperature. The close apposition of the VT and catecholaminergic systems in the preoptic area of the hypothalamus signifies a strong possibility of their functional interaction. The aim of the present study was to investigate the effects of exogenously administered catecholamines on VT secretion in two different reproductive phases of female catfish Heteropneustes fossilis. For this, the catecholamine precursor L-dihydroxyphenylalanine (L-DOPA) and catecholamines (dopamine-DA, norepinephrine-NE, and epinephrine-E) were intraperitoneally injected in normal catfish and/or along with α-methylparatyrosine (α-MPT, a tyrosine hydroxylase inhibitor). Brain and plasma VT levels were measured by specific ELISA, 24h post injection. Both L-DOPA and DA inhibited brain and plasma VT levels in a concentration-dependent manner in preparatory and prespawning phases. In contrast, NE elicited dose-dependent effects: the lowest dose (0.5 ng/g body mass, BM) was ineffective, the median dose (1 ng/g BM) stimulated, and the high doses (10 and 100 ng/g BM) inhibited VT levels. E stimulated VT levels dose-dependently. A single injection of α-MPT (250 µg/g BM) strongly inhibited VT when given alone and enhanced the inhibitory effects of L-DOPA and DA in the combination groups. The α-MPT inhibition of VT was significantly reduced by the injection of NE (5 ng/g BM) and was restored or elevated by E. When the adrenergic neurotransmitters were given together with α-MPT, the inhibitory effect of the latter was abolished and VT levels were significantly elevated. Thus, the present data indicate that the physiological changes in VT is differentially regulated by the catecholamines (DA inhibits and NE/E stimulates VT).
Assuntos
Catecolaminas/farmacologia , Peixes-Gato/metabolismo , Vasotocina/metabolismo , Animais , Encéfalo/efeitos dos fármacos , Dopamina/farmacologia , Epinefrina/farmacologia , Feminino , Levodopa/farmacologia , Norepinefrina/farmacologia , Reprodução/efeitos dos fármacos , Reprodução/fisiologia , Vasotocina/antagonistas & inibidores , Vasotocina/sangue , Vitelogênese/fisiologia , alfa-Metiltirosina/farmacologiaRESUMO
Previous comparisons of territorial and gregarious finches (family Estrildidae) suggest the hypothesis that arginine vasotocin (VT) neurons in the medial bed nucleus of the stria terminalis (BSTm) and V(1a)-like receptors in the lateral septum (LS) promote flocking behavior. Consistent with this hypothesis, we now show that intraseptal infusions of a V(1a) antagonist in male zebra finches (Taeniopygia guttata) reduce gregariousness (preference for a group of 10 versus 2 conspecific males), but have no effect on the amount of time that subjects spend in close proximity to other birds ("contact time"). The antagonist also produces a profound increase in anxiety-like behavior, as exhibited by an increased latency to feed in a novelty-suppressed feeding test. Bilateral knockdown of VT production in the BSTm using LNA-modified antisense oligonucleotides likewise produces increases in anxiety-like behavior and a potent reduction in gregariousness, relative to subjects receiving scrambled oligonucleotides. The antisense oligonucleotides also produced a modest increase in contact time, irrespective of group size. Together, these combined experiments provide clear evidence that endogenous VT promotes preferences for larger flock sizes, and does so in a manner that is coupled to general anxiolysis. Given that homologous peptide circuitry of the BSTm-LS is found across all tetrapod vertebrate classes, these findings may be predictive for other highly gregarious species.
