RESUMO
Magnocellular neuroendocrine cells of the supraoptic nucleus (SON) release vasopressin (VP) systemically and locally during osmotic challenge. Although both central VP and nitric oxide (NO) release appear to reduce osmotically stimulated systemic VP release, it is unknown whether they interact locally in the SON to enhance somatodendritic release of VP, a phenomenon believed to regulate systemic VP release. In this study, we examined the contribution of VP receptor subtypes and NO to local VP release from the rat SON elicited by systemic injection of 3.5 m saline. Treatment of SON punches with VP receptor antagonists decreased osmotically stimulated intranuclear VP release. Similarly, blockade of NO production, or addition of NO scavengers, reduced stimulated VP, glutamate, and aspartate release, suggesting that local NO production and activity are critical for osmotically induced intranuclear VP and excitatory amino acid release. An increase in endogenous NO release from SON punches in response to hyperosmolality was confirmed by enzymatic NO assay. Consistent with enhanced glutamate and VP release from stimulated rat SON punches, the ionotropic glutamate receptor blocker kynurenate decreased stimulated local VP release without affecting NO release. These data suggest that NO enhances local VP release in part by facilitating local release of glutamate/aspartate and that glutamate receptor activity is required for the stimulation of local VP release by osmotic challenge. Collectively, these results suggest that local VP receptors, NO, and glutamatergic signaling mediate the amplification of intranuclear VP release during hyperosmolality and may contribute to efficient, but not exhaustive, systemic release of VP during osmoregulatory challenge.
Assuntos
Autorreceptores/metabolismo , Ácido Glutâmico/metabolismo , Sistemas Neurossecretores/metabolismo , Óxido Nítrico/metabolismo , Receptores de Vasopressinas/metabolismo , Transdução de Sinais/fisiologia , Cloreto de Sódio/farmacologia , Aminoácidos/metabolismo , Animais , Aminoácidos Excitatórios/metabolismo , Técnicas In Vitro , Sistemas Neurossecretores/citologia , Sistemas Neurossecretores/efeitos dos fármacos , Óxido Nítrico/biossíntese , Osmose , Isoformas de Proteínas/metabolismo , Ratos , Ratos Sprague-Dawley , Núcleo Supraóptico/citologia , Núcleo Supraóptico/efeitos dos fármacos , Núcleo Supraóptico/metabolismo , Vasopressinas/metabolismo , Equilíbrio Hidroeletrolítico/fisiologiaRESUMO
Central release of vasopressin (VP) by the magnocellular neuroendocrine cells (MNCs) responsible for systemic VP release is believed to be important in modulating the activity of these neurons during dehydration. Central VP release from MNC somata and dendrites is stimulated by both dehydration and pituitary adenylate cyclase activating polypeptide (PACAP). Although PACAP is expressed in MNCs, its potential role in the magnocellular response to dehydration is unexplored. The current study demonstrates that prolonged dehydration increases immunoreactivity for PACAP-27, PACAP-38, and the type I PACAP receptor in the supraoptic nucleus (SON) of the rat. In addition, PACAP stimulates local VP release in the euhydrated rat SON in vitro, and this effect is reduced by the PACAP receptor antagonist PAC(6-27) (100 nm), suggesting the participation of PACAP receptors. Concomitant with its effects on local VP release, PACAP also reduces basal glutamate and aspartate release in the euhydrated rat SON. Furthermore, somatodendritic VP release elicited by acute dehydration is blocked by PAC(6-27), suggesting that endogenous PACAP participates in this response. Consistent with this, RIA revealed that local PACAP-38 release within the SON is significantly elevated during acute dehydration. These results suggest that prolonged activation of hypothalamic MNCs is accompanied by up-regulation of PACAP and the type I PACAP receptor in these cells and that somatodendritic VP release in response to acute dehydration is mediated by activation of PACAP receptors by endogenous PACAP released within the SON. A potential role for PACAP in promoting efficient, but not exhaustive, systemic release of VP from MNCs during physiological challenge is discussed.
