RESUMO
Vasopressin (AVP) plays an important role in anxiety-related and social behaviors. Single-prolonged stress (SPS) has been established as an animal acute severe stress model and has been shown to induce a lower adrenocorticotropic hormone (ACTH) response upon cortisol challenge. Here, we show results from immunoassays for AVP, ACTH, and corticosterone (CORT), and in situ hybridizations for AVP mRNA performed 7 days after SPS exposure. Immunofluorescence for AVP was also performed during the 7-day period following SPS exposure and after an additional forced swimming stress paradigm. We observed that the plasma concentrations of AVP, ACTH, and CORT were not altered by SPS; ACTH content in the pituitary and AVP mRNA expression in the supraoptic nucleus (SON) were significantly reduced by SPS. During the 7-day period following SPS, the intensity of immunoreactivity, the size of the soma, and the immunoreactive optical density of the dendrites of AVP neurons in the SON all increased. An apparent reduction in the intensity of AVP immunoreactivity was observed in the SON at 4 h after additional stress. Additional forced swimming led to a rapid increase in the dendritic AVP content only in the controls and not in the SPS-treated rats. These findings suggest that AVP is a potential biomarker for past exposure to severe stress and that alterations in AVP may affect the development of pathogenesis in stress-related disorders.
Assuntos
Neurônios/metabolismo , Estresse Psicológico/metabolismo , Estresse Psicológico/patologia , Núcleo Supraóptico/patologia , Vasopressinas/metabolismo , Hormônio Adrenocorticotrópico/metabolismo , Análise de Variância , Animais , Comportamento Animal , Corticosterona/metabolismo , Dendritos/metabolismo , Dendritos/patologia , Modelos Animais de Doenças , Regulação da Expressão Gênica/fisiologia , Masculino , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Natação , Fatores de Tempo , Vasopressinas/genéticaRESUMO
The effects of intraperitoneal (i.p.) administration of 2-buten-4-olide (2-B4O), an endogenous sugar acid, on the hypothalamo-adenohypophysial system were examined in Lewis rats that were normal and in adjuvant-induced arthritic (AA) rats. In comparison with vehicle-treated rats, the plasma corticosterone and c-fos mRNA levels in the paraventricular nucleus (PVN) of normal rats increased significantly after i.p. administration of 2-B4O. Dual immunostaining revealed that almost all corticotrophin-releasing factor (CRF)-immunopositive neurones in the parvocellular division of the PVN exhibited Fos-like immunoreactivity (LI) 120 min after i.p. administration of 2-B4O (100 mg/kg). In the AA rats, repeated i.p. administration of 2-B4O (100 mg/kg) after immunisation significantly suppressed the expression of clinical symptoms and significantly increased plasma concentrations of corticosterone. Further, repeated i.p. administration of 2-B4O significantly increased CRF mRNA levels in the PVN and pro-opiomelanocortin mRNA levels in the anterior pituitary; however, they did not change arginine vasopressin mRNA levels in the parvocellular division of the PVN. These results suggest that i.p. administration of 2-B4O activates the hypothalamo-pituitary-adrenal (HPA) axis via the activation of CRF neurones in the PVN, and the activation of the HPA axis by i.p. administration of 2-B4O may be associated with the inhibition of AA in rats.
Assuntos
4-Butirolactona/análogos & derivados , Artrite Experimental , Sistema Hipotálamo-Hipofisário/efeitos dos fármacos , Sistema Hipófise-Suprarrenal/efeitos dos fármacos , 4-Butirolactona/administração & dosagem , 4-Butirolactona/farmacologia , Adjuvantes Imunológicos , Animais , Depressores do Apetite/farmacologia , Arginina Vasopressina/metabolismo , Artrite Experimental/sangue , Artrite Experimental/metabolismo , Corticosterona/sangue , Hormônio Liberador da Corticotropina/metabolismo , Sistema Hipotálamo-Hipofisário/metabolismo , Injeções Intraperitoneais , Masculino , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ratos , Ratos Endogâmicos LewRESUMO
Dasatinib treatment markedly increases the number of large granular lymphocytes (LGLs) in a proportion of Ph+ leukemia patients, which associates with a better prognosis. The lymphocytosis is predominantly observed in cytomegalovirus (CMV)-seropositive patients, yet detectable CMV reactivation exists only in a small fraction of patients. Thus, etiology of the lymphocytosis still remains unclear. Here, we identified NK cells as the dominant LGLs expanding in dasatinib-treated patients, and applied principal component analysis (PCA) to an extensive panel of NK cell markers to explore underlying factors in NK cell activation. PCA displayed phenotypic divergence of NK cells that reflects CMV-associated differentiation and genetic differences, and the divergence was markedly augmented in CMV-seropositive dasatinib-treated patients. Notably, the CMV-associated highly differentiated status of NK cells was already observed at leukemia diagnosis, and was further enhanced after starting dasatinib in virtually all CMV-seropositive patients. Thus, the extensive characterization of NK cells by PCA strongly suggests that CMV is an essential factor in the NK cell activation, which progresses stepwise during leukemia and subsequent dasatinib treatment most likely by subclinical CMV reactivation. This study provides a rationale for the exploitation of CMV-associated NK cell activation for treatment of leukemias.
