RESUMO
Zebrafish exhibit fear contagion, a basic form of empathy, and when observing social fellows that have been exposed to predation cues, will themselves exhibit similar distress behaviours. As in mammals, the nonapeptide hormone oxytocin is essential for this empathic response, and homologous areas of the brain are involved, suggesting that the mechanistic basis of empathy may be conserved among vertebrates.
RESUMO
Social buffering of stress refers to the effect of a social partner in reducing the cortisol or corticosterone response to a stressor. It has been well studied in mammals, particularly those that form pair bonds. Recent studies on fishes suggest that social buffering of stress also occurs in solitary species, gregarious species that form loose aggregations and species with well-defined social structures and bonds. The diversity of social contexts in which stress buffering has been observed in fishes holds promise to shed light on the evolution of this phenomenon among vertebrates. Equally, the relative simplicity of the fish brain is advantageous for identifying the neural mechanisms responsible for social buffering. In particular, fishes have a relatively small and simple forebrain but the brain regions that are key to social buffering, including the social behaviour network, the amygdala and the hypothalamic-pituitary-adrenal/interrenal axis, are functionally conserved across vertebrates. Thus, we suggest that insight into the mechanistic and evolutionary underpinnings of stress buffering in vertebrates can be gained from the study of social buffering of stress in fishes.
Assuntos
Corticosterona , Hidrocortisona , Animais , Corticosterona/farmacologia , Sistema Hipófise-Suprarrenal/fisiologia , Sistema Hipotálamo-Hipofisário/fisiologia , Peixes/fisiologia , Estresse Psicológico , MamíferosRESUMO
European sea bass (Dicentrarchus labrax) are a major aquaculture species that live in habitats with fluctuating salinities that are sometimes higher than in seawater (SW). Atlantic and West-Mediterranean genetic lineages were compared regarding intestinal neuropeptide receptor expression in SW (36%) and following a two-week transfer to hypersalinity (HW, 55%). Phylogenetic analysis revealed seven neuropeptide receptors belonging to the arginine vasotocine (AVTR) family and two isotocin receptors (ITR). Among AVTR paralogs, the highest mRNA levels were recorded for v1a2, with a two- to fourfold upregulation in the European sea bass intestinal sections after transfer of fish to HW. Principal component analysis in posterior intestines showed that v1a2 expression grouped together with the expression and activity of main ion transporters and channels involved in solute-coupled water uptake, indicating a possible role of this receptor in triggering water absorption. v1a1 expression, however, was decreased or did not change after transfer to hypersaline water. Among ITR paralogs, itr1 was the most expressed paralog in the intestine and opposite expression patterns were observed following salinity transfer, comparing intestinal sections. Overall, different expression profiles were observed between genetic lineages for several analyzed genes which could contribute to different osmotic stress-related responses in D. labrax lineages.
Assuntos
Bass , Animais , Bass/genética , Bass/metabolismo , Filogenia , Brânquias/metabolismo , Intestinos , Água do Mar , Água/metabolismoRESUMO
The synthesis and release of LH and FSH in the pituitary of vertebrates are differentially regulated during gonadal development and maturation. However, the underlying neuroendocrine mechanisms remain to be fully elucidated. The present study examined the possible involvement of isotocin (Ist), an oxytocin-like neuropeptide, in the regulation of Lh and Fsh in a teleost, the ricefield eel Monopterus albus. The immunoreactive isotocin receptor 2 (Istr2) was shown to be localized to Lh but not Fsh cells. In contrast, immunoreactive isotocin receptor 1 (Istr1) was not observed in either Lh or Fsh cells in the pituitary. Interestingly, Lh cells in female ricefield eels expressed Istr2 and secreted Lh in response to Ist challenge stage-dependently and in correlation with ovarian vitellogenesis. Moreover, Ist decreased Lh contents in the pituitary of female fish, indicating its stimulatory roles on Lh release in vivo. The induction of Lh release by Ist in dispersed pituitary cells was blocked by a PLC or IP3R inhibitor but not by a PKA or PKC inhibitor, indicating the involvement of the IP3/Ca2+ pathway. Collectively, the above results indicate that isotocin may bind to Istr2 to stimulate Lh release via the IP3/Ca2+ pathway, and play important roles in the ovarian maturation in ricefield eels. Furthermore, the present study suggests a novel neuroendocrine mechanism underlying the differential regulation of Lh and Fsh in vertebrates.