Assuntos
Desidratação/metabolismo , Sistemas Neurossecretores/metabolismo , Polipeptídeo Hipofisário Ativador de Adenilato Ciclase/metabolismo , Receptores de Polipeptídeo Hipofisário Ativador de Adenilato Ciclase/metabolismo , Núcleo Supraóptico/metabolismo , Vasopressinas/metabolismo , Adaptação Fisiológica , Animais , Imuno-Histoquímica , Masculino , Sistemas Neurossecretores/citologia , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/fisiologia , Núcleo Supraóptico/citologia , Equilíbrio Hidroeletrolítico/fisiologiaRESUMO
The parabrachial nucleus (PBN) is a regulatory nucleus that relays visceral information from the brain stem to the cortex. Immunohistochemical studies have shown that the levels of various neuropeptides in the PBN were changed after visceral afferent activation. Because the major transmitter relaying visceral information through the PBN is glutamate, the present study asked if glutamate release into the PBN also was changed after vagal afferent activation in anesthetized male rats. The distally crushed vagus was stimulated (50 Hz, 1-2 mA, 1 s on, 2 s off) for 2 h. Dialysates of the PBN were collected every 30 min and assayed for glutamate using high-performance liquid chromatography. Extracellular glutamate concentrations were reduced during the vagal stimulation, increased fourfold compared with prestimulated levels after the stimulation was terminated, and returned to prestimulated levels over the next 2 h poststimulation. These changes were not due to alterations in blood pressure because sodium nitroprusside infusion for the same interval resulted in a similar hypotension, but increased glutamate release. Phenylephrine and sodium nitroprusside were infused intravenously to measure the cardiac baroreflex before, during, and after vagal stimulation. The pressor (but not the depressor) response was elevated during the period of enhanced glutamate release, and the baroreflex curve was shifted to the right (increased threshold) without a change in gain. These changes in the cardiac baroreflex were reduced, but still seen, when the PBNs were dialyzed bilaterally with the glutamate antagonists MK-801 and 6-cyano-7-nitroquinoxaline-2,3-dione. Thus visceral activation alters glutamate release in the PBN and has enduring effects on cardiac baroreflex function.
Assuntos
Barorreflexo , Glutamatos/metabolismo , Ponte/fisiologia , Nervo Vago/fisiologia , Vias Aferentes , Animais , Pressão Sanguínea , Estimulação Elétrica , Frequência Cardíaca , Hipertensão/fisiopatologia , Masculino , Nitroprussiato/farmacologia , Ratos , Ratos Sprague-DawleyRESUMO
A close anatomical relationship between nerves containing substance P and calcitonin gene-related peptide (CGRP) and mast cells containing serotonin has been demonstrated in the rat lacrimal gland. This study investigates the potential for peptidergic regulation of lacrimal mast cells by examining the actions of substance P, CGRP and serotonin on protein and peroxidase secretion from isolated lacrimal segments and on substance P and CGRP to release serotonin from the lacrimal mast cells. Substance P, CGRP and serotonin evoked marked increases in total protein and peroxidase from the lacrimal. Sodium cromoglycate, a mast cell stabilizer, significantly reduced or blocked the secretory responses elicited by these agonists. Chromatographic analysis using electrochemical detection revealed that substance P, but not CGRP, augmented the release of serotonin from the gland. The substance P evoked peroxidase secretion and serotonin release was blocked by CGRP and by sodium cromoglycate. These results support a role for mast cells in the regulation of lacrimal secretion and suggest a novel regulatory interaction between substance P and CGRP in the control of lacrimal function through a neuro-immune interaction.
Assuntos
Peptídeo Relacionado com Gene de Calcitonina/farmacologia , Aparelho Lacrimal/metabolismo , Serotonina/metabolismo , Substância P/farmacologia , Animais , Cromolina Sódica/farmacologia , Aparelho Lacrimal/efeitos dos fármacos , Masculino , Mastócitos/efeitos dos fármacos , Mastócitos/enzimologia , Peroxidases/metabolismo , Ratos , Ratos Sprague-Dawley , Estimulação QuímicaRESUMO
