RESUMO
Polymorphic variants of the dopamine D(4) receptor have been consistently associated with attention-deficit hyperactivity disorder (ADHD). However, the functional significance of the risk polymorphism (variable number of tandem repeats in exon 3) is still unclear. Here, we show that whereas the most frequent 4-repeat (D(4.4)) and the 2-repeat (D(4.2)) variants form functional heteromers with the short isoform of the dopamine D(2) receptor (D(2S)), the 7-repeat risk allele (D(4.7)) does not. D(2) receptor activation in the D(2S)-D(4) receptor heteromer potentiates D(4) receptor-mediated MAPK signaling in transfected cells and in the striatum, which did not occur in cells expressing D(4.7) or in the striatum of knockin mutant mice carrying the 7 repeats of the human D(4.7) in the third intracellular loop of the D(4) receptor. In the striatum, D(4) receptors are localized in corticostriatal glutamatergic terminals, where they selectively modulate glutamatergic neurotransmission by interacting with D(2S) receptors. This interaction shows the same qualitative characteristics than the D(2S)-D(4) receptor heteromer-mediated mitogen-activated protein kinase (MAPK) signaling and D(2S) receptor activation potentiates D(4) receptor-mediated inhibition of striatal glutamate release. It is therefore postulated that dysfunctional D(2S)-D(4.7) heteromers may impair presynaptic dopaminergic control of corticostriatal glutamatergic neurotransmission and explain functional deficits associated with ADHD.
Assuntos
Transtorno do Deficit de Atenção com Hiperatividade/genética , Multimerização Proteica , Receptores de Dopamina D2/metabolismo , Receptores de Dopamina D4/metabolismo , Animais , Células CHO , Corpo Estriado/metabolismo , Cricetinae , Técnicas de Introdução de Genes/métodos , Ácido Glutâmico/metabolismo , Células HEK293 , Humanos , Técnicas In Vitro , Camundongos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Neurônios/metabolismo , Transdução de Sinais , Transfecção/métodosRESUMO
Monoamines such as dopamine, histamine and serotonin (5-HT) are widely distributed throughout the brain of the fruit fly Drosophila melanogaster, where many of their actions have been investigated. For example, histamine is released from photoreceptor synapses in the lamina neuropile of the visual system. Mutations of the genes white, an important eye pigmentation marker in fly genetics that encodes an ABC transporter, and its binding partner brown, cause neural phenotypes not readily reconciled solely with actions in eye pigmentation. We find that flies mutant for these genes, and another binding partner, scarlet, have about half the wild-type amount of histamine in the head, as well as reduced 5-HT and dopamine. These differences parallel reductions in immunoreactivity to the corresponding biogenic amines. They also correlate with the amine content of fractions after differential centrifugation of head homogenates. Thus, most of the amine is found in the vesicle-rich fraction of wild-type head homogenates, whereas it is found in the supernatant fractions from white, brown and scarlet flies. White co-expresses in lamina epithelial glia with Ebony, which conjugates histamine to beta-alanine. Histamine is then released when the conjugate is hydrolyzed in photoreceptors, by Tan. Mutant white ameliorates the effects of tan on head histamine whereas it exacerbates the effects of ebony. Our results are consistent with the proposal that histamine uptake by the epithelial glia may be white dependent. Behavioral abnormalities in white, brown and scarlet mutants could arise because aminergic neurons in the Drosophila brain have reduced amine for release.