Assuntos
Citomegalovirus , Dasatinibe/uso terapêutico , Células Matadoras Naturais/imunologia , Leucemia Mielogênica Crônica BCR-ABL Positiva/imunologia , Análise de Componente Principal , Humanos , Células Matadoras Naturais/microbiologia , Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Ativação ViralRESUMO
The effects of i.c.v. administration of prolactin-releasing peptide on neurons in the paraventricular nucleus of rats and plasma corticosterone levels were examined by measuring changes in Fos-like immunoreactivity, c-fos mRNA using in situ hybridization histochemistry, and plasma corticosterone using a specific radioimmunoassay. Approximately 80% of corticotropin-releasing hormone immunoreactive cells exhibited Fos-like immunoreactivity in the parvocellular division of the paraventricular nucleus 90 min after i.c.v. administration of prolactin-releasing peptide. The greatest induction of the c-fos mRNA expression in the paraventricular nucleus was observed 30 min after administration of prolactin-releasing peptide, and occurred in a dose-related manner. Plasma corticosterone levels were also significantly increased 30 min after administration of prolactin-releasing peptide. Next, the effects of restraint stress, nociceptive stimulus and acute inflammatory stress on the expression of the prolactin-releasing peptide mRNA in the dorsomedial hypothalamic nucleus, nucleus of the solitary tract and ventrolateral medulla were examined using in situ hybridization histochemistry for prolactin-releasing peptide mRNA. Restraint stress and acute inflammatory stress upregulated the prolactin-releasing peptide mRNA expression in the nucleus of the solitary tract and ventrolateral medulla. Nociceptive stimulus upregulated the prolactin-releasing peptide mRNA expression in the ventrolateral medulla. Finally, we observed that pretreatment (i.c.v. administration) with an anti-prolactin-releasing peptide antibody significantly attenuated nociceptive stimulus-induced c-fos mRNA expression in the paraventricular nucleus. These results suggest that prolactin-releasing peptide is a potent and important mediator of the stress response in the brain through the hypothalamic paraventricular nucleus.
Assuntos
Encéfalo/fisiopatologia , Hormônios Hipotalâmicos/fisiologia , Neurônios/metabolismo , Neuropeptídeos/fisiologia , Núcleo Hipotalâmico Paraventricular/fisiologia , Estresse Fisiológico/metabolismo , Análise de Variância , Animais , Anti-Inflamatórios não Esteroides/administração & dosagem , Encéfalo/efeitos dos fármacos , Corticosterona/sangue , Hormônio Liberador da Corticotropina/metabolismo , Relação Dose-Resposta a Droga , Hormônios Hipotalâmicos/imunologia , Imunoglobulina G/administração & dosagem , Indometacina/administração & dosagem , Lipopolissacarídeos/toxicidade , Masculino , Neuropeptídeos/imunologia , Medição da Dor/métodos , Núcleo Hipotalâmico Paraventricular/citologia , Núcleo Hipotalâmico Paraventricular/efeitos dos fármacos , Hormônio Liberador de Prolactina , Proteínas Proto-Oncogênicas c-fyn/genética , Proteínas Proto-Oncogênicas c-fyn/metabolismo , Radioimunoensaio/métodos , Ratos , Ratos Wistar , Restrição Física/métodos , Estresse Fisiológico/etiologiaRESUMO
We examined the effects of chronic salt loading on the hypothalamic expressions of the enhanced green fluorescent protein (eGFP), arginine vasopressin (AVP) and oxytocin (OXT) genes in AVP-eGFP transgenic rats that expressed eGFP in the hypothalamic AVP-containing neurones. In these rats, salt loading for 5 days caused a marked increase of the eGFP fluorescence in the magnocellular divisions of the paraventricular nucleus (PVN), the supraoptic nucleus (SON) and the internal layer of the median eminence. Expression of the eGFP gene was increased seven- to eight-fold in the PVN and SON of salt-loaded rats in comparison with euhydrated rats. By contrast, none of these changes were observed in the suprachiasmatic nucleus. The expression of the AVP and OXT genes was increased 1.5- to two-fold in the PVN and SON of salt-loaded nontransgenic (control) and transgenic rats. There were no differences in the expression levels of the AVP and OXT genes in the PVN and SON between nontransgenic (control) and transgenic animals under normal conditions and after salt loading. In the posterior pituitary gland, the intensity of the eGFP fluorescence did not change after salt loading for 5 days, but increased after 10 days of salt loading. Upon salt loading, significant increases in the plasma AVP concentrations, plasma osmolality and plasma Na+ were observed. Furthermore, there were no significant differences in changes of water intake, food intake, urine volume, urine osmolality, urine Na+ concentrations, and the body weights in both models under normal or salt-loaded conditions. Our results show that the response of the AVP-eGFP fusion gene to chronic salt loading is exaggerated, and humoral responses such as AVP and OXT and the body fluid homeostasis are maintained in AVP-eGFP transgenic rats. The AVP-eGFP transgenic rat gives us a new opportunity to study the dynamics of the AVP system in vivo.