1. Dopaminergic transmission was investigated in the central nervous system (CNS) of the freshwater snail, Lymnaea stagnalis. 2. The giant pedal neuron, designated as right pedal dorsal one (RPeD1), makes chemical, monosynaptic connections with a number of identified follower cells in the CNS. Previous work has shown that RPeD1 is an interneuron and a important component of the Lymnaea respiratory central pattern generator. In this study, the hypothesis that RPeD1 uses dopamine as its neurotransmitter was tested by chromatographic, pharmacological, and electrophysiological methods. Characterization of RPeD1's transmitter pharmacology is essential to clearly understand its role in Lymnaea. 3. Earlier studies demonstrated that the soma of RPeD1 contains dopamine. This was quantitated in the present study by high-performance liquid chromatography (with electrochemical detection) of isolated RPeD1 somata and growth cones, which yielded 0.8 +/- 0.3 and 0.10 +/- 0.08 pmol of dopamine per soma and growth cone, respectively. 4. Bath or pressure application of dopamine to follower cells of RPeD1, in situ, mimicked the effects of RPeD1 stimulation. Dose-response curves were constructed for the excitatory effect of dopamine on follower cells, visceral dorsal two and three (VD2/3) (ED50 = 39 microM; Hill coefficient = 1.03), and the inhibitory effect of dopamine on follower cell, visceral dorsal four (ED50 = 33 microM; Hill coefficient = 0.92). 5. The following dopamine agonists (100 microM) were tested by bath application: 2-amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene (ADTN), apopmorphine, 2-bromo-alpha-ergocryptine, deoxyepinephrine (DE), mesulergine, (-) quinpirole, SKF 38393, and tyramine. Only the general dopamine agonists, ADTN and DE, mimicked RPeD1's effects on its follower cells. 6. When VD2/3 was isolated and plated in vitro, it maintained a depolarizing response to dopamine. This response was reduced by intracellular injection of the G-protein blocker, GDP-beta-S (2 mM in electrode). Similarly, incubation of VD2/3, in vitro for approximately 18 h, with pertussis toxin (PTX; 5 micrograms/ml), the G-protein inactivating exotoxin, also reduced the dopamine response. Injecting GDP or incubating in heat-inactivated PTX did not effect the response. 7. Several dopamine antagonists were used in an attempt to block RPeD1's synapses: chlorpromazine, ergonovine, fluphenazine, haloperidol, 6-hydroxydopamine, SCH 23390, (+/-) sulpiride, and tubocurarine. Only the D-2 dopamine receptor antagonist, (+/-) sulpiride, reversibly blocked synaptic transmission from RPeD1 to its follower cells. Both the (+) and the (-) enantiomer of sulpiride also antagonized synaptic transmission. A dose-inhibition curve for (+/-) sulpiride was constructed (IC50 = 47 microM).(ABSTRACT TRUNCATED AT 400 WORDS)
Assuntos
Sistema Nervoso Central/fisiologia , Dopamina/metabolismo , Dopamina/farmacologia , Neurônios/fisiologia , Transmissão Sináptica/fisiologia , Animais , Células Cultivadas/efeitos dos fármacos , Sistema Nervoso Central/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Relação Dose-Resposta a Droga , Eletrofisiologia , LymnaeaRESUMO
Lactation is associated with complex changes of the hypothalamo-neurohypophysial system, and oxytocin released within the hypothalamic supraoptic (SON) and paraventricular nuclei may serve as a signal of communication between the magnocellular nuclei in lactating rats. In the first study, the intranuclear and peripheral release patterns of oxytocin and vasopressin in response to intraperitoneal hypertonic saline were studied in virgin and lactating rats to determine if the reduced osmoresponsiveness of the oxytocinergic and vasopressinergic systems during lactation is reflected by reduced release not only into blood, but also within the SON. Simultaneous microdialysis was performed within the SON and the jugular vein before and up to 6 hr after peripheral osmotic stimulation (3.0 M NaCl, 0.6 ml/100 gm body weight, i.p.). There was an immediate increase in secretion of both oxytocin and vasopressin into blood, whereas peptide release within the SON was delayed and peaked after 4-5 hr. Peripheral release of both peptides was significantly reduced in lactating animals, whereas within the SON release of oxytocin, but not vasopressin, was significantly reduced during lactation. In the second study, cross talk between the SONs--another phenomenon which seems to be characteristic for lactation--was studied. Microdialysis of one SON with hypertonic perfusion medium (with 1 M NaCl) significantly increased the release of oxytocin, vasopressin, and various amino acids (aspartate, glutamate, serine, glutamine, gamma amino butyric acid, and arginine) within the ipsilateral SON. In contrast to virgin female and male animals, this unilateral stimulation of the SON resulted in a transiently increased release of oxytocin in the contralateral SON of lactating rats. The release of vasopressin and amino acids within the contralateral SON of lactating rats remained unchanged, indicating specific activation of contralateral oxytocinergic neurons.