Assuntos
Comportamento Animal/fisiologia , Comportamento Exploratório/fisiologia , Receptores de Vasopressinas/fisiologia , Septo do Cérebro/fisiologia , Aves Canoras/fisiologia , Vasotocina/fisiologia , Animais , Ansiedade/induzido quimicamente , Comportamento Animal/efeitos dos fármacos , Comportamento Exploratório/efeitos dos fármacos , Masculino , Oligonucleotídeos Antissenso/administração & dosagem , Septo do Cérebro/efeitos dos fármacos , Vasotocina/antagonistas & inibidoresRESUMO
Cultured cells from pineal glands of human fetuses release into their media a substance that has antidiuretic and hydroosmotic activities. The ratio of these activities as well as their susceptibility to tryptic digestion, specific oxidative inactivation by tyrosinase, and reductive inactivation by sodium thioglycollate, indicates the presence of a basic peptide, presumably identical with arginine vasotocin. The total amount of this peptide released into the culture media during 38 days of incubation is about ten times greater than the amount contained in nonincubated pineal glands from fetuses of the same age, strongly suggesting that fetal ependymal cells from the pineal gland can synthesize in vitro a peptide similar to arginine vasotocin.
Assuntos
Glândula Pineal/metabolismo , Vasotocina/biossíntese , Animais , Anuros , Arginina , Bioensaio , Catecol Oxidase/farmacologia , Células Cultivadas , Diurese/efeitos dos fármacos , Feminino , Feto , Idade Gestacional , Humanos , Masculino , Osmose/efeitos dos fármacos , Glândula Pineal/embriologia , Rana temporaria , Ratos , Tripsina/farmacologia , Bexiga Urinária/efeitos dos fármacos , Útero/efeitos dos fármacos , Vasotocina/antagonistas & inibidores , Vasotocina/farmacologiaRESUMO
Arginine vasotocin (VT), and its mammalian homologue arginine vasopressin (VP), are neuropeptides involved in the regulation of social behaviors and stress responsiveness. Previous research has demonstrated opposing effects of VT/VP on aggression in different species. However, these divergent effects were obtained in different social contexts, leading to the hypothesis that different populations of VT/VP neurons regulate behaviors in a context-dependent manner. We here use VP antagonists to block endogenous VT function in male zebra finches (Taeniopygia guttata) within a semi-natural, mixed-sex colony setting. We examine the role of VT in the regulation of aggression and courtship, and in pair bond formation and maintenance, over the course of three days. Although our results confirm previous findings, in that antagonist treatment reduces aggressive mate competition during an initial behavioral session during which males encounter novel females, we find that the treatment effects are completely reversed within hours of colony establishment, and the antagonist treatment instead facilitates aggression in later sessions. This reversal occurs as aggression shifts from mate competition to nest defense, but is not causally associated with pairing status per se. Instead, we hypothesize that these divergent effects reflect context-specific activation of hypothalamic and amygdalar VT neurons that exert opposing influences on aggression. Across contexts, effects were highly specific to aggression and the antagonist treatment clearly failed to alter latency to pair bond formation, pair bond stability, and courtship. However, VT may still potentially influence these behaviors via promiscuous oxytocin-like receptors, which are widely distributed in the zebra finch brain.
Assuntos
Meio Ambiente , Abrigo para Animais , Comportamento Sexual Animal/fisiologia , Comportamento Social , Vasotocina/metabolismo , Agressão/fisiologia , Análise de Variância , Animais , Cateterismo , Corte , Tentilhões , Masculino , Ligação do Par , Fatores de Tempo , Vasotocina/antagonistas & inibidoresRESUMO
Cooperation between unrelated individuals usually involves investments that often mean a decrease in immediate payoffs, but ensure future benefits. Here we investigated the potential role of the neuropeptides Arginine-vasotocin (AVT) and Isotocin (IT) as proximate agents affecting individuals' cooperative levels in the Indo-pacific bluestreak cleaner wrasse Labroides dimidiatus. Their 'client' reef fish partners only benefit from interacting if cleaners eat ectoparasites and refrain from gleaning preferred client mucus. Thus, cleaners must control their impulse to eat according to their preference, and eat less preferred items to maintain ongoing interactions and avoid clients' leaving or punishing. We found that solely the experimental transient higher dosage of AVT led to a decrease of cleaners' willingness to feed against their preference, while IT and AVT antagonists had no significant effects. The sole effect of AVT on cleaner's performance may imply a link between AVT's influence and a potential activation of a stress response. Our results confirm the importance of the AVT/AVP system as an agent affecting levels of cooperation, offering a potential mechanistic pathway for the reported flexible service quality that cleaners provide their clients.