Assuntos
Transportadores de Cassetes de Ligação de ATP/fisiologia , Encéfalo/fisiologia , Proteínas de Drosophila/fisiologia , Drosophila melanogaster/fisiologia , Proteínas do Olho/fisiologia , Histamina/análise , Transportadores de Cassetes de Ligação de ATP/genética , Análise de Variância , Animais , Cromatografia Líquida de Alta Pressão , Olho Composto de Artrópodes/fisiologia , Proteínas de Drosophila/genética , Drosophila melanogaster/citologia , Drosophila melanogaster/genética , Epitélio/fisiologia , Proteínas do Olho/genética , Genes de Insetos , Genótipo , Cabeça/fisiologia , Mutação , Neurópilo/fisiologia , Neurotransmissores/genética , Neurotransmissores/fisiologia , FenótipoRESUMO
Histamine, a ubiquitous aminergic messenger throughout the body, also serves as a neurotransmitter in both vertebrates and invertebrates. In particular, the photoreceptors of adult arthropods use histamine, modulating its release to signal increases and decreases in light intensity. Strong evidence from various arthropod species indicates that histamine is synthesized and stored in photoreceptors, undergoes Ca-dependent release, inhibits postsynaptic interneurons by gating Cl channels, and is then recycled. In Drosophila, the synthetic enzyme, histidine decarboxylase, and the subunits of the histamine-gated chloride channel have been cloned. Possible histamine transporters at synaptic vesicles and for reuptake remain elusive. Indeed, the mechanisms that remove histamine from the synaptic cleft, and that help terminate histamine's action, are unexpectedly complex, their details remaining unresolved. A major pathway in Drosophila, and possibly other arthropod species, is by conjugation of histamine to beta-alanine to form carcinine in adjacent glia. This conjugate then returns to the photoreceptors where it is hydrolysed to liberate histamine, which is then loaded into synaptic vesicles. Evidence from other species suggests that direct reuptake of histamine into the photoreceptors may also occur. Light depolarizes the photoreceptors, causing histamine release and postsynaptic inhibition; dimming hyperpolarizes the photoreceptors, causing a decrease in histamine release and an "off" response in the postsynaptic cell. Further pursuit of histamine's action at these highly specialized synapses should lead to an understanding of how they signal minute changes in presynaptic membrane potential, how they reliably extract signals from noise, and how they adapt to a wide range of presynaptic membrane potentials.
Assuntos
Artrópodes/anatomia & histologia , Histamina/metabolismo , Células Fotorreceptoras de Invertebrados/fisiologia , Transdução de Sinais/fisiologia , Sinapses/fisiologia , Animais , Artrópodes/fisiologiaRESUMO
Reliable estimates of the quantum size in histaminergic neurons are not available. We have exploited two unusual opportunities in the fly's (Drosophila melanogaster) visual system to make such determinations for histaminergic photoreceptor synapses: 1) the possibility to microdissect successively from whole fly heads freeze-dried in acetone: the compound eyes; the first optic neuropils, or lamina; and the rest of the brain; and 2) the uniform sheaves of lamina synaptic terminals of photoreceptors R1-R6. We used this organization to count scrupulously the numbers of 30-nm synaptic vesicles from electron micrographs of R1-R6 profiles, and from microdissections we determined the regional contents of histamine in the compound eye, lamina, and central brain. Total head histamine averages 1.98 ng of which 9% was lost after freeze-drying in acetone and a further 28% after the brain was microdissected. Of the remainder, 71% was in the eye and lamina. Assuming that histamine loss from the tissue occurred mostly by diffusion evenly distributed among all regions, the overall lamina content of the head would be 0.1935 ng before dissection. From published values for the volumes of the brain's compartments, the computed regional concentrations of histamine are highest in the lamina (4.35 mM) because of the terminals of R1-R6. The concentration in the retina is approximately 13% that in the lamina, suggesting that most histamine is vesicular. There are approximately 43,500 +/- 7,400 (SD) synaptic vesicles per terminal and, if all histamine is allocated equally and exclusively among these, the vesicle contents would be 858 +/- 304 x 10(-21) moles or approximately 5,000 +/- 1,800 (SD) molecules at an approximate concentration of 670 mM. These values are compared with the vesicle contents at synapses using acetylcholine and catecholamines.