Assuntos
Aminoácidos/metabolismo , Arginina Vasopressina/metabolismo , Lactação/fisiologia , Ocitocina/metabolismo , Solução Salina Hipertônica/farmacologia , Núcleo Supraóptico/fisiologia , Aminoácidos/análise , Análise de Variância , Animais , Arginina Vasopressina/análise , Feminino , Lateralidade Funcional , Masculino , Microdiálise , Ocitocina/análise , Ratos , Ratos Sprague-Dawley , Caracteres SexuaisRESUMO
Recent studies have shown that the neuropeptides arginine-8-vasopressin (AVP) and oxytocin (OXT) are released within the supraoptic (SON) and paraventricular (PVN) nuclei of the hypothalamus in response to microdialysis of these nuclei with high-NaCl perfusion media. These results suggest an inherent osmosensitivity of SON and PVN neurons. To investigate whether the observed release of AVP/OXT is a unique phenomenon to these neuropeptides, several brain regions were examined for the release of amino acids or dopamine in response to high- or low-NaCl stimulation. Urethane-anesthetized male Sprague-Dawley rats were perfused with five-ion solution using U-shaped microdialysis probes. Samples were collected at 30-min intervals and analyzed for amino acids and dopamine by HPLC. In the dialysates of all perfusion areas, including the SON, PVN, hippocampus, and striatum, concentrations of Asp, Glu, Ser, Gln, Gly, taurine (Tau), and gamma-aminobutyric acid (GABA) were significantly increased during perfusion with high-NaCl medium. This release was found to be dose dependent when tested in the hippocampus and striatum with perfusion medium containing 0.5 or 1.0 M NaCl. However, only the release of Glu and Ser was found to be Ca2+ dependent. In contrast, the use of mannitol, a nonionic osmolyte, for perfusions in the striatum in concentrations of 0.5 and 1 M resulted in reduced levels of amino acids in the dialysates (Glu, Ser, Gln, and Tau). Low-NaCl perfusion medium (0.01 M) resulted in significantly increased Glu, Tau, Gly, and GABA levels in the striatum. In addition, dopamine levels in striatal dialysates were significantly increased during stimulation with 1 M NaCl. These results indicate that stimulation with high NaCl concentrations affects the release of several neurotransmitters and is not specific for AVP and OXT. The described phenomenon of the release of amino acids in response to this stimulation seems to be a response to the changed ionic concentration rather than to the osmolality. In light of these findings shown for amino acids and dopamine as well as those previously reported for AVP, OXT, and angiotensin, it would appear that sensitivity to tonicity changes brought about by microdialysis may be a feature of many transmitter systems.
Assuntos
Aminoácidos/metabolismo , Encéfalo/metabolismo , Dopamina/metabolismo , Cloreto de Sódio/farmacologia , Animais , Corpo Estriado/metabolismo , Hipocampo/metabolismo , Hipotálamo/metabolismo , Masculino , Microdiálise , Concentração Osmolar , Ratos , Ratos Sprague-Dawley , Distribuição TecidualRESUMO
We have examined potential functions of nitric oxide (NO) within the paraventricular nucleus (PVN) in urethan-anesthetized male Sprague-Dawley rats. Initial experiments demonstrated microinjection of 50 pmol of the NO donor, sodium nitroprusside (SNP), directly into the PVN resulted in significant decreases in mean blood pressure (BP) (-3,312 +/- 1,189 mmHg/s over 300-s response time; P < 0.05). To determine whether such effects were attributable to SNP-induced NO release, NO was administered into PVN directly by bilateral microdialysis of NO-containing artificial cerebrospinal fluid (NO-aCSF), a process that results in delivery of approximately 50 pmol NO.PVN-1 x min-1. Such microdialysis resulted in significant decreases in BP (-5,121 +/- 817 mmHg/s over 1,200-s response time; P < 0.005), while aCSF microdialysis was without effect (1,298 +/- 1,071 mmHg/s over 1,200-s response time; P > 0.1). Amino acid concentrations were measured in dialysates collected during perfusion of the same PVN sites with either aCSF or NO-aCSF by high-performance liquid chromatography (HPLC) analysis. NO-aCSF induced significant increases in aspartate (aCSF 31 +/- 7 pmol/30 min; NO-aCSF 134 +/- 33 pmol/30 min; P < 0.05), glutamate (aCSF 36 +/- 5 pmol/30 min; NO-aCSF 417 +/- 108 pmol/30 min; P < 0.02), gamma-aminobutyric acid (aCSF 4.1 +/- 0.7 pmol/30 min; NO-aCSF 104 +/- 29 pmol/30 min; P < 0.02), and taurine (aCSF 34 +/- 3 pmol/30 min; NO-aCSF 117 +/- 24 pmol/30 min; P < 0.01) concentrations, while alanine, glutamine, and serine concentrations were unaffected.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Aminoácidos/metabolismo , Sistema Cardiovascular/efeitos dos fármacos , Óxido Nítrico/farmacologia , Núcleo Hipotalâmico Paraventricular/efeitos dos fármacos , Núcleo Hipotalâmico Paraventricular/metabolismo , Animais , Masculino , Microdiálise , Microinjeções , Óxido Nítrico/administração & dosagem , Nitroprussiato/administração & dosagem , Nitroprussiato/farmacologia , Ratos , Ratos Sprague-Dawley , SoluçõesRESUMO
A technique, consisting of a pre-calibrated, catheter-peristaltic pump combination, for continuous blood sampling was tested using six volunteers during a 20 min immersion in cold water at 14.7 +/- 0.9 degrees C. The device offered the advantage of continued collection of blood samples from an antecubital vein during the experiment with little discomfort for the subjects and in sufficient volume for assay of plasma constituents eg. catecholamines.