Assuntos
Arginina Vasopressina/biossíntese , Arginina Vasopressina/genética , Homeostase/fisiologia , Cloreto de Sódio/farmacologia , Equilíbrio Hidroeletrolítico/fisiologia , Animais , Animais Geneticamente Modificados , Arginina Vasopressina/fisiologia , Peptídeo Semelhante a Galanina/metabolismo , Proteínas de Fluorescência Verde/biossíntese , Proteínas de Fluorescência Verde/genética , Hibridização In Situ , Masculino , Microscopia de Fluorescência , Concentração Osmolar , Ocitocina/biossíntese , Ocitocina/sangue , Ocitocina/fisiologia , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Radioimunoensaio , Ratos , Ratos Wistar , Núcleo Supraquiasmático/efeitos dos fármacos , Núcleo Supraquiasmático/metabolismo , Equilíbrio Hidroeletrolítico/efeitos dos fármacosRESUMO
Oxytocin (OXT)-containing neurosecretory cells in the parvocellular divisions of the paraventricular nucleus (PVN), which project to the medulla and spinal cord, are involved in various physiological functions, such as sensory modulation and autonomic processes. In the present study, we examined OXT expression in the hypothalamo-spinal pathway, as well as the hypothalamo-neurohypophysial system, which includes the magnocellular neurosecretory cells in the PVN and the supraoptic nucleus (SON), after s.c. injection of saline or formalin into the hindpaws of transgenic rats that express the OXT and monomeric red fluorescent protein 1 (mRFP1) fusion gene. (i) The numbers of OXT-mRFP1 neurones that expressed Fos-like immunoreactivity (-IR) and OXT-mRFP1 intensity were increased significantly in the magnocellular/parvocellular PVN and SON after s.c. injection of formalin. (ii) OXT-mRFP1 neurones in the anterior parvocellular PVN, which may project to the dorsal horn of the spinal cord, were activated by s.c. injection of formalin, as indicated by a significant increases of Fos-IR and mRFP1 intensity intensity. (iii) Formalin injection caused a significant transient increase in plasma OXT. (iv) OXT, mRFP1 and corticotrophin-releasing hormone mRNAs in the PVN were significantly increased after s.c. injection of formalin. (v) An intrathecal injection of OXT-saporin induced hypersensitivity in conscious rats. Taken together, these results suggest that the hypothalamo-neurohypophysial/-spinal OXTergic pathways may be involved in acute nociceptive responses in rats.
Assuntos
Hiperalgesia/induzido quimicamente , Hiperalgesia/fisiopatologia , Hipotálamo/metabolismo , Ocitocina/fisiologia , Neuro-Hipófise/metabolismo , Animais , Hormônio Liberador da Corticotropina/biossíntese , Formaldeído , Injeções Espinhais , Proteínas Luminescentes/genética , Masculino , Neurônios/metabolismo , Ocitocina/administração & dosagem , Ocitocina/análogos & derivados , Ocitocina/biossíntese , Ocitocina/sangue , Ocitocina/farmacologia , Medição da Dor , Núcleo Hipotalâmico Paraventricular/metabolismo , Núcleo Hipotalâmico Paraventricular/fisiologia , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ratos , Ratos Transgênicos , Proteínas Inativadoras de Ribossomos Tipo 1/administração & dosagem , Proteínas Inativadoras de Ribossomos Tipo 1/farmacologia , Saporinas , Núcleo Supraóptico/metabolismo , Núcleo Supraóptico/fisiologia , Proteína Vermelha FluorescenteRESUMO
How does a neuron, challenged by an increase in synaptic input, display a response that is independent of the initial level of activity? Here we show that both oxytocin and vasopressin cells in the supraoptic nucleus of normal rats respond to intravenous infusions of hypertonic saline with gradual, linear increases in discharge rate. In hyponatremic rats, oxytocin and vasopressin cells also responded linearly to intravenous infusions of hypertonic saline but with much lower slopes. The linearity of response was surprising, given both the expected nonlinearity of neuronal behavior and the nonlinearity of the oxytocin secretory response to such infusions. We show that a simple computational model can reproduce these responses well, but only if it is assumed that hypertonic infusions coactivate excitatory and inhibitory synaptic inputs. This hypothesis was tested first by applying the GABA(A) antagonist bicuculline to the dendritic zone of the supraoptic nucleus by microdialysis. During local blockade of GABA inputs, the response of oxytocin cells to hypertonic infusion was greatly enhanced. We then went on to directly measure GABA release in the supraoptic nucleus during hypertonic infusion, confirming the predicted rise. Together, the results suggest that hypertonic infusions lead to coactivation of excitatory and inhibitory inputs and that this coactivation may confer appropriate characteristics on the output behavior of oxytocin cells. The nonlinearity of oxytocin secretion that accompanies the linear increase in oxytocin cell firing rate reflects frequency-facilitation of stimulus-secretion coupling at the neurohypophysis.