Assuntos
Encéfalo/metabolismo , Histamina/metabolismo , Células Fotorreceptoras de Invertebrados/citologia , Sinapses/metabolismo , Análise de Variância , Animais , Encéfalo/ultraestrutura , Cromatografia Líquida de Alta Pressão/métodos , Drosophila melanogaster , Técnica de Fratura por Congelamento/métodos , Cabeça/anatomia & histologia , Cabeça/fisiologia , Imuno-Histoquímica/métodos , Microdissecção/métodos , Microscopia Eletrônica de Transmissão/métodos , Células Fotorreceptoras de Invertebrados/metabolismo , Sinapses/ultraestrutura , Vesículas Sinápticas/metabolismo , Vesículas Sinápticas/ultraestruturaRESUMO
In the fly's visual system, two classes of lamina interneuron, L1 and L2, cyclically change both their size and shape in a rhythm that is circadian. Several neurotransmitters and the lamina's glial cells are known to be involved in regulating these rhythms. Moreover, vacuolar-type H+-ATPase (V-ATPase) in the optic lobe is thought also to participate in such regulation. We have detected V-ATPase-like immunoreactivity in the heads of both Drosophilla melanogaster and Musca domestica using antibodies raised against either the B- or H-subunits of V-ATPase from D. melanogaster or against the B-subunit from two other insect species Culex quinquefasciatus and Manduca sexta. In the visual systems of both fly species V-ATPase was localized immunocytochemically to the compound eye photoreceptors. In D. melanogaster immunoreactivity oscillated during the day and night and under constant darkness the signal was stronger during the subjective night than the subjective day. In turn, blocking V-ATPase by injecting a V-ATPase blocker, bafilomycin, in M. domestica increased the axon sizes of L1 and L2, but only when bafilomycin was applied during the night. As a result bafilomycin abolished the day/night difference in axon size in L1 and L2, their sizes being similar during the day and night.
Assuntos
Dípteros/enzimologia , Olho/citologia , Lobo Óptico de Animais não Mamíferos/enzimologia , ATPases Vacuolares Próton-Translocadoras/metabolismo , Animais , Tamanho Celular , Culex/enzimologia , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/enzimologia , Proteínas de Insetos/metabolismo , Manduca/enzimologia , Lobo Óptico de Animais não Mamíferos/citologiaRESUMO
Photoreceptors are richly supplied with mitochondria, where they are required to meet the energetic demands, in the soma, of phototransduction and, in the terminal, of neurotransmitter release. Compromising the latter, we have made photoreceptors R1-R6 in Drosophila ommatidia homozygous for either of two alleles, milt(186) and milt(92), of milton in whole-eye mosaics. Such mutant photoreceptors fail to target mitochondria to their terminals. We show from quantitative electron microscopy (EM) that mitochondria are totally lacking at the terminal but nevertheless abundant and present throughout the soma, where their distribution differs from that of control ommatidia, however, being more heavily concentrated in the nuclear region. Mitochondria are sparse at the basalmost level of mutant ommatidia, and are lacking beneath the basement membrane, in the axons and terminals of these cells. The absence of mitochondria from R1-R6 terminals and concommitant reductions in synaptic vesicle packing density, previously reported, we show here are accompanied by reduced immunoreactivity to the photoreceptor transmitter histamine but not by any change in total head histamine content, as determined by high-performance liquid chromatography. Mutant terminals also contain vesicle profiles with a wider range of sizes. These two phenotypes suggest that the reduced availability of ATP when mutant terminals lack a mitochondrial supply compromises their ability to pump histamine into synaptic vesicles and perturbs membrane distribution within the terminal. In addition, a band of somata in the lamina cortex, at least some of which are postsynaptic neurons not homozygous for milton, also shows altered mitochondrial targeting, with abnormal clusters of mitochondria, as visualized by immunolabeling with anti-hsp and by serial EM. Within the lamina, terminals of mutant photoreceptors are penetrated by neighboring cells with invaginations that frequently contain mitochondria, suggesting that a mechanism exists for intercellular metabolic support. Our findings indicate the direct and compensatory responses in a population of neurons when mitochondria are not correctly targeted to their synaptic terminals.