Assuntos
Comportamento Cooperativo , Comportamento Alimentar/fisiologia , Ocitocina/análogos & derivados , Perciformes/fisiologia , Vasotocina/metabolismo , Animais , Tomada de Decisões/efeitos dos fármacos , Tomada de Decisões/fisiologia , Comportamento Alimentar/efeitos dos fármacos , Preferências Alimentares/efeitos dos fármacos , Preferências Alimentares/fisiologia , Aprendizagem/efeitos dos fármacos , Aprendizagem/fisiologia , Ocitocina/antagonistas & inibidores , Ocitocina/metabolismo , Simbiose/fisiologia , Vasotocina/antagonistas & inibidoresRESUMO
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.
Assuntos
Toxinas Bacterianas/farmacologia , Testículo/fisiologia , Vasotocina/farmacologia , Toxina Adenilato Ciclase , Animais , Células Cultivadas , Gonadotropina Coriônica/farmacologia , AMP Cíclico/metabolismo , Masculino , Toxina Pertussis , Ratos , Testículo/citologia , Testículo/metabolismo , Testosterona/metabolismo , Vasotocina/antagonistas & inibidores , Fatores de Virulência de BordetellaRESUMO
The authors measured the effects of centrally infused peptides on social approach behaviors in goldfish (Carassius auratus), a social teleost. Vasotocin (VT) inhibited approach responses toward the visual stimuli of conspecifics in the absence of aggressive or sexual olfactory contextual cues in males, and a V1 receptor antagonist stimulated such responses, at least in males that were not highly social in baseline conditions, as did isotocin (IT). In the absence of social stimuli, VT did not affect activity, therefore indicating that the inhibition was not the result of nonspecific effects on arousal or motor functioning. These experiments indicate that VT and IT induce opposite effects on social approach responses in male goldfish and that endogenous VT, at least, is associated with levels of sociality.
Assuntos
Arginina Vasopressina/análogos & derivados , Comportamento Animal/efeitos dos fármacos , Ocitocina/análogos & derivados , Ocitocina/farmacologia , Comportamento Social , Vasotocina/farmacologia , Animais , Arginina Vasopressina/farmacologia , Relação Dose-Resposta a Droga , Carpa Dourada , Locomoção/efeitos dos fármacos , Masculino , Estimulação Luminosa/métodos , Atrativos Sexuais/farmacologia , Comportamento Sexual Animal , Fatores de Tempo , Vasotocina/antagonistas & inibidoresRESUMO
The effect of rat atrial natriuretic peptide (rANP) on hormonal stimulated osmotic water permeability (Jw, hydrosmotic effect) and net ion transport (short-circuit current, SCC, natriferic effect) was studied on toad skin, a tissue with functional similarities to the mammalian distal nephron, in order to assess actions on transport mechanisms. Rat atrial natriuretic peptide, rANP-99-126 (rANP) inhibited stimulated SCC and Jw to submaximal concentrations of arginine vasotocin (AVT) at a site before cyclic AMP generation. The angiotensin-converting enzyme inhibitor (ACEI) MK-422 did not modify the inhibitory effect of ANP in the stimulated Jw.
