RESUMO
The central actions of oxytocin on reproduction-related functions and behaviors are strongly steroid-dependent and gender specific. This study characterizes sexual differences in the oxytocin binding site expression in forebrain and spinal cord of the rat. Using film autoradiography, we quantified the density of oxytocin binding sites in the ventromedial hypothalamic nucleus, the medial and central nuclei of the amygdala, the medial bed nucleus of the stria terminalis and the spinal cord dorsal horns both in adult male and female rats, and during development. In addition, neonatal castrated males and intact neonatal females treated with a single injection of testosterone (1 mg) were examined. Data showed a sexual dimorphism in the expression of oxytocin binding sites in the spinal cord dorsal horns and in restricted areas of the forebrain that are sensitive to gonadal steroids such as the ventromedial hypothalamic nucleus, but not in gonadal steroid insensitive sites such as the central nucleus of the amygdala. Adult males had higher oxytocin binding site densities in the ventromedial hypothalamic nucleus and dorsal horns than females. In the forebrain, but not in the dorsal horn, this sexual difference required a perinatal exposure to testosterone. Neonatal castration only abolished the sexual difference in the ventromedial hypothalamic nucleus of adults, but not in the dorsal horn. Furthermore, females that received a single injection of testosterone 1 day after birth showed significant increases in the density of oxytocin binding sites in the ventromedial hypothalamic nucleus, medial nucleus of the amygdala and medial bed nucleus of the stria terminalis. In addition, the findings suggest that the sexual difference in the ventromedial hypothalamic nucleus also requires gonadal hormones in adulthood. Our data support the hypothesis that sexually dimorphic oxytocin binding sites may contribute to the regulatory central actions of oxytocin in gender specific functions and behaviors such as nociception and reproduction.
Assuntos
Prosencéfalo/metabolismo , Receptores de Ocitocina/metabolismo , Medula Espinal/metabolismo , Animais , Animais Recém-Nascidos , Autorradiografia , Feminino , Masculino , Orquiectomia , Prosencéfalo/crescimento & desenvolvimento , Ratos , Ratos Sprague-Dawley , Caracteres Sexuais , Medula Espinal/crescimento & desenvolvimento , Testosterona/administração & dosagem , Testosterona/farmacologiaRESUMO
The localization of oxytocin (OT) binding sites and vasopressin (VP) binding sites of the V1a subtype was investigated by radioautography in kidneys of rabbits, mice and meriones during postnatal development and in the adult, and in the human kidney. Kidney sections were incubated in the presence of selective radioiodinated OT and V1a antagonists, respectively. The localizations were compared with those previously described in the rat. The main finding of the study was the almost constant presence in the cortex of V1a binding sites in the connecting tubule, the cortical collecting duct and in the juxtaglomerular apparatus (on the intra- and extraglomerular mesangium and the afferent arteriole). This distribution suggests an interaction of VP via V1a receptors and the kallikrein-kinin system in the kidney. OT binding sites, in comparison with V1a binding sites, were fewer and less constantly detectable in the kidney of the different species. In the mouse, their presence on the limbs of Henle's loop in the medulla points to the possibility of their involvement in the medullary concentrating process. In the kidneys of the various species, OT and V1a binding sites occurred always in differential structures. In contrast, in the human kidney cortex, a colocalization of OT and V1a binding sites was almost constantly observed. This raises the question as to the specificity of the neurohypophysial hormone receptors in the human kidney.