Assuntos
Enguias/fisiologia , Hormônio Foliculoestimulante/metabolismo , Hormônio Luteinizante/metabolismo , Ocitocina/metabolismo , Animais , Feminino , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Masculino , Transporte Proteico , Maturidade SexualRESUMO
The vasopressin-vasotocin (AVP-AVT) and oxytocin-mesotocin-isotocin (OT-MT-IT) families of nonapeptides are of great importance in shaping context-dependent modulations of a conserved and yet highly plastic network of brain areas involved in social behavior: the social behavior network. The nonapeptide systems of teleost fish are highly conserved and share a common general organization. In this study, we first describe the presence of IT cells and projections in the brain of an electric fish, Gymnotus omarorum. Second, we confirm that IT neuron types and distribution in the preoptic area (POA) follow the same general pattern previously described in other teleost species. Third, we show that although IT and AVT neurons occur intermingled within the POA of G. omarorum and can be classified into the same subgroups, they present subtle but remarkable differences in size, number, and location. Finally, we show that unlike AVT, IT has no effect on basal electric signaling, reinforcing the specificity in the actions that each one of these nonapeptides has on social behavior and communication.
Assuntos
Peixe Elétrico , Vasotocina , Animais , Peixe Elétrico/fisiologia , Neurônios , Ocitocina/análogos & derivadosRESUMO
Isotocin controls ion regulation through modulating the functions of ionocytes (also called mitochondria-rich cells or chloride cells). However, little is known about the upstream molecule of the isotocin system. Herein, we identify transient receptor potential vanilloid 4 (TRPV4), which regulates the mRNA and protein expressions of isotocin and affects ion regulation through the isotocin pathway. Double immunohistochemical results showed that TRPV4 is expressed in isotocinergic neurons in the hypothalamus of the adult zebrafish brain. To further elucidate the roles of TRPV4, we manipulated TRPV4 protein expression and evaluated its ionoregulatory functions in zebrafish embryos. TRPV4 gene knockdown with morpholino oligonucleotides decreased ionic contents (Na+, Cl-, and Ca2+) of whole larvae and the H+-secreting function of larval skin of zebrafish. mRNA expressions of ionocyte-related transporters, including H+-ATPase, the epithelial Ca2+ channel, and the Na+-Cl- cotransporter, were also suppressed in trpv4 morphants. Numbers of ionocytes (H+-ATPase-rich cells and Na+-K+-ATPase-rich cells) and epidermal stem cells in zebrafish larval skin also decreased after trpv4 knockdown. Our results showed that TRPV4 modulates ion balance through the isotocin pathway.
Assuntos
Ocitocina/análogos & derivados , Canais de Cátion TRPV/metabolismo , Proteínas de Peixe-Zebra/metabolismo , Animais , Animais Geneticamente Modificados , Cálcio/metabolismo , Cloretos/metabolismo , Regulação da Expressão Gênica/fisiologia , Transporte de Íons , Larva , Neurônios , Ocitocina/genética , Ocitocina/metabolismo , Sódio/metabolismo , Canais de Cátion TRPV/genética , Equilíbrio Hidroeletrolítico , Peixe-Zebra , Proteínas de Peixe-Zebra/genéticaRESUMO
Pro-social effects of oxytocin and its homologues are well-documented in birds and mammals. However, in fishes, the effect of isotocin, the homologue of oxytocin, on social behaviour is less clear. Studies in fishes have generally shown no effect of isotocin on social behaviours or even an anti-social effect. In our study, we measured association preference for conspecifics in 92 adult guppies (46 females and 46 males), half of which were injected with isotocin and the other half with an isotocin antagonist. We found that individuals injected with isotocin spent 29% more time associating with conspecifics than individuals injected with an isotocin antagonist. The effect of isotocin on association time did not differ between males and females. Our study provides some of the first evidence of a pro-social effects of isotocin in a fish and suggests that in fishes, isotocin may have a homologous role to oxytocin, at least in promoting shoaling behaviour.