Assuntos
Proteínas de Drosophila/metabolismo , Mitocôndrias/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Células Fotorreceptoras/metabolismo , Animais , Cromatografia Líquida , Drosophila , Proteínas de Drosophila/deficiência , Imuno-Histoquímica , Microscopia Eletrônica , Proteínas do Tecido Nervoso/deficiênciaRESUMO
The effect of acute and repeated (once daily, 14 days) administration of a potential antidepressant, the glycineB partial agonist 1-aminocyclopropanecarboxylic acid (ACPC, 100-400 mg/kg, ip), on the hyperactivity induced by amphetamine (0.5 mg/kg, sc) in rats was studied. Neither acute nor repeated treatment with the drug affected the hyperlocomotion induced by amphetamine. The obtained results indicate that ACPC does not resemble antidepressant drugs in this behavioral model.
Assuntos
Aminoácidos Cíclicos/farmacologia , Anfetamina/toxicidade , Hipercinese/induzido quimicamente , Receptores de N-Metil-D-Aspartato/antagonistas & inibidores , Anfetamina/antagonistas & inibidores , Análise de Variância , Animais , Relação Dose-Resposta a Droga , Locomoção/efeitos dos fármacos , Masculino , Ratos , Ratos Wistar , Receptores de Glicina/agonistasRESUMO
Using the conflict drinking Vogel test in rats as a model we examined the anxiolytic-like activity of (S)-4-carboxyphenylglycine (S-4CPG), an antagonist of group I metabotropic glutamate receptors (mGlu receptors), of (RS)-a-methylserine-O-phosphate-monophenyl ester (MSOPPE), an antagonist of group II mGlu receptors, and of (2S,1'S,2'S)-2-(carboxycyclopropyl)glycine (L-CCG-I), an agonist of group II mGlu receptors. The obtained results indicate that intrahippocampal administration of S-4CPG and L-CCG-I, but not MSOPPE to rats produces a dose-dependent anticonflict effect, which is unrelated to the reduced perception of the stimulus or to an increased thirst drive. The hippocampus may be one of the neuroanatomical sites of the anxiolytic-like effects of either agent.
Assuntos
Ansiolíticos/farmacologia , Receptores de Glutamato Metabotrópico/efeitos dos fármacos , Animais , Comportamento Animal , Benzoatos/farmacologia , Conflito Psicológico , Comportamento de Ingestão de Líquido , Glicina/análogos & derivados , Glicina/farmacologia , Masculino , Fosfosserina/farmacologia , Ratos , Ratos Wistar , Receptores de Glutamato Metabotrópico/agonistas , Receptores de Glutamato Metabotrópico/antagonistas & inibidoresRESUMO
Histamine is a neurotransmitter at arthropod photoreceptors. Even though the fruit fly, Drosophila melanogaster, is a widely used model in neuroscience research, the histamine content of its nervous system has not so far been reported. We have developed a high performance liquid chromatography (HPLC) method with pre-column o-phtaldialdehyde-mercaptoethanol (OPA-ME) derivatization and electrochemical detection, to determine this amine in Drosophila. The histamine content of the fly's head averages about 2.0 ng per head. In heads of the mutant hdc(JK910), a presumed null for the gene encoding the enzyme that synthesizes histamine, histamine was not detected in measurable amounts. In heads of the mutant sine oculis, which lacks compound eyes, only 28% of this amine was found compared with wild type flies, so histamine is mainly present in the compound eye photoreceptors. Also observed in histamine-deficient mutants was a decrease in the peak which contains a substance having the same retention time as carcinine (beta-alanyl-histamine). Our method was not able to detect compounds previously reported as histamine metabolites in insects. In spite of this, the method we have developed enables the fast and accurate measurement of histamine in the heads of Drosophila, suitable for screening mutants.