Assuntos
Rim/química , Rim/crescimento & desenvolvimento , Ocitocina/análise , Receptores de Vasopressinas/análise , Adulto , Animais , Animais Recém-Nascidos/metabolismo , Autorradiografia/métodos , Sítios de Ligação , Secções Congeladas/métodos , Gerbillinae , Humanos , Camundongos , Ocitocina/metabolismo , Coelhos , Receptores de Vasopressinas/metabolismoRESUMO
Using combined double immunofluorescence and laser confocal microscopy, we studied the common cellular localization of cholinergic muscarinic receptors (mAChRs) and aquaporin-4 water channels (AQP4) in the cortex, the corpus callosum and in ependymal cells of the rat brain. In the cortex, AQP4 staining was restricted to the perivascular end-feet of astrocytes. It was more widely distributed on the astrocytes of the corpus callosum. On astrocytes, mAChRs were often present in regions immunoreactive to AQP4. Ependymal cells bordering the third ventricle were also stained by both antibodies. The double staining of mAChRs with AQP4 on two different cell-types might indicate that further interactions exist which may be important in the regulation of water and electrolyte movements in the brain.
Assuntos
Aquaporinas/análise , Aquaporinas/fisiologia , Astrócitos/química , Epêndima/citologia , Receptores Muscarínicos/análise , Receptores Muscarínicos/fisiologia , Animais , Aquaporina 4 , Astrócitos/fisiologia , Córtex Cerebral/química , Córtex Cerebral/citologia , Corpo Caloso/química , Corpo Caloso/citologia , Epêndima/química , Masculino , Ratos , Ratos Wistar , Equilíbrio Hidroeletrolítico/fisiologiaRESUMO
In the magnocellular nuclei of the hypothalamus, there is a rich vascular network for which the function remains to be established. In the supraoptic nucleus, the high vascular density may be one element, which together with the water channel aquaporin-4 expressed in the astrocytes, is related to a role in osmoreception. We tested the osmoreception hypothesis by studying the correlation between vascular and cellular densities in the paraventricular nucleus and the supraoptic nucleus. Whether aquaporin-4 is likely to contribute to osmoreception was tested by studying the distribution in the magnocellular nuclei of the hypothalamus. The high vascular density may also reflect a high metabolic activity due to the synthesis of vasopressin and oxytocin. This metabolic hypothesis was tested by studying the regional cytochrome oxidase histochemistry, the local cerebral blood flow, and the density of glucose transporter type-1 in the supraoptic and paraventricular nuclei. All the magnocellular nuclei were characterized by an extended and intense aquaporin-4 labelling and a weak cytochrome oxidase histochemistry. The highest vascular density was found in the supraoptic nucleus and the magnocellular regions of the paraventricular nucleus. The local cerebral blood flow rates were surprisingly low in the paraventricular nucleus and the supraoptic nucleus in comparison to the cerebral cortex. Furthermore in these nuclei, the antibody for glucose transporter type-1 revealed two populations of vessels differing by their labelling intensity. The similarities observed between the different nuclei suggest that, in the hypothalamus, all magnocellular regions sense the plasma osmolarity. The low local cerebral blood flow, and the patterns of glucose transporter type-1 labelling and cytochrome oxidase histochemistry suggest that the high vascularization of these hypothalamic nuclei is not related to a high metabolic capacity in basal conditions.
Assuntos
Núcleo Hipotalâmico Paraventricular/irrigação sanguínea , Núcleo Supraóptico/irrigação sanguínea , Animais , Aquaporina 4 , Aquaporinas/metabolismo , Biomarcadores , Vasos Sanguíneos/anatomia & histologia , Vasos Sanguíneos/metabolismo , Circulação Cerebrovascular , Metabolismo Energético , Transportador de Glucose Tipo 1 , Histocitoquímica , Masculino , Proteínas de Transporte de Monossacarídeos/metabolismo , Ratos , Ratos Wistar , Distribuição TecidualRESUMO
In the rat, spinal autonomic neurons controlling penile erection receive descending pathways that modulate their activity. The paraventricular nucleus of the hypothalamus contributes oxytocinergic fibers to the dorsal horn and preganglionic sympathetic and parasympathetic cell columns. We used retrograde tracing techniques with pseudorabies virus combined with immunohistochemistry against oxytocin and radioligand binding detection of oxytocinergic receptors to evidence the oxytocinergic innervation of thoracolumbar and lumbosacral spinal neurons controlling penile erection. Spinal neurons labelled with pseudo-rabies virus transsynaptically transported from the corpus cavernosum were present in the intermediolateral cell column and the dorsal gray commissure of the thoracolumbar and lumbosacral spinal cord. Confocal laser scanning microscopic observation of the same preparations revealed close appositions between oxytocinergic varicosities and pseudorabies virus-infected neurons, suggesting strongly the presence of synaptic contacts. Electron microscopy confirmed this hypothesis. Oxytocin binding sites were present in the superficial layers of the dorsal horn, the dorsal gray commissure and the intermediolateral cell column in both the thoracolumbar and lumbosacral segments. In rats, stimulation of the paraventricular nucleus induces penile erection, but the link between the nucleus and penile innervation remains unknown. Our findings support the hypothesis that oxytocin, released by descending paraventriculo-spinal pathways, activates proerectile spinal neurons.