Assuntos
Fator Natriurético Atrial/farmacologia , Pele/efeitos dos fármacos , Vasotocina/antagonistas & inibidores , Animais , Bucladesina/farmacologia , Bufo arenarum , AMP Cíclico/biossíntese , Enalaprilato/farmacologia , Feminino , Transporte de Íons/efeitos dos fármacos , Masculino , Osmose/efeitos dos fármacos , Permeabilidade/efeitos dos fármacos , Ratos , Pele/metabolismo , Sódio/metabolismo , Teofilina/farmacologia , Vasotocina/farmacologiaRESUMO
The purpose of the present study was to assess the effects of cyclosporine A (CyA) in the osmotic water flow response of isolated toad bladder to arginine-vasotocin (AVT) and to angiotensin II (Ang II) and AVT in isolated toad skin. CyA added to the dermal side of isolated toad skin or to the serosal side of toad bladder in concentrations of 0.42. 10(-6) M to 0.42. 10(-7) M had no effect on basal osmotic water permeability (Posm) but inhibited the hormonal response to AVT in both membranes (AVT 10(-10) M in toad bladder and 10(-8) to 10(-9) M in toad skin). CyA also inhibited the Posm response to Ang II (10(-7) M) in toad skin in concentrations of 0.42. 10(-6) M and 0.42. 10(-7) M. In toad bladder it could be demonstrated that the inhibitory effect was reversible. CyA in concentrations of 0.42. 10(-6) M inhibited the Posm response of toad skin to theophylline (3.2. 10(-3) M) and to dibutyryl cyclic AMP (6.3. 10(-3) M) suggesting an effect distal to the generation of cyclic AMP. These responses would support the possibility of a diuretic effect in the mammalian nephron.
Assuntos
Angiotensina II/antagonistas & inibidores , Ciclosporina/farmacologia , Pele/efeitos dos fármacos , Pele/metabolismo , Bexiga Urinária/efeitos dos fármacos , Bexiga Urinária/metabolismo , Vasotocina/antagonistas & inibidores , Água/metabolismo , Animais , Bufo arenarum , Técnicas In Vitro , Osmose/efeitos dos fármacos , Permeabilidade/efeitos dos fármacosRESUMO
Many marine fishes change sex in response to social cues when the dominance hierarchy is perturbed. Arginine-vasotocin (AVT) and the mammalian homolog arginine vasopressin are neuropeptides involved in social and reproductive behaviors across vertebrate taxa. The goal of this study was to determine whether AVT signaling influences aggression and expression of c-Fos, a marker of neuroplasticity, in key brain regions of the social decision circuit in Amphiprion ocellaris clownfish, a species where behavioral dominance precedes gonadal sex change from male to female. In experiment 1, juvenile clownfish (average mass 2.5g) were paired together in a tank (a total of 24 pairs), matched approximately for size with one fish randomly receiving either an intraperitoneal injection of the arginine vasopressin V1a receptor antagonist (Manning compound) or saline vehicle, and evaluated for aggressive and submissive behaviors over a 10-min period. The second experiment was a repeat of the first using five pairs of mature, reproductive males, except the animals interacted for 90-min immediately followed by euthanasia for immunohistochemical detection of c-Fos protein. Numbers of c-Fos-positive cells were quantified in the preoptic area of the hypothalamus (POA), the anterior tuberal nucleus (aTn), and periventricular nucleus of the posterior tuberculum (TPp). Manning compound significantly reduced aggression and the probability of winning the contest relative to saline (vehicle) controls. In experiment 2, saline-treated fish displayed approximately twice as many c-Fos-positive cells in the POA and 25% more in the TPp than the Manning-treated fish, no differences were observed in the aTn. Taken together, results suggest AVT signaling is necessary for aggressive behavior and expression of neuroplasticity in the POA and TPp that likely contributes to behavioral dominance and hence, sex change in A. ocellaris.