Assuntos
Poecilia , Animais , Feminino , Masculino , Ocitocina/análogos & derivados , Comportamento SocialRESUMO
On the wild spawning grounds, the round gobies Neogobius melanostomus are subjected to different social cues, such as sex-separation and high fish density. We designed an experiment to stimulate natural social stress when fish are separated from opposite sex individuals and exposed to close proximity of same-sex conspecifics. We examined the effects of different sex compositions on aggressiveness and brain concentrations of arginine vasotocin (AVT) and isotocin (IT), as AVT and IT are known to be involved in aggressive interactions during reproduction. The round gobies were kept in three experimental groups: same-sex groups broken down into male-only and female-only groups and mixed-sex groups. In this study, males and females from same-sex groups showed overt aggression and competition. Separation stress stimulated aggressive responses in both sexes, but the link between brain AVT and IT concentration and aggressive behavior was evident only in male-only group. In the male-only group, AVT and IT levels were the highest. This study shows that sex composition of the social environment can affect aggressive behavior as well as AVT and IT concentration in the whole brain of the round goby.
Assuntos
Comportamento Animal/fisiologia , Encéfalo/metabolismo , Ocitocina/análogos & derivados , Perciformes/fisiologia , Vasotocina/metabolismo , Agressão , Animais , Feminino , Masculino , Ocitocina/metabolismo , Reprodução , Meio Social , Estresse PsicológicoRESUMO
Living in groups affords individuals many benefits, including the opportunity to reduce stress. In mammals, such 'social buffering' of stress is mediated by affiliative relationships and production of the neuropeptide oxytocin, but whether these mechanisms facilitate social buffering across vertebrates remains an open question. Therefore, we evaluated whether the social environment influenced the behavioural and physiological recovery from an acute stressor in a group-living cichlid, Neolamprologus pulcher. Individual fish that recovered with their social group displayed lower cortisol levels than individuals that recovered alone. This social buffering of the stress response was associated with a tendency towards lower transcript abundance of arginine vasotocin and isotocin in the preoptic area of the brain, suggesting reduced neural activation of the stress axis. Individuals that recovered with their social group quickly resumed normal behaviour but received fewer affiliative acts following the stressor. Further experiments revealed similar cortisol levels between individuals that recovered in visual contact with their own social group and those in visual contact with a novel but non-aggressive social group. Collectively, our results suggest that affiliation and familiarity per se do not mediate social buffering in this group-living cichlid, and the behavioural and physiological mechanisms responsible for social buffering may vary across vertebrates.
Assuntos
Ciclídeos/fisiologia , Comportamento Social , Animais , Feminino , Masculino , Ocitocina/análogos & derivados , Ocitocina/metabolismo , Vasotocina/metabolismoRESUMO
The nonapeptide arginine vasotocin (AVT) regulates osmotic balance in teleost fishes, but its mechanisms of action are not fully understood. Recently, it was discovered that nonapeptide receptors in teleost fishes are differentiated into two V1a-type, several V2-type, and two isotocin (IT) receptors, but it remains unclear which receptors mediate AVT's effects on gill osmoregulation. Here, we examined the role of nonapeptide receptors in the gill of the euryhaline Amargosa pupfish (Cyprinodon nevadensis amargosae) during osmotic acclimation. Transcripts for the teleost V1a-type receptor v1a2 were upregulated over fourfold in gill 24 h after transferring pupfish from 7.5 ppt to seawater (35 ppt) or hypersaline (55 ppt) conditions and downregulated after transfer to freshwater (0.3 ppt). Gill transcripts for the nonapeptide degradation enzyme leucyl-cystinyl aminopeptidase (LNPEP) also increased in fish acclimating to 35 ppt. To test whether the effects of AVT on the gill might be mediated by a V1a-type receptor, we administered AVT or a V1-type receptor antagonist (Manning compound) intraperitoneally to pupfish before transfer to 0.4 ppt or 35 ppt. Pupfish transferred to 35 ppt exhibited elevated gill mRNA abundance for cystic fibrosis transmembrane conductance regulator (cftr), but that upregulation diminished under V1-receptor inhibition. AVT inhibited the increase in gill Na+/Cl- cotransporter 2 (ncc2) transcript abundance that occurs following transfer to hypoosmotic environments, whereas V1-type receptor antagonism increased ncc2 mRNAs even without a change in salinity. These findings indicate that AVT acts via a V1-type receptor to regulate gill Cl- transport by inhibiting Cl- uptake and facilitating Cl- secretion during seawater acclimation.