Assuntos
Gânglios Parassimpáticos/fisiologia , Gânglios Simpáticos/fisiologia , Ocitocina/fisiologia , Núcleo Hipotalâmico Paraventricular/fisiologia , Ereção Peniana/fisiologia , Medula Espinal/fisiologia , Animais , Autorradiografia , Gânglios Parassimpáticos/citologia , Gânglios Simpáticos/citologia , Herpesvirus Suídeo 1 , Radioisótopos do Iodo , Masculino , Microscopia Eletrônica , Neurofisinas/análise , Ocitocina/análise , Núcleo Hipotalâmico Paraventricular/química , Células do Corno Posterior/fisiologia , Células do Corno Posterior/ultraestrutura , Pseudorraiva , Ratos , Ratos Sprague-Dawley , Medula Espinal/citologiaRESUMO
The localization and pharmacological characteristics of vasopressin (VP) binding sites of the V(1a) subtype in developing and adult rat kidney were investigated by radioautography on kidney sections incubated in the presence of a radioiodinated selective V(1a) antagonist. Their localization after in vivo systemic infusion of the radioligand was also investigated. V(1a) binding sites first appear at embryonic day 16 on vascular elements. In the adult, they were localized in the cortex (vascular and tubular structures, juxtaglomerular apparatus), the outer medulla outer stripe (vasa recta) and inner stripe (thin descending limbs of short looped nephrons) and the inner medulla (collecting ducts). Data obtained in vitro were confirmed by in vivo binding at postnatal day 30 (PN30). Whatever their localizations, the V(1a) binding sites exhibited full V(1a) pharmacological profile in postnatal stages rats and in adult rats: a high affinity (nM range) for VP and for the V(1a) agonist, a lower affinity (microM range) for oxytocin and no affinity for the oxytocin agonist. The presence of V(1a) binding sites in these different structures raises the question of the putative roles of VP in modulating renal functions. A striking finding is the presence of V(1a) binding sites in the outer medullary thin descending limbs of short looped nephrons suggesting their colocalization with urea transporters.
Assuntos
Rim/crescimento & desenvolvimento , Rim/metabolismo , Receptores de Vasopressinas/metabolismo , Envelhecimento , Animais , Animais Recém-Nascidos , Autorradiografia , Sítios de Ligação , Ratos , Ratos Wistar , Distribuição TecidualRESUMO
The distribution of vasopressin and oxytocin binding sites in the central nervous system of the merione (Meriones shawi), a rodent adapted to desert life, was studied by means of conventional film radioautography at macroscopic scale and historadioautography at cellular level using radioiodinated ligands highly selective for either oxytocin or type V1 a vasopressin receptors. Both types of binding sites exhibited the same selectivity for endogenous peptides as in the rat. Distribution of oxytocin binding sites was similar in some structures (limbic system, spinal cord) to that described in the rat and in other rodents. Vasopressin binding sites were much more widely distributed in the merione than in the rat brain. In addition to locations common to most rodents (lateral septum and suprachiasmatic nucleus), in merione vasopressin binding sites occurred in several areas known to express oxytocin binding sites in the rat (olfactory system, hypothalamus). Colocalisation of vasopressin and oxytocin binding sites, which occurred in the CA1 and CA2 fields of Ammon's horns of the hippocampus, the caudate-putamen and the fundus striati of the merione, has so far not been reported in any other rodent.