Assuntos
Simulação por Computador , Modelos Neurológicos , Inibição Neural/fisiologia , Neurônios/fisiologia , Núcleo Supraóptico/fisiologia , Animais , Bicuculina/administração & dosagem , Desamino Arginina Vasopressina , Eletrofisiologia , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Antagonistas GABAérgicos/administração & dosagem , Hiponatremia/sangue , Hiponatremia/induzido quimicamente , Infusões Intravenosas , Masculino , Microdiálise , Neurônios/classificação , Neurônios/efeitos dos fármacos , Concentração Osmolar , Ocitocina/sangue , Ratos , Ratos Sprague-Dawley , Ratos Wistar , Solução Salina Hipertônica/administração & dosagem , Sódio/sangue , Estimulação Química , Núcleo Supraóptico/citologia , Núcleo Supraóptico/efeitos dos fármacos , Vasopressinas/agonistas , Vasopressinas/metabolismo , Ácido gama-Aminobutírico/metabolismoRESUMO
In the present study, we investigated the effects of the cannabinoid receptor agonist CP55,940 on excitatory and inhibitory synaptic transmission in the rat supraoptic nucleus. Whole-cell patch clamp recordings were performed on supraoptic neurones in in vitro brain slice preparations. CP55,940 significantly reduced the frequency of spontaneous excitatory and inhibitory postsynaptic currents in a concentration-dependent manner. These changes were potently reversed by the CB1 receptor antagonist AM251. The results indicate that cannabinoids modulate the activity of magnocellular neurosecretory neurones by presynaptic inhibition of both excitatory and inhibitory synaptic transmission.
Assuntos
Canabinoides/farmacologia , Núcleo Supraóptico/fisiologia , Sinapses/efeitos dos fármacos , Anestésicos Locais/farmacologia , Animais , Cicloexanóis/farmacologia , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Técnicas In Vitro , Masculino , Potenciais da Membrana/efeitos dos fármacos , Neurônios Aferentes/efeitos dos fármacos , Técnicas de Patch-Clamp , Piperidinas/farmacologia , Pirazóis/farmacologia , Ratos , Ratos Wistar , Receptor CB1 de Canabinoide/antagonistas & inibidores , Receptores Pré-Sinápticos/efeitos dos fármacos , Núcleo Supraóptico/efeitos dos fármacos , Tetrodotoxina/farmacologiaRESUMO
CONTEXT: A chemical scenario was proposed for photon-dominated regions (PDRs) according to which UV photons from nearby stars lead to the evaporation of very small grains (VSGs) and the production of gas-phase polycyclic aromatic hydrocarbons (PAHs). AIMS: Our goal is to achieve better insight into the composition and evolution of evaporating very small grains (eVSGs) and PAHs through analyzing the infrared (IR) aliphatic and aromatic emission bands. METHODS: We combined spectro-imagery in the near- and mid-IR to study the spatial evolution of the emission bands in the prototypical PDR NGC 7023. We used near-IR spectra obtained with the IRC instrument onboard AKARI to trace the evolution of the 3.3 µm and 3.4 µm bands, which are associated with aromatic and aliphatic C-H bonds on PAHs. The spectral fitting involved an additional broad feature centered at 3.45 µm that is often referred to as the plateau. Mid-IR observations obtained with the IRS instrument onboard the Spitzer Space Telescope were used to distinguish the signatures of eVSGs and neutral and cationic PAHs. We correlated the spatial evolution of all these bands with the intensity of the UV field given in units of the Habing field G0 to explore how their carriers are processed. RESULTS: The intensity of the 3.45 µm plateau shows an excellent correlation with that of the 3.3 µm aromatic band (correlation coefficient R = 0.95) and a relatively poor correlation with the aliphatic 3.4 µm band (R=0.77). This indicates that the 3.45 µm feature is dominated by the emission from aromatic bonds. We show that the ratio of the 3.4 µm and 3.3 µm band intensity (I3.4/I3.3) decreases by a factor of 4 at the PDR interface from the more UV-shielded layers (G0 ~ 150, I3.4/I3.3 = 0.13) to the more exposed layers (G0 > 1 × 104, I3.4/I3.3 = 0.03). The intensity of the 3.3 µm band relative to the total neutral PAH intensity shows an overall increase with G0, associated with an increase of both the hardness of the UV field and the H abundance. In contrast, the intensity of the 3.4 µm band relative to the total neutral PAH intensity decreases with G0, showing that their carriers are actively destroyed by UV irradiation and are not efficiently regenerated. The transition region between the aliphatic and aromatic material is found to correspond spatially with the transition zone between neutral PAHs and eVSGs. CONCLUSIONS: We conclude that the photo-processing of eVSGs leads to the production of PAHs with attached aliphatic sidegroups that are revealed by the 3.4 µm emission band. Our analysis provides evidence for the presence of very small grains of mixed aromatic and aliphatic composition in PDRs.