Assuntos
Proteínas de Peixes/metabolismo , Brânquias/metabolismo , Peixes Listrados/metabolismo , Osmorregulação , Receptores de Vasopressinas/metabolismo , Salinidade , Tolerância ao Sal , Vasotocina/metabolismo , Animais , Regulador de Condutância Transmembrana em Fibrose Cística/genética , Regulador de Condutância Transmembrana em Fibrose Cística/metabolismo , Cistinil Aminopeptidase/genética , Cistinil Aminopeptidase/metabolismo , Feminino , Proteínas de Peixes/genética , Peixes Listrados/genética , Masculino , Ocitocina/análogos & derivados , Ocitocina/metabolismo , Receptores de Vasopressinas/genética , Água do Mar , Transdução de Sinais , Membro 1 da Família 12 de Carreador de Soluto/genética , Membro 1 da Família 12 de Carreador de Soluto/metabolismo , Regulação para CimaRESUMO
The hypothalamic neuropeptides arginine vasopressin (AVP) and oxytocin (OT) modulate social behavior across a wide variety of species. However, the role of arginine vasotocin (AVT) and isotocin (IT, the teleost homologs of AVP and OT) in regulating biparental care especially in the context of monogamy is not well studied. Here, using high-performance liquid chromatography (HPLC), we investigated how bioactive whole brain AVT and IT neuropeptide levels vary in relation to the phase of the breeding cycle and sex, in a monogamous biparental cichlid fish, Neolamprologus caudopunctatus. Since non-caring individuals of this species readily cannibalize eggs, but caring parents never do, we further investigated whether there might be changes in AVT/IT whole brain levels that correspond to the transition from a non-breeding, egg cannibal to an egg caring parent. We found that AVT levels were higher in females than in males and that AVT levels were highest when the need to defend the young was greatest. Breeding pairs that had a strong pair-bond and a higher frequency of nest care had the highest levels of AVT, whereas individuals that spent little time close to their breeding partner, displayed aggression towards their partner and neglected their nest duties (signs of a weak pair bond), had lower whole brain AVT levels. Isotocin (IT) levels did not differ between sexes and we did not detect any variation across the breeding cycle, with pair-bonding scores or with parental behavior. Our results show that whole brain AVT levels are linked to the breeding cycle, nest maintenance and pair-bonding score in this species. Furthermore, our study highlights species and sex-specific nonapeptides patterns in bi-parental caring fish and contributes to the increasing knowledge on neuroendocrinological mechanisms underlying parental care.
Assuntos
Arginina Vasopressina/metabolismo , Ciclídeos/fisiologia , Comportamento de Nidação/fisiologia , Ocitocina/metabolismo , Agressão/fisiologia , Animais , Encéfalo/metabolismo , Feminino , Masculino , Comportamento Materno/fisiologia , Ligação do Par , Comportamento Paterno/fisiologiaRESUMO
The nonapeptide oxytocin (and its fish homolog isotocin (IT)) is an evolutionarily-conserved hormone associated with social behaviors across most vertebrate taxa. Oxytocin has traditionally been regarded as a prosocial hormone, but studies on social cognition in mammalian models suggest it may play a more nuanced role in modulating social discrimination based on social salience and stimulus valence. Here we test IT and its role in regulating female social decision-making and anxiety behaviors in a live-bearing fish with a male coercive mating system. Gambusia affinis males engage in a forced mating strategy, with frequent harassment and attempted copulatory thrusts directed towards unwilling females. Exogenous IT produced anxiolytic responses in female G. affinis that altered exploration (time in center of tank) but not time in dark vs. light regions of the tank. Exogenous IT also produced context-specific changes in social tendency: IT-treated G. affinis females spent less time associating with males while association time with conspecific females was not altered. Further, while overall activity levels were not changed by IT treatment, the amount of social behaviors IT-treated females directed towards males, but not females, was reduced. Our results support the social salience hypothesis of oxytocin action in a teleost and suggest that oxytocin's critical input into social cognitive processing is conserved across vertebrate taxa.