Assuntos
Sistema Nervoso Central/anatomia & histologia , Sistema Nervoso Central/metabolismo , Gerbillinae/anatomia & histologia , Gerbillinae/metabolismo , Receptores de Ocitocina/metabolismo , Receptores de Vasopressinas/metabolismo , Animais , Autorradiografia , Sítios de Ligação , Encéfalo/anatomia & histologia , Química Encefálica/fisiologia , Histocitoquímica , Masculino , Ocitocina/metabolismo , Ratos , Medula Espinal/anatomia & histologia , Medula Espinal/metabolismo , Vasopressinas/metabolismoRESUMO
Anatomical studies have shown that the central amygdaloid nucleus (CeA) is reciprocally connected with the lateral bed nucleus of the stria terminalis (BSTL), both structures being major components of the central extended amygdala. The CeA also receives projections from the insular cortex (InsCx) and the paraventricular thalamic nucleus (PVT). Extracellular unit activity was recorded from neurons in the lateral CeA (CeL) in urethane anaesthetized rats and their responses were studied after electrical stimulation of the BSTL, InsCx and PVT. The spontaneous activity of CeL neurons was low (1.69 spikes/s) and 40% of recorded cells were silent. The iontophoretic application of the GABAA antagonist, bicuculline, increased the firing rate of 20% of neurons. The BSTL stimulation induced an antidromic response in 33% of the tested cells. Orthodromic responses were obtained from 83% (BSTL stimulation), 70% (InsCx stimulation) and 85% (PVT stimulation) of tested cells, some of which responded to both BSTL and InsCx or PVT stimulations. Orthodromic responses mostly consisted in 1-3 orthodromic spikes followed by an inhibition. During iontophoretic application of bicuculline, stimulation induced additional short latency orthodromic spikes, even in cells that were previously unresponsive. However, the duration of the inhibition was never reduced. These results indicate that GABAergic neurotransmission may play a dominant role in both spontaneous and evoked electrical activities in the CeL, probably mediated by local circuit cells involved in a feed-forward inhibition. This organization, along with the reciprocal connections between the CeL and the BSTL, is considered in the context of the extended amygdala.
Assuntos
Tonsila do Cerebelo/fisiologia , Mapeamento Encefálico , Córtex Cerebral/fisiologia , Núcleos Talâmicos/fisiologia , Animais , Bicuculina/farmacologia , Estimulação Elétrica , Potenciais Evocados/fisiologia , Antagonistas GABAérgicos/farmacologia , Iontoforese , Masculino , Vias Neurais/fisiologia , Ratos , Ratos Wistar , Transmissão Sináptica/fisiologia , Ácido gama-Aminobutírico/fisiologiaRESUMO
The functional characteristics of binding sites for the neuropeptide oxytocin (OT) detected by radioautography in laminae I and II of the dorsal horn (DH) and on cultured neonatal DH neurons were studied on the latter using perforated patch-clamp recordings. The neurons were identified by their spike discharge properties and on the basis of the presence of met-enkephalin-like and glutamate decarboxylase-like immunoreactivities. OT (100 nM) never induced any membrane current at a holding potential of -60 mV but increased the frequency of spontaneously occurring AMPA receptor-mediated EPSCs or the mean amplitude of electrically evoked EPSCs in a subset (35%) of neurons. The frequency of miniature EPSCs (m-EPSCs) recorded in the presence of 0.5 microM tetrodotoxin was also increased by OT (100 nM) without any change in their mean amplitude, indicating an action at a site close to the presynaptic terminal. The decay kinetics of any type of EPSC were never modified by OT. The effect of OT was reproduced by [Thr4, Gly7]-OT (100 nM), a selective OT receptor agonist, and blocked by d(CH2)5-[Tyr(Me)2,Thr4,Tyr-NH29]-ornithine vasotocin (100 nM), a specific OT receptor antagonist. Reducing the extracellular Ca2+ concentration from 2.5 to 0.3 mM in the presence of Cd2+ (100 microM) reversibly blocked the effect of OT on m-EPSCs. The OT receptors described here may represent the substrate for modulatory actions of descending hypothalamo-spinal OT-containing pathways on the nociceptive system.