Assuntos
Hipotálamo Posterior/metabolismo , Área Pré-Óptica/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismo , Transdução de Sinais/fisiologia , Vasotocina/metabolismo , Fatores Etários , Agressão/fisiologia , Animais , Arginina Vasopressina/análogos & derivados , Arginina Vasopressina/farmacologia , Feminino , Peixes , Hierarquia Social , Masculino , Transdução de Sinais/efeitos dos fármacos , Fatores de Tempo , Vasotocina/antagonistas & inibidoresAssuntos
Arginina Vasopressina/fisiologia , Receptores de Vasopressinas , Testículo/fisiologia , Androgênios/biossíntese , Animais , Arginina Vasopressina/antagonistas & inibidores , Arginina Vasopressina/genética , Arginina Vasopressina/farmacologia , Bucladesina/farmacologia , Células Cultivadas , Gonadotropina Coriônica/metabolismo , Gonadotropina Coriônica/farmacologia , AMP Cíclico/metabolismo , Cistinil Aminopeptidase/metabolismo , Feminino , Hormônio Liberador de Gonadotropina/farmacologia , Hipotálamo/metabolismo , Células Intersticiais do Testículo/fisiologia , Hormônio Luteinizante/fisiologia , Lipressina/farmacologia , Masculino , Ovário/fisiologia , Hormônios Neuro-Hipofisários/fisiologia , Precursores de Proteínas/metabolismo , Processamento de Proteína Pós-Traducional , RNA Mensageiro/metabolismo , Ratos , Ratos Brattleboro , Receptores de Angiotensina/fisiologia , Receptores de Superfície Celular/fisiologia , Receptores LHRH , Células de Sertoli/fisiologia , Testículo/efeitos dos fármacos , Vasotocina/antagonistas & inibidores , Vasotocina/farmacologiaRESUMO
RATIONALE: Oxytocin (OT) and arginine-vasopressin (AVP) regulate social behavior in mammals. Zebrafish (Danio rerio) allows higher throughput and ease in studying human brain disorders. OBJECTIVES: This study investigated in zebrafish the effect of non-mammalian homologs isotocin (IT) and vasotocin (AVT) in comparison with OT/AVP on social behavior and fear response to predator. The mechanism was studied using the most human selective OT and AVP receptor antagonists. METHODS: Zebrafish were injected i.m. with increasing doses (0.001-40 ng/kg) of the neuropeptides. DesGly-NH(2)-d(CH(2))(5)-[D-Tyr(2),Thr(4)]OVT) for OT receptor, SR 49059 for V1a subtype receptor, and SSR-149415 for V1b subtype receptor were injected i.m. 10 min before each agonist. RESULTS: All the peptides increased social preference and reduced fear to predator response in a dose-dependent manner interpolated by symmetrical parabolas. AVT/AVP were more potent to elicit anxiolytic than social effect while IT and OT were equally potent. All the antagonists dose-dependently inhibited both the effects induced by the neuropeptides. The ratio between the ED50 obtained for blocking the OT-induced effects on social preference and fear response to predator was very high only for desglyDTTyrOVT (160). SR49059 showed the highest ratio in blocking AVP-induced effects (807). The less selective antagonist appeared to be SSR149415. CONCLUSIONS: For the first time, IT/AVT and OT/AVP were found to modulate in zebrafish, social behavior, unrelated to sex, and fear to predator response through at least two different receptors. Zebrafish is confirmed as a valid, reliable model to study deficit in social behavior characteristic of some psychiatric disorders.