Assuntos
Agressão , Aprendizagem da Esquiva/efeitos dos fármacos , Ciprinodontiformes/fisiologia , Ocitocina/análogos & derivados , Comportamento Sexual Animal , Agressão/efeitos dos fármacos , Agressão/fisiologia , Animais , Ansiedade/etiologia , Coerção , Cognição/efeitos dos fármacos , Feminino , Masculino , Ocitocina/farmacologia , Ocitocina/fisiologia , Poecilia/fisiologia , Comportamento Sexual Animal/efeitos dos fármacos , Comportamento Sexual Animal/fisiologia , Comportamento SocialRESUMO
Maintenance of internal ionic and acid-base homeostasis is critical for survival in all biological systems. Similar to mammals, aquatic fishes have developed sophisticated homeostatic mechanisms to mitigate metabolic or environmental disruptions in ionic and acid-base status of systemic body fluids via hormone-controlled transport of ions or acid equivalents. The present review summarizes newly discovered actions of several hormones in zebrafish (Danio rerio) and medaka (Oryzias latipes) that have greatly contributed to our overall understanding of ionic/acid-base regulation. For example, isotocin and cortisol were reported to enhance transport of various ions by stimulating the proliferation and/or differentiation of ionocyte progenitors. Meanwhile, stanniocalcin-1, a well-documented hypocalcemic hormone, was found to suppress ionocyte differentiation and thus downregulate secretion of H+ and uptake of Na+ and Cl-. Estrogen-related receptor and calcitonin gene-related peptide also regulate the differentiation of certain types of ionocytes to either stimulate or suppress H+ secretion and Cl- uptake. On the other hand, endothelin and insulin-like growth factor 1 activate the respective secretion of H+ and Na+/Cl through fast actions. These new findings enhance our understanding of how hormones regulate fish ionic and acid-base regulation while further providing new insights into vertebrate evolution, mammalian endocrinology and human disease-related therapeutics.
Assuntos
Equilíbrio Ácido-Base , Hormônios/metabolismo , Oryzias/metabolismo , Osmorregulação , Peixe-Zebra/metabolismo , Animais , Modelos BiológicosRESUMO
Fathering behavior is critical for offspring survival in many species across diverse taxa, but our understanding of the neuroendocrine mechanisms regulating paternal care is limited in part because of the few primarily paternal species among the common animal models. However, many teleosts display primarily paternal care, and among the teleosts, anemonefish species are particularly well suited for isolating molecular mechanisms of fathering as they perform parental care in isolation of many other typically competing behaviors such as territorial defense and nest building. The goal of this study was to determine the extent to which whole brain gene expression levels of isotocin receptors, arginine vasotocin receptors, and aromatase as well as circulating levels of the bioactive sex steroid hormones estradiol (E2) and 11-ketotestosterone (11KT) vary in association with parenting behavior in Amphiprion ocellaris. Brain aromatase and IT receptor gene expression were higher in both males and females that were parenting versus not. IT receptor expression was overall higher in males than females, which we interpret is a reflection of the greater parental effort that males display. Aromatase was overall higher in females than males, which we conclude is related to the higher circulating E2, which crosses into the brain and increases aromatase transcription. Results suggest both aromatase and IT receptors are dynamically upregulated in the brains of A. ocellaris males and females to support high levels of parental effort.