Assuntos
Neurônios/química , Ocitocina/análogos & derivados , Receptores de AMPA/fisiologia , Medula Espinal/citologia , Transmissão Sináptica/efeitos dos fármacos , 6-Ciano-7-nitroquinoxalina-2,3-diona/farmacologia , Animais , Animais Recém-Nascidos , Benzodiazepinas/farmacologia , Cálcio/farmacologia , Estimulação Elétrica , Encefalina Metionina/análise , Encefalina Metionina/imunologia , Potenciais Evocados/efeitos dos fármacos , Potenciais Evocados/fisiologia , Antagonistas de Aminoácidos Excitatórios/farmacologia , Glutamato Descarboxilase/análise , Glutamato Descarboxilase/imunologia , Ácido Glutâmico/fisiologia , Imuno-Histoquímica , Neurônios/enzimologia , Nociceptores/fisiologia , Ocitocina/farmacologia , Técnicas de Patch-Clamp , Ratos , Ratos Wistar , Tetrodotoxina/farmacologiaRESUMO
The central amygdaloid nucleus and the lateral bed nucleus of the stria terminalis are two similar telencephalic structures belonging to the central extended amygdala. These regions contain numerous peptidergic and GABAergic neurones which maintain the neurones projecting to the brain stem under tight intrinsic control. Using immunocytochemistry in colchicine-treated rats, we showed that, in the lateral subdivision of the central amygdaloid nucleus and in the dorsal part of the lateral bed nucleus of the stria terminalis, a population of GABAergic neurones is able to co-synthesize either corticotropin-releasing factor or methionine-enkephalin, but never both peptides. These results suggest that, in the GABAergic intrinsic circuits of the central extended amygdala, co-liberated peptides can have a modulatory role on GABAergic actions.
Assuntos
Tonsila do Cerebelo/química , Hormônio Liberador da Corticotropina/análise , Encefalina Metionina/análise , Neurônios/química , Ácido gama-Aminobutírico/análise , Tonsila do Cerebelo/citologia , Animais , Imuno-Histoquímica , Masculino , Ratos , Ratos WistarRESUMO
Radioligand receptor autoradiography has shown that oxytocin- and vasopressin-binding sites exist in numerous rat brain regions, among which the amygdala and the bed nucleus of the stria terminalis (BST) are especially prominent. However, these descriptions did not take into account the numerous subdivisions of the amygdala and the BST. Thus, we have reinvestigated the distribution of these sites in the rat extended amygdala, which is formed by a continuum of structures stretching from the BST to the centromedial amygdala, including parts of the accumbens nucleus, substantia innominata, and transition areas between the amygdala and the striatum. For this purpose, histoautoradiography was used to detect binding sites at the cellular level, and anatomical boundaries were defined on the basis of acetylcholinesterase histochemistry and tyrosine-hydroxylase immunohistochemistry. Oxytocin- and vasopressin-binding sites were detected in well-defined subdivisions of both medial and central parts of the extended amygdala, but they almost never coexisted in the same region. Compared with previously reported distributions, our reinvestigation describes novel oxytocin- and vasopressin-binding sites in the lateral and supracapsular BST, in the sublenticular extended amygdala, in the interstitial nucleus of the posterior limb of the anterior commissure, in the marginal zone, in the central amygdaloid nucleus, and in the anterior amygdaloid area. These results indicate that oxytocin- and vasopressin-binding sites represent an important feature of the extended amygdala and may participate in the large variety of functions that characterize this area, including reproductive and ingestive behaviors, conditioned fear and autonomic regulation.