Assuntos
Ansiolíticos/farmacologia , Medo/efeitos dos fármacos , Ocitocina/fisiologia , Receptores de Vasopressinas/fisiologia , Comportamento Social , Vasopressinas/fisiologia , Vasotocina/fisiologia , Animais , Antagonistas dos Receptores de Hormônios Antidiuréticos , Relação Dose-Resposta a Droga , Indóis/farmacologia , Ornipressina/análogos & derivados , Ornipressina/farmacologia , Ocitocina/análogos & derivados , Ocitocina/antagonistas & inibidores , Ocitocina/farmacologia , Hormônios Neuro-Hipofisários/farmacologia , Hormônios Neuro-Hipofisários/fisiologia , Pirrolidinas/farmacologia , Ensaio Radioligante/métodos , Ensaio Radioligante/estatística & dados numéricos , Receptores de Ocitocina/antagonistas & inibidores , Receptores de Ocitocina/fisiologia , Natação , Vasopressinas/antagonistas & inibidores , Vasopressinas/farmacologia , Vasotocina/antagonistas & inibidores , Vasotocina/farmacologia , Peixe-ZebraAssuntos
Probenecid/farmacologia , Bexiga Urinária/efeitos dos fármacos , Água/metabolismo , Animais , Anuros , Arginina/farmacologia , Transporte Biológico/efeitos dos fármacos , AMP Cíclico/farmacologia , Antagonismo de Drogas , Condutividade Elétrica/efeitos dos fármacos , Ocitocina/antagonistas & inibidores , Ocitocina/farmacologia , Permeabilidade , Rana esculenta , Estimulação Química , Teofilina/farmacologia , Bexiga Urinária/metabolismo , Vasotocina/antagonistas & inibidores , Vasotocina/farmacologiaRESUMO
We have previously demonstrated that centrally administered vasotocin (VT) inhibits social approach toward same-sex conspecifics in male and female goldfish, and that this behavioral effect is dependent upon VT projections to the hindbrain. We now show that there are no sex differences in sensitivity to the behavioral effects of VT, though differences do exist in responsiveness across seasons in both sexes. A central dose of 1 microg, but not 200 ng, inhibited social approach in goldfish in non-reproductive condition, whereas a dose as low as 40 ng inhibited social approach in fish in full reproductive condition. In males and females in full reproductive condition, social approach behavior was facilitated by central administration of 500 ng of a V(1A) specific antagonist. In addition, the behavioral effects of exogenously administered central VT were blocked by central administration of 1 microg of a V(1A) antagonist. These results demonstrate that the propensity to approach a conspecific, a simple behavior underlying many social interactions, is controlled by a V(1A)-like receptor, and that VT's behavioral effects depend on reproductive context. Quantitative real-time PCR showed that the seasonal changes in behavioral responsiveness to VT are associated with changes in the expression of a V(1A)-like receptor in the hindbrain, but not the mid- or forebrain, indicating that the seasonal regulation of social approach behavior likely depends on the local modulation of the expression of this receptor within a primitive peptide circuit in this species.
Assuntos
Comportamento Animal/efeitos dos fármacos , Rombencéfalo/efeitos dos fármacos , Estações do Ano , Vasotocina/metabolismo , Vasotocina/farmacologia , Análise de Variância , Animais , Comportamento Animal/fisiologia , Relação Dose-Resposta a Droga , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/fisiologia , Carpa Dourada/fisiologia , Injeções Intraventriculares/métodos , Masculino , Receptores de Vasopressinas/genética , Receptores de Vasopressinas/metabolismo , Rombencéfalo/fisiologia , Fatores Sexuais , Comportamento Social , Vasotocina/agonistas , Vasotocina/antagonistas & inibidoresRESUMO
Since Gram-negative bacteria are known to be present in the cavity of urinary bladders in amphibian species, it was interesting to study the effect of bacterial endotoxins on epithelial signaling network which provides the arginine-vasotocin-induced increase of osmotic water permeability (OWP). The effect of LPS E. coli on AVT-induced OWP was studied in isolated frog Rana temporaria L. urinary bladder incubated during 20-21 hours in modified L-15 culture medium in sterile conditions. The LPS (25 microg/ml) was added into the mucosal solution. It was shown that exposure to LPS caused a strong suppression of the increase of OWP under AVT (0.5 nM), forskolin (35 microM) or IBMX (200 microM). Moreover, LPS induced more than 2-folds decrease both ofbasal and AVT-stimulated content of cAMP in the bladder tissue. The inhibitory effect of LPS on AVT-induced increase of OWP was eliminated in the presence of ODQ, 20 microM, a cytosolic guanylate cyclase inhibitor. With the use of RT-PCR it was shown that the expression of mRNA iNOS was 10-fold increased in 6 hours after LPS administration. These findings demonstrate the ability of frog bladder mucosal epithelial cells to recognize bacterial LPS and initiate antipathogen immune response related to increased production of nitric oxide. The activation of signal transduction cascade mediated by the LPS-induced immune response leads to a decrease of intracellular cAMP and down-regulates AVT-stimulated OWP acting at least in part through NO/cGMP-dependent signaling pathway.