RESUMO
Emotional stress activates oxytocin neurons in the hypothalamic supraoptic and paraventricular nuclei and stimulates oxytocin release from the posterior pituitary. Oxytocin neurons in the hypothalamus have synaptic contact with prolactin-releasing peptide (PrRP) neurons. Intracerebroventricular administration of PrRP stimulates oxytocin release from the pituitary. These observations raise the possibility that PrRP neurons play a role in oxytocin response to emotional stress. To test this hypothesis, we first examined expression of Fos protein, an immediate early gene product, in the PrRP neurons in the medulla oblongata after conditioned-fear stimuli. Conditioned-fear stimuli increased the number of PrRP cells expressing Fos protein especially in the dorsomedial medulla. In order to determine whether PrRP cells projecting to the supraoptic nucleus are activated after conditioned-fear stimuli, we injected retrograde tracers into the supraoptic nucleus. Conditioned-fear stimuli induced expression of Fos protein in retrogradely labeled PrRP cells in the dorsomedial medulla. Finally we investigated whether immunoneutralization of endogenous PrRP impairs oxytocin release after emotional stimuli. An i.c.v. injection of a mouse monoclonal anti-PrRP antibody impaired release of oxytocin but not of adrenocorticotrophic hormone or prolactin and did not significantly change freezing behavior in response to conditioned-fear stimuli. From these data, we conclude that PrRP neurons in the dorsomedial medulla that project to the hypothalamus play a facilitative role in oxytocin release after emotional stimuli in rats.
Assuntos
Condicionamento Psicológico/fisiologia , Medo/fisiologia , Hormônios Hipotalâmicos/metabolismo , Hipotálamo Anterior/metabolismo , Neurônios/efeitos dos fármacos , Neuropeptídeos/metabolismo , Ocitocina/farmacologia , Hormônio Adrenocorticotrópico/metabolismo , Animais , Anticorpos/administração & dosagem , Comportamento Animal , Contagem de Células , Corantes Fluorescentes/farmacocinética , Hemoglobinas/metabolismo , Hormônios Hipotalâmicos/imunologia , Hipotálamo Anterior/anatomia & histologia , Imuno-Histoquímica/métodos , Injeções Intraventriculares/métodos , Masculino , Bulbo/anatomia & histologia , Bulbo/metabolismo , Neurônios/metabolismo , Neuropeptídeos/imunologia , Concentração Osmolar , Hormônio Liberador de Prolactina , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ratos , Ratos Wistar , Rodaminas/farmacocinéticaRESUMO
During prolonged exposure to morphine, oxytocin neurons of the rat supraoptic nucleus develop dependence, shown by hyperexcitation following morphine withdrawal. The present study investigated the role of afferent projections to the supraoptic nucleus in this withdrawal excitation. Rats were made morphine-dependent by continuous intracerebroventricular infusion of morphine at increasing doses (up to 50 microg/h). On the sixth day, rats were anaesthetized with pentobarbitone and morphine withdrawal was precipitated by intraperitoneal injection of naloxone (5 mg/kg). Fos-immunoreactivity in the supraoptic nucleus, and also in the median preoptic nucleus, organum vasculosum of the lamina terminalis and subfornical organ, which project to the supraoptic nucleus, increased following morphine withdrawal. However, retrograde tracing from the supraoptic nucleus showed that, of the neurons in these regions which project to the supraoptic nucleus, only 0.4-7.1% expressed Fos in response to morphine withdrawal. Following morphine withdrawal, Fos-immunoreactivity was present in 39.2% and 19.8% of the tyrosine hydroxylase-immunoreactive neurons of the A1/C1 and A2/C2 cell groups. Of the cells in these regions identified as projecting to the supraoptic nucleus, 11.3% in the region of the A2 cell group and 12.7% in the region of the A1 cell group expressed Fos after morphine withdrawal. In a second study, monoamine release was measured in the supraoptic nucleus of urethane-anaesthetized morphine-dependent and -naive rats. Retrodialysis of naloxone (10[-5] M) into the supraoptic nucleus induced a small increase in plasma oxytocin concentration in morphine-dependent rats (13.5+/-4.8 pg/ml increase) but not in naive rats (1.2+/-5.9 pg/ml decrease), with no significant change in monoamine release in either morphine-dependent or -naive rats. Intravenous injection of naloxone (5 mg/kg) 1 h later produced a further significant increase in plasma oxytocin concentration in morphine-dependent rats concomitant with a significant increase in noradrenaline release from the supraoptic nucleus. Thus, morphine-withdrawal excitation of supraoptic oxytocin neurons occurs concurrently with a modestly increased activity of their input from the brainstem, and very little activation in other known inputs.