Assuntos
Tonsila do Cerebelo/química , Ocitocina/análise , Ratos Wistar/fisiologia , Vasopressinas/análise , Acetilcolinesterase/análise , Tonsila do Cerebelo/enzimologia , Animais , Autorradiografia , Sítios de Ligação/fisiologia , Imuno-Histoquímica , Masculino , Núcleo Accumbens/química , Núcleo Accumbens/enzimologia , Condutos Olfatórios/química , Condutos Olfatórios/enzimologia , Ocitocina/metabolismo , Ratos , Tirosina 3-Mono-Oxigenase/análise , Vasopressinas/metabolismoRESUMO
The localization of oxytocin (OT)-binding sites in the developing rat kidney and their pharmacological characterization were investigated by means of autoradiographic techniques. The cellular localization was studied by application of the histoautoradiographic technique to (1) frozen sections and semithin sections from kidney slices incubated in vitro in the presence of a 125I-labelled OT antagonist and (2) frozen and semithin sections from kidneys after in vivo systemic infusion of the radioligand. Pharmacological characteristics were determined in competition experiments by using quantitative film autoradiography. Specific OT-binding sites were first detected at embryonic day 17 (E17) in the cortex. At early stages up to postnatal days (PN30), the cortical OT-binding sites were highly concentrated on the juxta- and paraglomerular portion of the distal tubule; in the adult they were restricted to the macula densa. In the medulla, OT-binding sites were first detected at E19 when this region is forming; they were localized on the thin limb of Henle's loop. These data obtained by in vivo binding were confirmed by in vivo binding at PN30 which showed, in addition, the presence in one rat of OT-binding sites in the inner stripe of the outer medulla. At all stages examined (PN15 to PN90), cortical OT-binding sites had a higher selectivity for OT versus vasopressin (IC50 = 0.78 +/- 0.04 nM and 8 +/- 0.5 nM respectively at PN90) than medullary sites (IC50 = 1.9 +/- 0.27 nM and 2 +/- 1.13 nM respectively at PN90). These data suggest that the OT-binding sites of the macula densa and thin Henle's loop, detected in the rat kidney, represent two subtypes of OT receptors which could mediate distinct effects of OT on kidney function.
Assuntos
Rim/metabolismo , Ocitocina/metabolismo , Receptores de Ocitocina/metabolismo , Animais , Autorradiografia , Ligação Competitiva , Idade Gestacional , Técnicas In Vitro , Rim/efeitos dos fármacos , Rim/embriologia , Masculino , Ensaio Radioligante , Ratos , Ratos Wistar , Receptores de Ocitocina/análise , Vasotocina/análogos & derivados , Vasotocina/farmacologiaRESUMO
Vasopressin (VP), given intracerebrally to rats, can induce antipyresis, motor disturbances, and increases in arterial blood pressure. The possibility that the VP-binding sites in the fundus striati (FStr) could participate in these effects was investigated. After a bilateral injection of 100 pmol of VP into the FStr, the fever induced by an injection of the lipopolysaccharide of Escherichia coli (50 micrograms/kg ip) was not affected. Bilateral injections of 100 pmol of VP did not induce motor disturbances or alterations in body temperature after either of two successive injections. In contrast, bilateral injections of VP into urethan-anesthetized rats induced dose-dependent increases in arterial pressure without affecting heart rate. This increase was blocked by a V1 antagonist; oxytocin and a V2 agonist were ineffective. In keeping with this preliminary pharmacological analysis, radio-ligand-binding studies of the FStr revealed binding sites in the FStr exhibiting a binding profile typical of the V1 subtype. This study suggests that the V1 receptors in the FStr could participate in the central regulation of the blood pressure.