Assuntos
Aromatase/genética , Encéfalo/metabolismo , Comportamento de Nidação/fisiologia , Perciformes/genética , Receptores de Ocitocina/genética , Animais , Aromatase/metabolismo , Encéfalo/enzimologia , Estradiol/sangue , Feminino , Regulação da Expressão Gênica , Masculino , Comportamento Materno/fisiologia , Comportamento Paterno/fisiologia , Perciformes/metabolismo , Receptores de Ocitocina/metabolismo , Receptores de Vasopressinas/genética , Receptores de Vasopressinas/metabolismo , Anêmonas-do-Mar , Testosterona/análogos & derivados , Testosterona/sangueRESUMO
In the present study, in vitro effects of synthetic vasotocin (VT), isotocin (4Ser, 8Ile- oxytocin; ITb) and the recently cloned IT gene paralog product (8Val-Isotocin, ITa) were studied on the expression of pituitary gonadotropin (GtH) subunit mRNA levels. In male pituitaries of early (preparatory phase) and late (prespawning phase) recrudescing catfish, Heteropneustes fossilis, VT (10â¯nM, 100â¯nM and 1000â¯nM) stimulated fshß expression dose-dependently. But in females, the dose-dependent effect was found only in the preparatory phase. In males, VT stimulated lhß expression only at higher doses. In females, VT produced a significant dose-dependent increase of the lhß expression only in the prespawning phase. VT stimulated the expression of gpα, dose-dependently in the preparatory phase in males and in the prespawning phase in females. The incubation of the pituitaries with ITb did not alter the fshß expression in either sex in both preparatory and prespawning phases. In males, ITb stimulated the expression of lhß and gpα only at the highest concentration (1000â¯nM) in both phases. In females, ITb stimulated both lhß and gpα expression only at 1000â¯nM in the preparatory phase and dose-dependently in the prespawning phase. The incubation of the pituitaries with ITa produced effects similar to ITb on the expression of fshß, lhß, and gpα. The results show that the basic peptide VT modulates both fshß and lhß expressions, which are influenced by the sex and reproductive stage. The neutral peptide ITA/ITb exerts an insignificant effect on the fshß expression regardless of sex or season. Both VT and ITa/ITb elicit a significant effect on the lhß expression in late recrudescent phase especially in females.
Assuntos
Peixes-Gato , Gonadotropinas Hipofisárias/genética , Hormônios Neuro-Hipofisários/farmacologia , Reprodução/efeitos dos fármacos , Maturidade Sexual/efeitos dos fármacos , Animais , Peixes-Gato/genética , Peixes-Gato/crescimento & desenvolvimento , Peixes-Gato/metabolismo , Feminino , Subunidade beta do Hormônio Folículoestimulante/genética , Subunidade beta do Hormônio Folículoestimulante/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Gonadotropinas Hipofisárias/metabolismo , Técnicas In Vitro , Hormônio Luteinizante Subunidade beta/genética , Hormônio Luteinizante Subunidade beta/metabolismo , Masculino , Ocitocina/análogos & derivados , Ocitocina/farmacologia , Hipófise/efeitos dos fármacos , Hipófise/metabolismo , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Reprodução/genética , Estações do Ano , Caracteres Sexuais , Maturidade Sexual/genética , Vasotocina/farmacologiaRESUMO
Gilthead sea bream, Sparus aurata L., is an important fish species for the Mediterranean aquaculture and is considered a good model for studying the osmoregulatory process, due to its capacity to cope with great changes in environmental salinity (5-60). Our group studied the osmoregulatory role of different endocrine systems in this species, focusing on the vasotocinergic and isotocinergic systems over several years. For this purpose, the cDNAs coding for pro-vasotocin (pro-vt), pro-isotocin (pro-it), two arginine vasotocin (AVT) receptors (avtr v1a2- and v2-types) and one IT receptor (itr) were cloned. Acclimation to different environmental salinities induced a direct lineal relationship between plasma AVT levels and salinity, with no changes in plasma IT values. In addition, higher values in vasotocinergic, isotocinergic and stress pathways (pro-vt and pro-it gene expression, AVT and IT storage and plasma cortisol levels) in both hypo- and/or hyper-osmotic transfers, suggest an interaction between cortisol and AVT/IT pathways. Moreover, gene expression of specific receptors, as well as the use of different in vitro techniques, demonstrated an important osmoregulatory orchestration in different organs. In addition, individuals intraperitoneally injected with AVT and transferred to different environmental salinities enhanced plasma cortisol levels and/or gill Na+, K+-ATPase activity. These effects could be related to the energy repartitioning process occurring during osmotic adaptation of S. aurata to extreme environmental salinities, which could be mediated not only by plasma cortisol but also by AVT. Finally, our results indicated a very important role of the vasotocinergic and/or isotocinergic systems in both osmoregulatory and non-osmoregulatory organs.