Assuntos
Drosophila/química , Cabeça/fisiologia , Histamina/análise , Sistema Nervoso/química , Animais , Cromatografia Líquida de Alta Pressão , Drosophila/citologia , Drosophila/metabolismo , Cabeça/anatomia & histologia , Histamina/análogos & derivados , Histamina/metabolismo , Mercaptoetanol/química , Sistema Nervoso/citologia , Sistema Nervoso/metabolismo , Neurônios/química , Neurônios/citologia , Neurônios/metabolismo , Células Fotorreceptoras/química , Células Fotorreceptoras/citologia , Células Fotorreceptoras/metabolismo , o-Ftalaldeído/químicaRESUMO
Three series of new 9-substituted 1,2,3,4-tetrahydro-beta-carbolin-1-ones with 2-, 3- and 4-membered alkyl chain (1, 2 and 3, respectively) were synthesized, and the effect of some structural modifications on their 5-HT1A and 5-HT2A receptor affinities and functional in vivo properties was discussed. Radioligand binding measurements showed that the majority of compounds had a distinct affinity for 5-HT1A (1b, 2a, 2b, 2c, 3b; Ki = 0.3-64 nM) and 5-HT2A receptors (1b, 2b, 2c, 3b; Ki = 0.9-80 nM). The most potent 5-HT1A (1b, 2a, 2b, 3b) and 5-HT2A (1b, 2b, 3b) ligands were evaluated in in vivo tests. The obtained results indicate that 1,2,3,4-tetrahydro-beta-carbolin-1-ones containing 1-(o-methoxyphenyl)piperazine (1-3b) show pharmacological profile of 5-HT1A postsynaptic antagonists (with very weak agonistic component) and 5-HT2A antagonists, compound with 1,2,3,4-tetrahydroisoquinoline (2a) is a pure 5-HT1A postsynaptic antagonist. Summing up, the connection of 1,2,3,4-tetrahydro-beta-carbolin-1-one moiety through the 2-4-membered alkyl spacer with 1-(o-methoxyphenyl)-piperazine, which is present in a variety of 5-HT1A ligands, allowed us to obtain the compounds with high and equal affinity for 5-HT1A/5-HT2A receptors and the expected functional properties, i.e. distinct antagonistic and weak agonistic activity at 5-HT1A postsynaptic receptors and antagonistic at 5-HT2A ones.
Assuntos
Carbolinas/metabolismo , Carbolinas/farmacologia , Receptores de Serotonina/metabolismo , Antagonistas da Serotonina/metabolismo , Antagonistas da Serotonina/farmacologia , Agonistas do Receptor de Serotonina/metabolismo , Agonistas do Receptor de Serotonina/farmacologia , 8-Hidroxi-2-(di-n-propilamino)tetralina/farmacologia , Inibidores da Captação Adrenérgica/farmacologia , Animais , Comportamento Animal/efeitos dos fármacos , Córtex Cerebral/metabolismo , Hipocampo/metabolismo , Ligantes , Masculino , Ensaio Radioligante , Ratos , Ratos Wistar , Receptor 5-HT2A de Serotonina , Receptores 5-HT1 de Serotonina , Reserpina/farmacologia , Relação Estrutura-AtividadeRESUMO
The purpose of the present study was to assess the activity of (+)-oxaprotiline [(+)-OXA] (a noradrenaline uptake inhibitor) and (-)-oxaprotiline [(-)-OXA] (with unknown mechanism of action) in two experimental models of pain in mice, a hot plate test and a writhing syndrome induced by phenylbenzoquinone (PHBQ), and to determine whether the opioidergic system may be engaged in their antinociceptive effects. Morphine was used as a reference drug. Administration of (+)-OXA (0.31-5 mg/kg) and (-)-OXA (20 mg/kg) produced a statistically significant elevation of the nociceptive threshold, measured by the increased latencies in the hot plate test. Moreover, (+)-OXA (0.62-5 mg/kg) and (-)-enantiomer (5-20 mg/kg) decreased the number of writhing episodes induced by PHBQ in mice, (+)-enantiomer being more effective than (-)-OXA in either test. In the hot plate test, the analgesic effect induced by (+)-OXA (0.31 mg/kg) or (-)-OXA (20 mg/kg) was abolished by naloxone (2 mg/kg), an opioid receptor antagonist. In the writhing test, naloxone (2 mg/kg) partially, but not significantly, reduced the antinociceptive responses induced by (+)-OXA (0.62 mg/kg) or (-)-OXA (5 mg/kg). The obtained results show that both OXA enantiomers produce antinociception in mice which can be, at least partially, connected with opioid system.