Assuntos
Coleta de Amostras Sanguíneas/métodos , Imersão , Coleta de Amostras Sanguíneas/instrumentação , Temperatura Baixa , Feminino , Humanos , MasculinoRESUMO
1. We describe three interneurones and their follower cells in the central ganglionic ring of Helisoma trivolvis. 2. The largest neurone on the dorsal surface of the left pedal ganglion is shown to be an interneurone that contains dopamine and makes monosynaptic connections with a large number of follower cells in the visceral and left parietal ganglia. This neurone is designated as left pedal dorsal 1 (LPeD1). 3. Another giant neurone is located on the dorsal surface of the right pedal ganglion. Although the position and morphology of this cell, designated right pedal dorsal 1 (RPeD1), are similar to those of LPeD1, it contains serotonin rather than dopamine. This neurone was found to synapse only on LPeD1, no other follower cells have so far been discovered. The connections between LPeD1 and RPeD1 are mutually inhibitory. 4. A small FMRFamide-immunoreactive neurone, identified here as visceral dorsal 4 (VD4), is located on the dorsal surface of the visceral ganglion. This neurone has a large number of follower cells throughout the central ganglionic ring. Among these follower cells are LPeD1 and RPeD1. The transmitter utilized by VD4 at these synapses is probably FMRFamide. In addition, VD4 receives excitatory inputs from LPeD1 that appear to be chemical and monosynaptic. 5. To test further the monosynaptic and specific nature of the connections within the network, the three interneurones were isolated and cultured in vitro. In these circumstances, the three neurones extended neurites and formed synapses which, with one exception (occasional electrical coupling between LPeD1 and RPeD1), were of similar type to those observed in vivo. 6. The identification and characterization of these three interneurones and their follower cells should greatly facilitate future studies of the Helisoma trivolvis nervous system. Furthermore, the possibility that this three-cell network can be reconstructed in vitro should aid our understanding of the mechanisms underlying synapse formation and neuronal plasticity.
Assuntos
Interneurônios/citologia , Rede Nervosa/citologia , Caramujos/anatomia & histologia , Animais , Células Cultivadas , Dopamina/metabolismo , Gânglios/citologia , Interneurônios/metabolismo , Serotonina/metabolismo , Especificidade da Espécie , SinapsesRESUMO
Previous studies have shown that bilateral decentralization of the superior cervical ganglia (SCG; decentralization) attenuates allergen-induced pulmonary inflammatory responses in male rats sensitized to the nematode Nippostrongylus brasiliensis. The present report examines the neuronal and glandular mechanisms mediating the protection against pulmonary inflammation afforded by decentralization. Tissues and organs innervated by the SCG are responsible for this protection since, in a manner similar to decentralization, bilateral removal of the SCG (ganglionectomy) reduced anaphylaxis-induced accumulation of inflammatory cells in bronchoalveolar lavage fluid. Removal of the submandibular gland (sialadenectomy) did not modify the severity of the pulmonary inflammation, but concurrent sialadenectomy and decentralization abolished the protective effect of decentralization. Thus, we postulate that cervical sympathetic nerves tonically inhibit release of anti-inflammatory factors from submandibular glands. No relationship was found between noradrenaline and serotonin content of submandibular glands and the degree of protection against pulmonary inflammation offered by decentralization and ganglionectomy. Both decentralization and ganglionectomy appeared to increase the level of transcripts that encode immunomodulatory growth factors (nerve growth factor and epidermal growth factor) in submandibular glands, but these denervations evidently did not modify the transcripts for TGF beta 2. Systemic inflammatory events are regulated by the central nervous system at a level superior to the SCG probably through modulation of immunoregulatory factors in submandibular glands.