Assuntos
Receptores de Vasopressinas/metabolismo , Dourada , Vasotocina/metabolismo , Animais , OsmorregulaçãoRESUMO
The influence of chronic stress, induced by food deprivation (FD) and/or high stocking density (HSD), was assessed on stress, vasotocinergic and isotocinergic pathways of the gilthead sea bream (Sparus aurata). Fish were randomly assigned to one of the following treatments: (1) fed at low stocking density (LSD-F; 5kg·m-3); (2) fed at high stocking density (HSD-F, 40kg·m-3); (3) food-deprived at LSD (LSD-FD); and (4) food-deprived at HSD (HSD-FD). After 21days, samples from plasma, liver, hypothalamus, pituitary and head-kidney were collected. Both stressors (FD and HSD) induced a chronic stress situation, as indicated by the elevated cortisol levels, the enhancement in corticotrophin releasing hormone (crh) expression and the down-regulation in corticotrophin releasing hormone binding protein (crhbp) expression. Changes in plasma and liver metabolites confirmed a metabolic adjustment to cope with energy demand imposed by stressors. Changes in avt and it gene expression, as well as in their specific receptors (avtrv1a, avtrv2 and itr) at central (hypothalamus and pituitary) and peripheral (liver and head-kidney) levels, showed that vasotocinergic and isotocinergic pathways are involved in physiological changes induced by FD or HSD, suggesting that different stressors are handled through different stress pathways in S. aurata.
Assuntos
Fenômenos Fisiológicos da Nutrição Animal , Proteínas de Peixes/metabolismo , Modelos Neurológicos , Ocitocina/análogos & derivados , Dourada/fisiologia , Estresse Fisiológico , Vasotocina/metabolismo , Animais , Biomarcadores/sangue , Biomarcadores/metabolismo , Restrição Calórica/efeitos adversos , Aglomeração , Proteínas de Peixes/sangue , Proteínas de Peixes/genética , Regulação da Expressão Gênica no Desenvolvimento , Rim Cefálico/crescimento & desenvolvimento , Rim Cefálico/inervação , Rim Cefálico/metabolismo , Hipotálamo/crescimento & desenvolvimento , Hipotálamo/metabolismo , Fígado/crescimento & desenvolvimento , Fígado/metabolismo , Masculino , Neurônios/metabolismo , Ocitocina/sangue , Ocitocina/metabolismo , Hipófise/crescimento & desenvolvimento , Hipófise/inervação , Hipófise/metabolismo , Distribuição Aleatória , Dourada/sangue , Dourada/crescimento & desenvolvimento , Vasotocina/sangueRESUMO
Most wrasses are protogynous species that swim to feed, reproduce during the daytime, and bury themselves under the sandy bottom at night. In temperate and subtropical wrasses, low temperature influences emergence from the sandy bottom in the morning, and induces a hibernation-like state in winter. We cloned and characterized the prohormone complementary DNAs (cDNAs) of arginine vasotocin (AVT) and isotocin (IT) in a temperate wrasse (Halichoeres tenuispinis) and examined the effects of day/night and temperature on their expression in the diencephalon, because these neurohypophysial peptides are related to the sex behavior of wrasses. The full-length cDNAs of pro-AVT and pro-IT were 938 base pairs (154 amino acids) and 759 base pairs (156 amino acids) in length, respectively. Both pro-peptides contained a signal sequence followed by the respective hormones and neurophysin connected by a Gly-Lys-Arg bridge. Reverse-transcription polymerase chain reaction (RT-PCR) revealed that pro-AVT mRNA expression was specifically observed in the diencephalon, whereas pro-IT mRNA expression was seen in the whole brain. Quantitative RT-PCR revealed that the mRNA abundance of pro-AVT and pro-IT was higher at midday (zeitgeber time 6; ZT6) than at midnight (ZT18) under 12 h light and 12 h darkness (LD 12:12) conditions, but not under constant light. Intraperitoneal injection of melatonin decreased the mRNA abundance of pro-AVT, but not of pro-IT. When fish were reared under LD 12:12 conditions at 25, 20, and 15 °C, day high and night low mRNA expressions of pro-AVT and pro-IT were maintained. A field survey revealed seasonal variation in the number of swimming fish at observatory sites; many fish emerged from the sandy bottom in summer, but not in winter, suggesting a hibernation-like state under the sandy bottom under low temperature conditions. We conclude that the day-night fluctuation of pro-AVT and pro-IT mRNA abundance in the brain is not affected by temperature and repeated under the sandy bottom in winter.