Assuntos
Antidepressivos/farmacologia , Maprotilina/análogos & derivados , Nociceptores/efeitos dos fármacos , Dor/tratamento farmacológico , Animais , Benzoquinonas/farmacologia , Modelos Animais de Doenças , Masculino , Maprotilina/farmacologia , Camundongos , Naloxona/farmacologia , Antagonistas de Entorpecentes/farmacologia , Medição da Dor/efeitos dos fármacos , EstereoisomerismoRESUMO
Structural modifications of 1, a postsynaptic 5-HT(1A) receptor antagonist, provided its flexible (8, 12) and rigid (7, 9, 11, 13) analogues. Compounds 7, 8, 9, and 11 showed high 5-HT(1A) receptor affinity (K(i) = 4-72 nM). They acted as 5-HT(1A) postsynaptic receptor antagonists, since, like 1, they inhibited the behavioral syndrome, i.e., flat body posture (FBP) and forepaw treading (FT), in reserpine-pretreated rats as well as the lower lip retraction (LLR) in rats, both induced by 8-hydroxy-2-(di-n-propylamino)tetralin hydrobromide (8-OH-DPAT), a 5-HT(1A) receptor agonist. Compound 12, which demonstrated high 5-HT(1A) receptor affinity (K(i) = 50 nM), revealed properties of a partial 5-HT(1A) receptor agonist: it induced LLR and, at the same time, inhibited FT in rats. Compound 13 (K(i) = 1600 nM) was not tested in a behavioral study. Restriction of the conformational freedom in 2, a full 5-HT(1A) receptor antagonist, yielded compound 14 with high 5-HT(1A) receptor affinity (K(i) = 47 nM) and partial agonist properties at postsynaptic 5-HT(1A) receptors in the above tests in vivo; i.e., it induced LLR and inhibited FBP and FT in rats. New constrained analogues of 1 and 2 (compounds 7 and 14, respectively) were also synthesized to recognize a bioactive conformation of those 5-HT(1A) receptor antagonists. On the basis of in vitro and in vivo investigations, binding and functional properties of compound 7 were found to reflect those of 1 at 5-HT(1A) receptors. On the other hand, compound 14, a rigid analogue of 2, showed a different activity in vivo in comparison with the parent compound. PM3 and MM calculations revealed the existence of three low-energy conformers of 7 and six of 14, all of them belonging to the extended family of conformations. The optimized structures of both analogues had a different angle between aromatic planes of terminal fragments; moreover, the heteroaromatic system of those molecules occupied various space regions. Our present study provides support to the hypothesis that the bioactive conformation of 1, responsible for its postsynaptic 5-HT(1A) receptor antagonism, is an extended linear structure represented by 7.
Assuntos
Piperazinas/química , Receptores de Serotonina/efeitos dos fármacos , Antagonistas da Serotonina/química , Triazóis/química , Animais , Comportamento Animal/efeitos dos fármacos , Lábio , Masculino , Modelos Moleculares , Conformação Molecular , Estrutura Molecular , Atividade Motora , Ftalimidas/síntese química , Ftalimidas/metabolismo , Ftalimidas/farmacologia , Piperazinas/síntese química , Piperazinas/metabolismo , Piperazinas/farmacologia , Postura , Ratos , Ratos Wistar , Receptores de Serotonina/metabolismo , Receptores 5-HT1 de Serotonina , Antagonistas da Serotonina/metabolismo , Relação Estrutura-Atividade , Triazóis/síntese química , Triazóis/metabolismo , Triazóis/farmacologiaRESUMO
It has been suggested that adrenergic agents might modulate the L-arginine-NO pathway. Sympathomimetic agonists enhance the basal release of NO, and noradrenaline increases the synthesis of nitric oxide synthase (NOS) in the medial basal hypothalamus in vitro. In the present study possible involvement of NO in central stimulation of the hypothalamic-pituitary-adrenal (HPA) axis by adrenergic agents was investigated in conscious rats. The nitric oxide synthase blocker N(omega)-nitro-L-arginine methyl ester (L-NAME 2 and 10 microg) was administered intracerebroventricularly (i.c.v.) 15 min before the adrenergic agonist given by the same route; 1 h later the rats were decapitated. Plasma levels of ACTH and corticosterone were measured. L-NAME significantly diminished the ACTH and corticosterone response to phenylephrine (30 microg), an alpha1-adrenergic receptor agonist. These hormone responses to clonidine (10 microg), an alpha2-receptor agonist, were dose-dependently suppressed or totally abolished by L-NAME. A significant rise in the ACTH and corticosterone secretion induced by isoprenaline (10 microg), a beta-adrenergic receptor agonist, was only moderately diminished by pretreatment with L-NAME. These results indicate that NOS is considerably involved in central stimulation of the HPA axis by alpha1- and alpha2-adrenergic receptor agonists, and that NO mediates the stimulatory action of these agonists on ACTH and corticosterone secretion. The stimulation induced by beta-adrenergic receptors is only moderately affected by endogenous NO.