Assuntos
Anafilaxia/fisiopatologia , Pulmão/patologia , Glândula Submandibular/fisiopatologia , Anafilaxia/patologia , Animais , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/patologia , Catecolaminas/análise , Fator de Crescimento Epidérmico/biossíntese , Gânglios Simpáticos/fisiopatologia , Gânglios Simpáticos/cirurgia , Imunização , Inflamação , Masculino , Fatores de Crescimento Neural/biossíntese , Nippostrongylus/imunologia , Reação em Cadeia da Polimerase , Ratos , Ratos Endogâmicos , Glândula Submandibular/cirurgia , Simpatectomia , Fator de Crescimento Transformador beta/biossínteseRESUMO
In the present study several techniques were employed to test the hypothesis that gamma-aminobutyric acid (GABA) is a neurotransmitter in the central nervous system (CNS) of the pond snail Helisoma trivolvis (Mollusca, Pulmonata). First, by using chromatographic techniques, the presence of GABA and its differential distribution among the ganglia constituting the CNS was demonstrated. Second, de novo synthesis of 3H-GABA from 3H-glutamate was shown by the CNS. Levels of both endogenous and newly synthesized GABA were greatest in the buccal, cerebral, and pedal ganglia. Third, indirect immunohistochemistry of wholemounts revealed a central network of GABA-like immunoreactive neurons. With the possible exceptions of two pairs of fibers in nerve trunks, all projections from GABA-immunoreactive neurons were confined to the CNS, suggesting a predominantly central role for GABA. Stained neurons were found on the dorsal surface of the buccal ganglia and throughout the cerebral and pedal ganglia. No GABA-immunoreactive cell bodies were observed in the parietal, pleural, or visceral ganglia. Finally, uptake of 3H-GABA was examined autoradiographically in sectioned ganglia. A pattern of radiolabelled cells was observed that closely resembled the distribution of GABA-immunoreactive neurons. The data described above fulfill several criteria necessary to establish GABA as a transmitter in the nervous system of Helisoma. Taken together with previously obtained pharmacological evidence demonstrating that GABA acts on Helisoma central neurons, GABA is considered to be a strong candidate for a neurotransmitter in Helisoma.
Assuntos
Sistema Nervoso/metabolismo , Caramujos/metabolismo , Ácido gama-Aminobutírico/metabolismo , Animais , Autorradiografia , Cromatografia Líquida de Alta Pressão , Gânglios/citologia , Gânglios/metabolismo , Imuno-Histoquímica , Ácido gama-Aminobutírico/biossíntese , Ácido gama-Aminobutírico/imunologiaRESUMO
The pharmacological properties and modulation by lithium of the kainate (KA) receptor system coupled to the evoked release of [3H]-gamma-aminobutyric acid [( 3H]GABA) from purified populations of striatal neurons in primary culture were examined. KA evoked a dose-dependent (EC50, 100 microM) and saturable increase in [3H]GABA release from striatal neurons that was unaffected by the removal of extracellular calcium and resistant to the actions of tetrodotoxin. The release of [3H]GABA evoked by 100 microM KA was attenuated in a dose-dependent manner by the following excitatory amino acid antagonists (IC50):6-cyano-2, 3-dihydroxy-7-nitroquinoxaline (2 microM),2,3-dihydroxy-6,7-dinitroquinoxaline (2 microM), kynurenate (0.3 mM), and gamma-D-glutamylglycine (2 mM). The antagonist properties of 6-cyano-2,3-dihydroxy-7-nitroquinoxaline, kynurenate, and gamma-D-glutamylglycine were competitive in nature, inducing parallel rightward shifts of the KA dose-response curves. At concentrations at which it did not significantly increase basal levels of [3H]GABA release, quisqualate attenuated in a dose-dependent manner (IC50, 10 microM) the release due to 100 microM KA. The quisqualate receptor agonist alpha-amino-3-hydroxyisoxazolepropionic acid (AMPA), however, exerted a biphasic effect on 100 microM KA-evoked release of [3H]GABA. At lower concentrations of AMPA (0.1-10 microM), the release due to 100 microM KA was potentiated 25-50%; at higher concentrations (greater than 10 microM) AMPA induced a dose-dependent (IC50, 100 microM) attenuation of KA-evoked release. The release of [3H]GABA due to 100 microM KA was significantly potentiated by the replacement of sodium with lithium in the extracellular medium. A significant potentiation (20-30%) was detected with as little as 5-10 mM lithium, and maximal effects (100-110% increase) were obtained with 50-75 mM lithium. Replacement of sodium with choline or N-methyl-D-glucamine could not mimic the actions of lithium. Lithium (25 mM) also induced a 4-fold increase in the levels of endogenous GABA release due to 100 microM KA. Whole-cell voltage-clamp recordings of these striatal neurons indicated that the 100 microM KA-induced inward current was not significantly altered in the presence of 25 mM lithium. Lithium attenuated vasoactive intestinal polypeptide-stimulated cyclic AMP formation by 50%, with a dose dependence similar to that of its actions on KA-evoked release. The results of this study demonstrate a distinct pharmacological profile for the KA receptor system coupled to the evoked release of [3H]GABA from striatal neurons.(ABSTRACT TRUNCATED AT 400 WORDS)