Assuntos
Pressão Sanguínea/fisiologia , Corpo Estriado/fisiologia , Receptores de Vasopressinas/fisiologia , Animais , Autorradiografia , Sítios de Ligação , Pressão Sanguínea/efeitos dos fármacos , Regulação da Temperatura Corporal/efeitos dos fármacos , Regulação da Temperatura Corporal/fisiologia , Febre/fisiopatologia , Frequência Cardíaca/efeitos dos fármacos , Masculino , Atividade Motora/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Ratos Wistar , Vasopressinas/metabolismo , Vasopressinas/farmacologiaRESUMO
The localization at the cellular level and the regulation by progesterone of the estrogen-sensitive oxytocin binding sites was studied in the rat telencephalon and the hypothalamus by using quantitative film-autoradiography and histoautoradiography. Male rats (castrated or not) and ovariectomized females (estradiol supplemented or not) were used to characterize these sites and to precise their localization. They were detected in the striatal cell bridges, the olfactory tubercle, the principal nucleus of the bed nucleus of the stria terminalis and the medial nucleus of the amygdala of the telencephalon and in the medial preoptic, the ventromedial and the ventral premammillary nuclei of the hypothalamus. Estrogen administration in addition induced expression of oxytocin binding sites in the major island of Calleja, the anterior hypothalamic area and the terete nucleus. The density of the estrogen-sensitive oxytocin binding sites varied during the estrous cycle, but differently in the telencephalon and the hypothalamus. In the telencephalon it peaked at proestrus 9 h and was already decreased at proestrus 21 h, whereas in the hypothalamus it was similarly high at proestrus 9 h and proestrus 21 h, suggesting the intervention of progesterone in the regulation of the hypothalamic estrogen-sensitive oxytocin binding sites.
Assuntos
Estradiol/farmacologia , Hipotálamo/efeitos dos fármacos , Progesterona/fisiologia , Receptores de Ocitocina/efeitos dos fármacos , Telencéfalo/efeitos dos fármacos , Sequência de Aminoácidos , Animais , Autorradiografia/métodos , Feminino , Masculino , Dados de Sequência Molecular , Ratos , Ratos WistarRESUMO
Oxytocin (OT) is known to exert a facilitatory effect on its own pulsatile release during the milk ejection reflex. Several experimental approaches strongly suggested that this effect occurred in the supraoptic and paraventricular magnocellular hypothalamic nuclei where the OT neurones are concentrated. In these nuclei autoradiographic techniques failed however to detect OT receptors. In the presently reported data, OT binding sites were visualized in the supraoptic, paraventricular and accessory magnocellular nuclei in lactating rats, suckled or not, after acute intracerebroventricular injection of an OT antagonist. No such labelling occurred either in untreated lactating rats or in OT antagonist treated male rats. The combined application of histoautoradiographic and immunocytochemical techniques revealed that the OT binding sites were concentrated on magnocellular OT cell bodies and dendrites but not on their axons and endings in the pituitary neural lobe. The presently detected somatodendritic autoreceptors on OT neurones probably mediate the facilitatory effect of OT on its own release during the milk ejection reflex.
Assuntos
Autorreceptores/fisiologia , Ocitocina/fisiologia , Animais , Sítios de Ligação , Feminino , Masculino , Ejeção Láctea/fisiologia , Neurônios/fisiologia , Núcleo Hipotalâmico Paraventricular/fisiologia , Gravidez , Ratos , Reflexo/fisiologiaRESUMO
Localization of oxytocin and vasopressin binding sites has so far been studied in the rat brain by means of film autoradiographs. The availability of selective iodinated ligands with high specific activity allowed us to develop the histoautoradiographical technique and to reinvestigate at the microscopic scale the distribution of these sites in the hypothalamus. Most oxytocin binding sites were localized in delimited nuclei, e.g., the medial preoptic, the ventromedial, the ventral premammillary, the supramammillary, and the medial mammillary nuclei. In addition, a weak diffuse specific labeling occurred in the medial preoptic and the anterior hypothalamic areas. The vasopressin binding sites (of the V1a type) were detected in delimited nuclei, e.g., the suprachiasmatic, the stigmoid, and the arcuate nuclei, but they were also diffusely distributed in the lateral hypothalamic and the dorsochiasmatic areas. The locations of neurohypophysial peptides binding sites detected by light microscopy are compared with those previously obtained by film autoradiography.