Assuntos
Escherichia coli/imunologia , Lipopolissacarídeos/imunologia , Osmose , Bexiga Urinária/microbiologia , Bexiga Urinária/fisiologia , Animais , AMP Cíclico/análise , GMP Cíclico/análise , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Lipopolissacarídeos/farmacologia , Masculino , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Osmose/efeitos dos fármacos , Rana temporaria , Bexiga Urinária/efeitos dos fármacos , Vasotocina/antagonistas & inibidores , Vasotocina/farmacologia , Água/metabolismoRESUMO
Renal facilitative urea transporters play a vital role in the urinary concentrating mechanism. UT-A3 is a phloretin-sensitive urea transporter that in the mouse is expressed on the basolateral membrane of renal inner medullary collecting duct (IMCD) cells. In this study, we engineered a Madin-Darby canine kidney (MDCK) I cell line that stably expresses mouse UT-A3 (MDCK-mUT-A3). Immunoblotting using the UT-A-targeted antibody ML446 detected a approximately 40-kDa signal in MDCK-mUT-A3 protein that corresponds to mUT-A3. Using cultured epithelial monolayers, radioactive (14)C-urea flux experiments determined that basolateral urea transport was no different between MDCK-mUT-A3 and control MDCK-FLZ cells under basal conditions [not significant (NS), ANOVA]. However, exposure to arginine vasopressin (AVP) significantly stimulated basolateral urea flux in MDCK-mUT-A3 monolayers (P < 0.05, ANOVA), while it had no effect in control MDCK-FLZ monolayers (NS, ANOVA). The AVP-stimulated basolateral urea transport in MDCK-mUT-A3 was inhibited by 1,3 dimethyl urea (P < 0.05, ANOVA) or phloretin (P < 0.05, ANOVA), both known inhibitors of facilitative urea transporters. MDCK-mUT-A3 basolateral urea flux was also stimulated by increasing intracellular levels of cAMP, via forskolin (P < 0.05, ANOVA), or intracellular calcium, via ATP (P < 0.05, ANOVA). Finally, 1-h preincubation with a specific PKA inhibitor, H89, significantly inhibited the increase in urea transport produced by AVP (P < 0.05, ANOVA). In conclusion, we have produced the first renal cell line to stably express the mUT-A3 urea transporter. Our results indicate that mUT-A3 is acutely regulated by AVP, via a PKA-dependent pathway. These findings have important implications for the regulation of urea transport in the renal IMCD and the urinary concentrating mechanism.