Assuntos
Morfina/farmacologia , Naloxona/farmacologia , Antagonistas de Entorpecentes/farmacologia , Entorpecentes/farmacologia , Neurônios Aferentes/fisiologia , Ocitocina/fisiologia , Síndrome de Abstinência a Substâncias/fisiopatologia , Núcleo Supraóptico/fisiologia , Vias Aferentes/citologia , Vias Aferentes/fisiologia , Animais , Tronco Encefálico/efeitos dos fármacos , Tronco Encefálico/metabolismo , Tronco Encefálico/fisiologia , Feminino , Imuno-Histoquímica , Microdiálise , Dependência de Morfina/fisiopatologia , Neurônios Aferentes/efeitos dos fármacos , Neurônios Aferentes/metabolismo , Norepinefrina/metabolismo , Norepinefrina/farmacologia , Proteínas Proto-Oncogênicas c-fos/biossíntese , Ratos , Ratos Wistar , Núcleo Supraóptico/citologia , Núcleo Supraóptico/metabolismoRESUMO
Activation of abdominal vagal afferents by peripheral injection of cholecystokinin octapeptide induces oxytocin release into the circulation. To test the hypothesis that cholecystokinin increases oxytocin release via activation of noradrenergic afferents from the brainstem, we injected rats with 5-amino-2,4-dihydroxy-alpha-methylphenylethylamine, a selective neurotoxin to noradrenergic fibres, into a lateral cerebral ventricle. The neurotoxin treatment reduced the noradrenaline content in the hypothalamus by 75% and reduced the oxytocin secretion in response to cholecystokinin by over 90%. In separate experiments, the neurotoxin was injected unilaterally in the vicinity of the supraoptic nucleus to test whether direct noradrenergic afferents to the supraoptic nucleus are involved in the response to cholecystokinin. The injection reduced the immunoreactivity for dopamine beta-hydroxylase in the supraoptic nucleus and significantly decreased the number of the supraoptic neurons expressing Fos-like protein after cholecystokinin but not after hypertonic saline. In further experiments, rhodamine-conjugated latex microspheres were injected into the supraoptic nucleus to retrogradely label afferent neurons, and the brains were processed with double-immunohistochemistry for tyrosine hydroxylase and Fos-like protein. In the C2/A2 but not the C1/A1 region of the brainstem, cholecystokinin increased the expression of Fos-like protein in the population of retrogradely-labelled catecholaminergic cells. In the C2/A2 region, the majority of retrogradely labelled cells expressing Fos-like protein after cholecystokinin were catecholaminergic. We conclude that noradrenergic afferents from the A2 but not from the A1 region of the brainstem to the hypothalamus mediate, at least in part, oxytocin release following cholecystokinin.
Assuntos
Ocitocina/metabolismo , Sincalida/farmacologia , Núcleo Solitário/citologia , Núcleo Supraóptico/citologia , Vias Aferentes/citologia , Animais , Imuno-Histoquímica , Injeções Intraventriculares , Masculino , Microesferas , Neurônios/química , Neurotoxinas/farmacologia , Norepinefrina/metabolismo , Fenetilaminas/farmacologia , Proteínas Proto-Oncogênicas c-fos/imunologia , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ratos , Ratos Wistar , Rodaminas , Núcleo Solitário/metabolismo , Núcleo Supraóptico/metabolismoRESUMO
Oxytocin is released from the pituitary gland in response to a variety of stressful stimuli, including noxious stimuli, conditioned fear and exposure to novel environments. These responses are believed to be mediated, at least in part, by noradrenergic projections from the medulla oblongata, and some of these noradrenergic neurones also contain prolactin-releasing peptide (PrRP). Central administration of either PrRP or noradrenaline stimulates oxytocin secretion into the circulation. Stressful stimuli activate PrRP-containing noradrenergic neurones in the medulla oblongata, and it is thus possible that PrRP/noradrenergic projections to the hypothalamus mediate oxytocin responses to stressful stimuli. Here, the roles of brainstem PrRP/noradrenergic projections to the hypothalamus in oxytocin responses to different kinds of stressful stimuli are reviewed, with a particular emphasis on conditioned fear. Roles of dendritic oxytocin release during stress and metabolic factors affecting stress pathways are also discussed.
Assuntos
Bulbo/citologia , Vias Neurais/fisiologia , Ocitocina/metabolismo , Hipófise/citologia , Estresse Fisiológico/metabolismo , Animais , Bulbo/metabolismo , Hipófise/metabolismoRESUMO
Emotional responsiveness is reduced in adult animals which have been exposed to stress during their first few weeks of life (1, 2). The stress of 'handling', daily removal of pups from their mother during the pre-weaning period, also leads to a reduced corticosterone response to novel stimuli in adult life (3). In rats, exposure to novel stimuli results in the concomitant release of prolactin (PRL) and corticosterone (4-6). Here we show that, in male rats handled daily during the pre-weaning period of life and tested in adult life for their hormonal responses to exposure to novel audio-visual stimuli, the consequent secretion of adrenocorticotrophin (ACTH) is attenuated, but that of PRL is not. Thus, pre-weaning handling results in permanent changes in a neural system specific to the control of ACTH secretion rather than affecting pathways common to neuroendocrine responses to emotional stimuli (7).