Assuntos
Corpo Estriado/efeitos dos fármacos , Ácido Caínico/farmacologia , Lítio/farmacologia , Receptores de Neurotransmissores/metabolismo , Ácido gama-Aminobutírico/metabolismo , Animais , Células Cultivadas , Corpo Estriado/metabolismo , Relação Dose-Resposta a Droga , Eletrofisiologia , Camundongos , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Receptores de Ácido CaínicoRESUMO
The actions of 56 mM KCl and excitatory amino acid (EAA) agonists on the release of endogenous glycine (Gly) from striatal neurons in primary culture was examined. During a 3 min period, 2 x 10(6) striatal neurons released 743 +/- 51 pmol of Gly. In the presence of 56 mM KCl, an additional 492 +/- 52 pmol of Gly (+66%) were released, 75% of which was dependent upon the presence of extracellular calcium. When striatal neurons were exposed to 1 mM N-methyl-D-aspartate (NMDA) or quisqualate (QA), endogenous Gly released was increased by 370 +/- 71 (+50%) or 120 +/- 31 (+16%) pmol, respectively. In the presence of 1 mM kainate (KA), however, the release of endogenous Gly increased by 994 +/- 82 pmol (+135%). Interestingly, while KA (1 mM) was twice as effective as KCl (56 mM) in evoking the release of endogenous Gly, KCl was 5 times more effective than KA in evoking the release of endogenous gamma-aminobutyric acid (GABA). KA-induced increases of endogenously released Gly were dose-dependent (EC50, 100 microM), saturable and not significantly reduced in the absence of extracellular calcium. The actions of KA were blocked by coincubation with 6-cyano-2,3-dihydroxy-7-nitro-quinoxaline (CNQX), a competitive antagonist at the KA receptor. These data suggest that the release of endogenous Gly from striatal neurons in primary culture is regulated principally by EAA actions at the KA receptor system.
Assuntos
Corpo Estriado/metabolismo , Glicina/metabolismo , Ácido Caínico/farmacologia , Receptores de Superfície Celular/metabolismo , Animais , Ácido Aspártico/análogos & derivados , Ácido Aspártico/farmacologia , Células Cultivadas , Corpo Estriado/citologia , Corpo Estriado/efeitos dos fármacos , N-Metilaspartato , Oxidiazóis/farmacologia , Ácido Quisquálico , Receptores de Aminoácido , Receptores de Superfície Celular/efeitos dos fármacosRESUMO
Arterial levels of epinephrine (E) were significantly raised for all times sampled, after intracerebroventricular (i.c.v.) injection of arginine vasopressin (AVP) (1 nmol) into conscious rabbits. Venous levels of norepinephrine (NE) were significantly raised within the first 4 min after a similar i.c.v. injection of AVP. The significant increases in arterial E and venous NE correspond in time and therefore could account for changes in heart rate and blood pressure elicited by centrally injected AVP. Therefore, AVP could influence cardiovascular function by increasing the activity of the sympatho-adrenomedullary system.
Assuntos
Arginina Vasopressina/farmacologia , Encéfalo/fisiologia , Epinefrina/sangue , Norepinefrina/sangue , Animais , Pressão Sanguínea/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Injeções Intraventriculares , Masculino , CoelhosRESUMO
Previous reports that central administration of arginine vasopressin (AVP) increases turnover of brain catecholamines raise the possibility that the pressor responses which follow central administration of AVP may be mediated, in part, by central catecholamines. To test this hypothesis, rats were given intraventricular injections of vehicle, or of the neurotoxin, 6-hydroxydopamine, which resulted in significant depletions of hypothalamic and medulla oblongata noradrenalin and hypothalamic dopamine, but not of medullary dopamine or of hypothalamic and medullary 5-hydroxytryptamine. Following a one week recovery, these conscious rats, fitted with indwelling arterial catheters, were given intraventricular injections of AVP; the increases in arterial pressure and heart rate were significantly reduced in the catecholamine-depleted animals. These data support the hypothesis that the pressor and tachycardia responses to intraventricular AVP are mediated, in part, by central catecholamine-containing neurons.
Assuntos
Arginina Vasopressina/farmacologia , Pressão Sanguínea/efeitos dos fármacos , Encéfalo/fisiologia , Catecolaminas/fisiologia , Animais , Arginina Vasopressina/fisiologia , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Catecolaminas/metabolismo , Frequência Cardíaca/efeitos dos fármacos , Hidroxidopaminas , Injeções Intraventriculares , Masculino , Oxidopamina , Ratos , Ratos EndogâmicosRESUMO
The effects of mugineic acid upon excitations induced by quisqualic acid (quis), N-methyl-D-aspartic acid (NMDA) and kainic acid (kain) on rat central neurones were examined in vivo through microiontophoretic drug administration in anaesthetized rats. Mugineic acid usually caused a non-selective enhancement of drug-induced excitations, occasionally producing direct excitation itself or increasing the rate of spontaneous discharge. Mugineic acid-induced effects were not blocked by the NMDA selective antagonist, 2-amino-5-phosphonovaleric acid (AP5).