Assuntos
Proteínas de Membrana Transportadoras/biossíntese , Animais , Cálcio/farmacologia , Linhagem Celular , AMP Cíclico/farmacologia , Cães , Regulação da Expressão Gênica/efeitos dos fármacos , Immunoblotting , Isoquinolinas/farmacologia , Proteínas de Membrana Transportadoras/efeitos dos fármacos , Proteínas de Membrana Transportadoras/fisiologia , Compostos de Metilureia/farmacologia , Camundongos , Oócitos/efeitos dos fármacos , Floretina/farmacologia , Inibidores de Proteínas Quinases/farmacologia , Sulfonamidas/farmacologia , Transfecção , Ureia/metabolismo , Vasotocina/antagonistas & inibidores , Vasotocina/farmacologia , Xenopus laevis , Transportadores de UreiaRESUMO
PGE(2) is a well-known inhibitor of the antidiuretic hormone-induced increase of osmotic water permeability (OWP) in different osmoregulatory epithelia; however, the mechanisms underlying this effect of PGE(2) are not completely understood. Here, we report that, in the frog Rana temporaria urinary bladder, EP(1)-receptor-mediated inhibition of arginine-vasotocin (AVT)-induced OWP by PGE(2) is attributed to increased generation of nitric oxide (NO) in epithelial cells. It was shown that the inhibitory effect of 17-phenyl-trinor-PGE(2) (17-ph-PGE(2)), an EP(1) agonist, on AVT-induced OWP was significantly reduced in the presence of 7-nitroindazole (7-NI), a neuronal NO synthase (nNOS) inhibitor. NO synthase (NOS) activity in both lysed and intact epithelial cells measured as a rate of conversion of l-[(3)H]arginine to l-[(3)H]citrulline was Ca(2+) dependent and inhibited by 7-NI. PGE(2) and 17-ph-PGE(2), but not M&B-28767 (EP(3) agonist) or butaprost (EP(2) agonist), stimulated NOS activity in epithelial cells. The above effect of PGE(2) was abolished in the presence of SC-19220, an EP(1) antagonist. 7-NI reduced the stimulatory effect of 17-ph-PGE(2) on NOS activity. 17-ph-PGE(2) increased intracellular Ca(2+) concentration and cGMP in epithelial cells. Western blot analysis revealed an nNOS expression in epithelial cells. These results show that the inhibitory effect of PGE(2) on AVT-induced OWP in the frog urinary bladder is based at least partly on EP(1)-receptor-mediated activation of the NO/cGMP pathway, suggesting a novel cross talk between AVT, PGE(2), and nNOS that may be important in the regulation of water transport.
Assuntos
AMP Cíclico/fisiologia , Dinoprostona/farmacologia , Óxido Nítrico/fisiologia , Ocitócicos/farmacologia , Receptores de Prostaglandina E/fisiologia , Transdução de Sinais/efeitos dos fármacos , Bexiga Urinária/efeitos dos fármacos , Bexiga Urinária/metabolismo , Vasotocina/antagonistas & inibidores , Vasotocina/farmacologia , Equilíbrio Hidroeletrolítico/efeitos dos fármacos , Animais , Western Blotting , Química Encefálica/efeitos dos fármacos , Cálcio/metabolismo , Ácido Dibenzo(b,f)(1,4)oxazepina-10(11H)-carboxílico, 8-cloro-, 2-acetilidrazida/farmacologia , Dinoprostona/análogos & derivados , Ativação Enzimática/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Técnicas In Vitro , Indazóis/farmacologia , Masculino , Óxido Nítrico Sintase Tipo I/antagonistas & inibidores , Óxido Nítrico Sintase Tipo I/metabolismo , Permeabilidade/efeitos dos fármacos , Rana temporaria , Receptores de Prostaglandina E Subtipo EP1RESUMO
Uteri of Anolis carolinensis exhibited spontaneous rhythmic contractions in vitro. Addition of arginine vasotocin (AVT) caused an immediate, strong, tonic contraction followed by rhythmic contractions with the same frequency as spontaneous contractions but of a greater amplitude. At low tension (1.5 g) the AVT-induced tonic contraction was blocked by low dose of indomethacin, suggesting that it is influenced by calcium rather than prostaglandins (PGs). An increase in tension (from 1.5 to 15 g) reduced the duration of the AVT-induced tonic contraction; this stretch-induced decrease was also blocked by indomethacin. Stretch also decreased the duration of the rhythmic contractions, but this stretch effect was not inhibited by indomethacin. The rest interval between rhythmic contractions was decreased by PGF2alpha and PGE2, and indomethacin or stretch blocked these PG effects. Indomethacin, AVT, or stretch alone did not affect PGF2alpha secretion from AVT-treated uteri. Stretch also reduced PGF2alpha secretion from AVT-treated uteri, an effect inhibited by indomethacin.