RESUMO
The present study aimed to examine roles of N-methyl-D-aspartic acid (NMDA) receptors in oxytocin and vasopressin release after osmotic stimuli. A noncompetitive NMDA receptor antagonist, MK-801 (0.2 mg/kg body weight, i.p.), significantly decreased plasma concentrations of oxytocin and vasopressin after hypertonic saline injection (0.3 or 0.6 M NaCl, i.p., 20 ml/kg). By contrast, oxytocin release induced by injection of cholecystokinin octapeptide (20 microg/kg, i.p.) was not significantly changed by MK-801. Hypertonic saline injection increased the number of cells expressing Fos in the supraoptic nucleus and in the regions anterior and ventral to the third ventricle (AV3V) regions [the organum vasculosum of the lamina terminalis (OVLT) and median preoptic nucleus]. MK-801 decreased the number of cells expressing protein in these areas after hypertonic saline injection. A microdialysis method showed that a hypertonic saline injection (0.6 M NaCl, 20 ml/kg, i.p.) facilitated glutamic acid release in and near the OVLT. The results support the view that NMDA receptor in the AV3V region modulates in a facilitative fashion the AV3V inputs to the supraoptic neurosecretory neurones.
Assuntos
Ocitocina/metabolismo , Receptores de N-Metil-D-Aspartato/fisiologia , Solução Salina Hipertônica/farmacologia , Vasopressinas/metabolismo , Animais , Colecistocinina/farmacologia , Maleato de Dizocilpina/farmacologia , Antagonistas de Aminoácidos Excitatórios/farmacologia , Ácido Glutâmico/metabolismo , Hipotálamo/metabolismo , Injeções Intraperitoneais , Concentração Osmolar , Ocitocina/sangue , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ratos , Vasopressinas/sangueRESUMO
Effects of novel environmental stimuli on vasopressin and oxytocin secretion by the pituitary were studied in dehydrated male rats. As the novel environmental stimuli, rats were transferred to an experimental room, placed in a box painted black and given a pure tone auditory stimulus of 2 kHz. Exposure of rats to the novel environmental stimuli for a period of 2 min decreased plasma concentrations of vasopressin and increased plasma levels of adrenocorticotrophic hormone (ACTH) and prolactin, but did not significantly change the plasma level of oxytocin. The stimuli, however, became ineffective for producing the suppressive vasopressin response as the period of exposure was prolonged to more than 5 and up to 30 min, although the prolonged stimuli were still effective for inducing facilitatory ACTH and prolactin responses. After repeated exposures of rats to the environmental stimuli once a day for 5 or 10 days, the stimuli became disabled from producing the suppressive vasopressin response. However, the rats were still capable of responding to the novel stimuli of another kind. All these data suggest that novelty stress suppresses vasopressin secretion but does not change oxytocin secretion. In order to test the possibility that glucocorticoids expectedly secreted by the adrenals in response to the stress might have suppressed vasopressin secretion, a large amount of dexamethasone was administered to the rat before testing. Dexamethasone pretreatment depressed plasma levels of ACTH and vasopressin as reported previously and blocked the facilitatory ACTH response to the novelty stress. However, dexamethasone treatment did not affect the suppressive vasopressin response to the novelty stress.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Ocitocina/metabolismo , Hipófise/metabolismo , Estresse Psicológico/fisiopatologia , Vasopressinas/metabolismo , Estimulação Acústica , Hormônio Adrenocorticotrópico/metabolismo , Animais , Dexametasona/farmacologia , Masculino , Prolactina/metabolismo , Ratos , Ratos WistarRESUMO
Intravenously administered cholecystokinin octapeptide (CCK) induces oxytocin release from the neurohypophysis in anaesthetised rats. Memory of conditioned taste aversion can be acquired under anaesthesia. The present experiments aimed at investigating whether taste stimuli previously paired with i.v. CCK evoke oxytocin release from the neurohypophysis in urethane-anaesthetised male rats. Sucrose solution (0.75-2.0 M) paired with i.v. CCK or the vehicle was applied to the tongue. After 3 h, sucrose solution was applied again. The second sucrose slightly increased plasma oxytocin concentration in rats that had received the first sucrose solution paired with the vehicle. Plasma oxytocin concentration after the second sucrose application, however, was significantly higher in CCK-injected than in vehicle-injected rats. In rats that received CCK 1 h before the first sucrose application, a second sucrose application did not produce the oxytocin response. The magnitude of the oxytocin response to the second sucrose solution was increased in a manner related to CCK doses. In separate experiments, NaCl solution (0.75 M) paired with CCK or the vehicle was applied to the tongue. The second NaCl solution applied 3 h after the first one facilitated oxytocin release both in the rats that had received CCK or the vehicle. The increase in plasma oxytocin, however, was significantly larger in CCK than in vehicle-injected rats. In rats that had received the first sucrose solution paired with CCK, a second sucrose solution evoked a significantly larger increase in plasma oxytocin concentrations than a testing NaCl solution did. In rats that had received NaCl solution paired with CCK, a testing sucrose solution did not significantly change plasma oxytocin concentrations. These data suggest that the taste stimulus previously paired with i.v. CCK induces oxytocin release from the neurohypophysis in urethane-anaesthetised rats.