Assuntos
Aminoácidos/farmacologia , Ácido Azetidinocarboxílico/farmacologia , Azetinas/farmacologia , Neurônios/efeitos dos fármacos , Animais , Ácido Azetidinocarboxílico/análogos & derivados , Potenciais Evocados/efeitos dos fármacos , Feminino , Masculino , Neurônios/fisiologia , Plantas/análise , Ratos , Estimulação QuímicaRESUMO
The present study shows that the drug 5,7-dihydroxytryptamine (5,7-diHT) can be used reliably to deplete the neurotransmitter serotonin (5-HT) from the nervous system of the snail Helisoma. The depletion is more effective in axonal and synaptic regions (85-90%) than in the somata (55%), is reasonably specific for serotonin (dopamine is affected to a much lesser extent), and is transient, with normal levels of neurotransmitter being restored by 2 months. A physiological correlate of 5-HT depletion has been shown in that an EPSP elicited by a cerebral serotonergic neuron (C1) onto a buccal motoneuron (B19) is much smaller during depletion and also recovers with time as 5-HT regains normal concentration. Despite the severe 5-HT depletion and physiological impairment, the gross morphology of neuron C1 remains indistinguishable from controls. Serotonergic depletion is not accompanied by development of receptor supersensitivity nor by the production of serotonin in extraneuronal sources.
Assuntos
5,7-Di-Hidroxitriptamina/farmacologia , Di-Hidroxitriptaminas/farmacologia , Gânglios/análise , Serotonina/análise , Caramujos/análise , Animais , Cromatografia Líquida de Alta Pressão , Gânglios/citologia , Gânglios/fisiologia , Receptores de Serotonina/efeitos dos fármacos , Serotonina/fisiologia , Transmissão Sináptica/efeitos dos fármacosRESUMO
The effects of intrathecally administered arginine-vasopressin (AVP) and substance P (SP) on adrenal medullary secretion of epinephrine were examined in anesthetized Sprague-Dawley rats. Plasma epinephrine levels were measured in blood samples taken directly from the adrenal vein using a novel micropuncture technique. The blood samples (20-30 microliter in volume) were taken before, and 2 min, 15 min and 30 min after intrathecal injections of AVP, SP or vehicle only. Plasma was assayed for epinephrine using high pressure liquid chromatography. Adrenal venous epinephrine levels were not significantly altered by the intrathecal administration of AVP, thereby suggesting that adrenal epinephrine secretion is not involved in the cardiovascular responses previously reported to occur following similar doses of intrathecal AVP. Intrathecal SP administration, while causing blood pressure responses similar to those produced by AVP, resulted in significant increases in adrenal vein epinephrine. This finding suggests that activation of adrenal secretion of epinephrine may contribute to SP-initiated blood pressure changes.
Assuntos
Glândulas Suprarrenais/metabolismo , Arginina Vasopressina/farmacologia , Pressão Sanguínea/efeitos dos fármacos , Epinefrina/metabolismo , Substância P/farmacologia , Glândulas Suprarrenais/irrigação sanguínea , Glândulas Suprarrenais/efeitos dos fármacos , Animais , Arginina Vasopressina/administração & dosagem , Epinefrina/sangue , Injeções Espinhais , Masculino , Ratos , Ratos Endogâmicos , Substância P/administração & dosagem , VeiasRESUMO
Numerous problems have been associated with previous attempts to develop a suitable method for the induction and assessment of alcohol dependence and withdrawal syndrome in the rat. Using our modification of a common inhalation method for the long-term administration of ethanol, these problems can be eliminated. Adult male rats (Long Evans and Brattleboro) were exposed to ethanol vapor concentrations of 7 to 35 mg/liter of air, which cause rapid development of tolerance and physical dependence. With this inhalation method, it is possible to obtain and easily maintain high levels of ethanol in the blood (150 to 400 mg/dl). When exposure to ethanol is terminated, ethanol is eliminated from the system within 1 to 6 hr. This rapid elimination of ethanol is accompanied by a high susceptibility to withdrawal reactions. The severity of the withdrawal syndrome was assessed within 6 to 24 hr after cessation of the ethanol administration by exposing each rat individually to a 60 to 120-sec period of bell ringing. Convulsive seizures were observed in nearly 90% of the animals tested, with a mortality rate of less than 20%.