Assuntos
Diencéfalo/metabolismo , Peixes/genética , Melatonina/farmacologia , Ocitocina/análogos & derivados , Temperatura , Vasotocina/genética , Animais , Diencéfalo/efeitos dos fármacos , Ocitocina/genética , Periodicidade , RNA Mensageiro/metabolismoRESUMO
The nonapeptides isotocin (IT) and arginine vasotocin (AVT), along with their mammalian homologs oxytocin and arginine vasopressin, are well known regulators of social behaviors across vertebrate taxa. However, little is known about their involvement in paternal care. Here, we measured the effect of an IT and an AVT V1a receptor antagonist on paternal behaviors in the primarily paternal teleost Amphiprion ocellaris. We also measured the effect of the IT receptor antagonist on aggression in dyadic contests between two non-reproductive fish to assess specificity of the effect on paternal behaviors. Individual differences in levels of paternal behaviors (nips, fanning the eggs, and proportion of the time in the nest) were consistent across spawning cycles when no treatments were administered. The IT receptor antagonist severely reduced paternal behaviors but had no effect on aggression, whereas the AVT V1a receptor antagonist increased paternal behaviors. These results support the idea that IT signaling is crucial for the expression of paternal behavior in A. ocellaris. Based on a previous study showing that the AVT V1a antagonist decreases aggression in dyadic contests, we hypothesize that the antagonist enhances paternal behavior indirectly by reducing vigilance and aggression, thereby alleviating effort directed towards other competing behaviors and allowing for the increased expression of paternal behaviors.
Assuntos
Agressão/efeitos dos fármacos , Antagonistas de Hormônios/farmacologia , Comportamento Paterno/efeitos dos fármacos , Perciformes/fisiologia , Animais , Arginina Vasopressina/metabolismo , Feminino , Masculino , Comportamento de Nidação/efeitos dos fármacos , Ocitocina/análogos & derivados , Ocitocina/metabolismo , Perciformes/metabolismo , Receptores de Vasopressinas/metabolismo , Comportamento Social , Vasotocina/metabolismoRESUMO
The vasotocin/vasopressin and isotocin/mesotocin/oxytocin family of nonapeptides regulate social behaviors and physiological functions associated with reproductive physiology and osmotic balance. While experimental and correlative studies provide evidence for these nonapeptides as modulators of behavior across all classes of vertebrates, mechanisms for nonapeptide inactivation in regulating these functions have been largely overlooked. Leucyl-cystinyl aminopeptidase (LNPEP) - also known as vasopressinase, oxytocinase, placental leucine aminopeptidase (P-LAP), and insulin-regulated aminopeptidase (IRAP) - is a membrane-bound zinc-dependent metalloexopeptidase enzyme that inactivates vasopressin, oxytocin, and select other cyclic polypeptides. In humans, LNPEP plays a key role in the clearance of oxytocin during pregnancy. However, the evolutionary diversity, expression distribution, and functional roles of LNPEP remain unresolved for other vertebrates. Here, we isolated and sequenced a full-length cDNA encoding a LNPEP-like polypeptide of 1033 amino acids from the ovarian tissue of Amargosa pupfish, Cyprinodon nevadensis. This deduced polypeptide exhibited high amino acid identity to human LNPEP both in the protein's active domain that includes the peptide binding site and zinc cofactor binding motif (53.1% identity), and in an intracellular region that distinguishes LNPEP from other aminopeptidases (70.3% identity). Transcripts encoding this LNPEP enzyme (lnpep) were detected at highest relative abundance in the gonads, hypothalamus, forebrain, optic tectum, gill and skeletal muscle of adult pupfish. Further evaluation of lnpep transcript abundance in the brain of sexually-mature pupfish revealed that lnpep mRNAs were elevated in the hypothalamus of socially subordinate females and males, and at lower abundance in the telencephalon of socially dominant males compared to dominant females. These findings provide evidence of an association between behavioral social status and hypothalamic lnpep transcript abundance and suggest that variation in the rate of VT/IT peptide inactivation by LNPEP may be a contributing component in the mechanism whereby nonapeptides regulate social behavior.