Assuntos
Agonistas Adrenérgicos/administração & dosagem , Hormônio Adrenocorticotrópico/sangue , Corticosterona/sangue , Inibidores Enzimáticos/farmacologia , Sistema Hipotálamo-Hipofisário/efeitos dos fármacos , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico Sintase/antagonistas & inibidores , Sistema Hipófise-Suprarrenal/efeitos dos fármacos , Hormônio Adrenocorticotrópico/efeitos dos fármacos , Animais , Clonidina/farmacologia , Isoproterenol/farmacologia , Masculino , Óxido Nítrico , Óxido Nítrico Sintase/fisiologia , Óxido Nítrico Sintase Tipo I , Fenilefrina/farmacologia , Ratos , Ratos Wistar , Receptores Adrenérgicos/efeitos dos fármacosRESUMO
Putative involvement of endogenous nitric oxide (NO) in the corticotropin-releasing hormone (CRH, 1 microg/kg i.p.)- and vasopressin (AVP, 5 microg/kg i.p.)-induced ACTH and corticosterone secretion was investigated in both non-stressed and crowded rats. The NO synthase blocker Nomega-nitro-l-arginine (l-NNA, 2 mg/kg i.p. ) significantly augmented the AVP-induced ACTH and corticosterone secretion in control and stressed rats, but it increased the CRH-induced ACTH response only in control rats. Crowding stress did not affect the l-NNA evoked increase in AVP-induced hormone responses, but it abolished the CRH-induced ACTH response.
Assuntos
Arginina Vasopressina/metabolismo , Hormônio Liberador da Corticotropina/metabolismo , Óxido Nítrico/fisiologia , Sistema Hipófise-Suprarrenal/fisiologia , Meio Social , Estresse Psicológico/fisiopatologia , Animais , Inibidores Enzimáticos/farmacologia , Masculino , Óxido Nítrico Sintase/antagonistas & inibidores , Nitroarginina/farmacologia , Ratos , Ratos Wistar , Taxa SecretóriaRESUMO
Immunomodulation of cell-mediated immunity is demonstrated in mice, after administration of desipramine, a noradrenaline-reuptake inhibitor, with or without exposing the mice later to an acute swimming stress. A single i.p. injection of 10 mg/kg desipramine to naive mice increased the relative weight of their spleens, the response of their splenocytes to the mitogen concavaline-A and their ability to produce IL-10, as compared to saline controls. Exposing the desipramine-treated mice to a swimming stress significantly reduced these parameters, as well as the levels of IL-2 and IFN-gamma, as compared to desipramine-treated mice. Stress alone reduced the weight of the spleen, and the ability of splenocytes to produce IFN-gamma. As desipramine and acute stress have stimulatory effect on the sympathetic system, it is suggested that a concomitant administration of the drug and a stressful event of these mice, change the splenocytes' micro-environment of sympathetic transmitters, and inhibit their function. These results may be partially due to impairment in the T-helper cell function by a beta-adrenoreceptor-dependent mechanism.