Assuntos
Aprendizagem da Esquiva/fisiologia , Colecistocinina/farmacologia , Condicionamento Operante/fisiologia , Ocitocina/metabolismo , Neuro-Hipófise/efeitos dos fármacos , Paladar/fisiologia , Anestésicos Intravenosos , Animais , Injeções Intravenosas , Masculino , Concentração Osmolar , Neuro-Hipófise/metabolismo , Ratos , Taxa Secretória/efeitos dos fármacos , Taxa Secretória/fisiologia , Cloreto de Sódio/farmacologia , Estimulação Química , UretanaRESUMO
We examined the role of N-methyl-d-aspartate (NMDA) receptors in the control of noradrenaline release in the supraoptic nucleus (SON) using a microdialysis method in urethane-anaesthetized rats. Local application of 0.5 mm NMDA into the SON by retrodialysis decreased noradrenaline content in the dialysate from the SON. On the other hand, MK-801, a channel blocker of NMDA receptors, or D(-)2-amino-5-phosphonopentanoic acid (AP-5), a competitive NMDA receptor antagonist, increased the basal noradrenaline content. Tetrodotoxin did not completely block the noradrenaline increase after NMDA antagonists. Infusion of Ca2+-free solution containing Ni2+ and Cd2+, or a mixture of omega-agatoxin IVA and omega-conotoxin GVIA, voltage-sensitive Ca2+ channels blockers, did not block noradrenaline increase after AP-5, but blocked noradrenaline increase after high K+. Infusion of intracellular Ca2+ blockers, thapsigargin or TMB-8, impaired noradrenaline increase after AP-5 but not that after high K+. These data are consistent with the hypothesis that activation of an NMDA receptor inhibits an intracellular Ca2+ store-dependent noradrenaline release from nerve terminals in the SON.
Assuntos
Sinalização do Cálcio/fisiologia , Cálcio/fisiologia , Antagonistas de Aminoácidos Excitatórios/farmacologia , Norepinefrina/metabolismo , Receptores de N-Metil-D-Aspartato/antagonistas & inibidores , Núcleo Supraóptico/metabolismo , Inibidores da Captação Adrenérgica/farmacologia , Antagonistas Adrenérgicos alfa/farmacologia , Animais , Bloqueadores dos Canais de Cálcio/farmacologia , Desipramina/farmacologia , Masculino , Microdiálise , N-Metilaspartato/farmacologia , Piperidinas/farmacologia , Ratos , Ratos Wistar , Núcleo Supraóptico/efeitos dos fármacos , Tetrodotoxina/farmacologiaRESUMO
Emotional stress inhibits vasopressin release from the pituitary but may facilitate its release from the dendrites in the hypothalamus. We examined effects of intermittently applied footshock upon the amount of vasopressin heteronuclear RNA in the hypothalamus. The footshock decreased plasma vasopressin concentration but increased its extracellular concentration within the supraoptic nucleus. The contents of the vasopressin heteronuclear RNA in the supraoptic nucleus were significantly decreased after the shock. These data suggest that intermittent footshock decreases not only vasopressin release from the axon terminals in the pituitary, but also vasopressin synthesis in the cell bodies in the hypothalamus while the stimulus facilitates vasopressin release from the dendrites in the hypothalamus. The data also suggest differential control of dendritic vasopressin release and synthesis in the hypothalamus.
Assuntos
Dendritos/metabolismo , Núcleo Supraóptico/metabolismo , Vasopressinas/biossíntese , Vasopressinas/metabolismo , Animais , Eletrochoque , Extremidades , Masculino , RNA/análise , Ratos , Ratos Wistar , Núcleo Supraóptico/química , Núcleo Supraóptico/ultraestrutura , Vasopressinas/genéticaRESUMO
The effect of electrically evoked dendritic vasopressin release on noradrenaline release into the hypothalamic supraoptic nucleus was assessed by in vivo microdialysis in conjunction with high pressure liquid chromatography and electrochemical detection. Electrical activation of magnocellular supraoptic neurones by stimulation of their axons at the level of the neural lobe significantly increased noradrenaline release into the nucleus (2.5-fold, P<0.03). This increase was completely blocked by administration of a nonpeptide vasopressin V1a receptor antagonist via the microdialysis probe. These data suggest that dendritically released vasopressin facilitates noradrenaline release into the hypothalamic nucleus.