Assuntos
Hipotálamo/metabolismo , Ocitocina/metabolismo , Vasopressinas/metabolismo , Animais , Autorradiografia , Sítios de Ligação , Masculino , Ratos , Ratos Wistar , Distribuição TecidualRESUMO
1. The purpose of the present study was the detection at the cellular scale of the oxytocin (OT) receptors involved in the facilitatory effect of this neuropeptide on its own release during the milk ejection reflex. 2. OT binding sites were demonstrated in brain sections by using a highly selective 125I-labelled OT antagonist detected by film- and histoautoradiography. 3. Film autoradiographs revealed the presence of OT binding sites in the hypothalamic magnocellular (supraoptic, paraventricular and anterior commissural) nuclei in lactating rats, suckled or not. This detection was only possible after acute i.c.v. injection of OT antagonist which probably induced an upregulation of the OT binding sites to autoradiographically detectable levels. 4. Combined application of histoautoradiographic and immunohistochemical techniques showed that the OT binding sites were concentrated on OT magnocellular neurones. Labelling concerned cell bodies and dendrites but not the axons and endings in the pituitary neural lobe. 5. The presently detected somatodendritic autoreceptors on OT neurones probably mediate the facilitatory effect of OT on its own release during the milk ejection reflex.
Assuntos
Lactação/fisiologia , Neurônios/metabolismo , Ocitocina/fisiologia , Receptores de Ocitocina/metabolismo , Sequência de Aminoácidos , Animais , Autorradiografia , Feminino , Hipotálamo/citologia , Hipotálamo/metabolismo , Imuno-Histoquímica , Injeções Intraventriculares , Dados de Sequência Molecular , Neurônios/ultraestrutura , Ocitocina/metabolismo , Ocitocina/farmacologia , Núcleo Hipotalâmico Paraventricular/citologia , Núcleo Hipotalâmico Paraventricular/metabolismo , Ratos , Ratos Wistar , Receptores de Ocitocina/antagonistas & inibidores , Núcleo Supraóptico/citologia , Núcleo Supraóptico/metabolismo , Vasopressinas/antagonistas & inibidoresRESUMO
Central nucleus (Ce), basomedial and medial nuclei of the amygdala (AMG), and some parts of the striato-pallidal system, present high densities of oxytocin (OT)-binding sites. In order to examine whether these OT-binding sites are functional receptors, the OT neuronal sensitivity and the presence of OT-binding sites were investigated using electrophysiological and autoradiographical techniques. To identify the AMG cells, electrical stimulation of the oval subnucleus of the bed nucleus of the stria terminalis (Ov) and of the parabrachial nucleus (Pb) were performed. Somatic and auditory sensory stimulations were also tested. OT was applied by iontophoresis during extracellular single unit recordings of cells which were localized in frontal brain sections subsequently used for histoautoradiographic detection of OT-binding sites. Cells responding to Ov nucleus stimulation were located in the AMG, mainly in the Ce nucleus, whereas those responding to Pb nucleus stimulation were distributed in the Ce nucleus and in the postero lateral part of the caudate putamen. Iontophoretic OT application excited 45% of the recorded cells (43/96) among which OT alone activated spontaneous firing rate of 30 and potentiated the L-Glutamate (GLU)-induced activation on 13. These OT-sensitive neurons were located mainly in the AMG and caudate putamen areas containing OT-binding sites. These results strongly suggest that OT-binding sites found in the AMG are functional receptors upon which OT could act as a neurotransmitter and as a neuromodulator to regulate autonomic functions.