Assuntos
Antidepressivos Tricíclicos/farmacologia , Desipramina/farmacologia , Estresse Fisiológico/imunologia , Animais , Concanavalina A/farmacologia , Citocinas/biossíntese , Ativação Linfocitária/efeitos dos fármacos , Linfocinas/biossíntese , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mitógenos/farmacologia , Tamanho do Órgão/efeitos dos fármacos , Baço/anatomia & histologia , Baço/citologia , Baço/imunologia , Natação , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , Sais de Tetrazólio , Tiazóis , Timo/anatomia & histologiaRESUMO
Involvement of the central adrenergic system in stimulation of the hypothalamic-pituitary-adrenal (HPA) axis by carbachol, a cholinergic muscarinic agonist, was assessed indirectly through corticosterone secretion. Carbachol (2 micrograms) given intracerebroventricularly or intraperitoneally evoked a dose-related increase in serum corticosterone levels. On a molar basis, carbachol given i.c.v. was considerably more active than when injected i.p., indicating its central site of action. The corticosterone response to i.c.v. carbachol was significantly reduced by pretreatment of rats 15 min earlier with prazosin, an alpha 1-adrenergic receptor antagonist. Pretreatment with yohimbine, an alpha 2-adrenergic antagonists, did not significantly affect the carbachol-induced corticosterone response. Propranolol, a beta-adrenergic blocker, given i.c.v. or i.p. significantly impaired the carbachol-elicited corticosterone secretion. The selective noradrenergic neurotoxin DSP-4 (50 mg/kg) given i.p. 8 days before the experiment, also potently diminished the carbachol-induced rise in serum corticosterone levels. Carbachol markedly increased, while DSP-4 significantly diminished the hypothalamic noradrenaline levels. Likewise, DSP-4 significantly impaired the carbachol-induced rise in hypothalamic noradrenaline levels. Our present results indicate that the central adrenergic system is involved in the cholinergic muscarinic stimulation of the pituitary-adrenocortical response. Both hypothalamic noradrenaline and adrenergic alpha 1- and beta-receptors are significantly involved in the carbachol-induced HPA response.
Assuntos
Carbacol/farmacologia , Agonistas Muscarínicos/farmacologia , Agonistas Nicotínicos/farmacologia , Sistema Hipófise-Suprarrenal/efeitos dos fármacos , Receptores Adrenérgicos/fisiologia , Antagonistas Adrenérgicos/farmacologia , Animais , Benzilaminas/farmacologia , Corticosterona/sangue , Dopamina/metabolismo , Hipotálamo/efeitos dos fármacos , Hipotálamo/fisiologia , Injeções Intraventriculares , Masculino , Norepinefrina/metabolismo , Sistema Hipófise-Suprarrenal/metabolismo , Ratos , Ratos Wistar , Receptores Adrenérgicos/efeitos dos fármacosRESUMO
The effect of nicotine on ACTH and corticosterone secretion and possible mediation of prostaglandins in this secretion was investigated in conscious rats. Nicotine (5 and 10 mg/kg i.p.) considerably increased the plasma ACTH and corticosterone levels, measured 1h after injection. Mecamylamine (10 and 50 micrograms i.c.v.), a nicotinic receptor antagonist, given 15 min prior to nicotine dose-dependently diminished the ACTH and corticosterone responses, by 59 and 30% respectively. Pretreatment with hexamethonium (2 mg/kg i.p.), a peripheral blocker of nicotinic receptors, diminished to a similar extent the nicotine-induced ACTH and corticosterone responses. On the other hand atropine, a muscarinic receptor antagonist, did not markedly alter those responses. Systemic or intracerebroventricular pretreatment with indomethacin (2 mg/kg i.p. or 0.1 and 1 microgram i.c.v.), a cyclooxygenase and endogenous prostaglandin synthesis blocker considerably reduced, by 58%, the nicotine-induced ACTH response, but did not alter the corticosterone response. These results show that nicotine given systemically stimulates ACTH and corticosterone secretion by selective activation of central and peripheral acetylcholine nicotinic receptors. Endogenous prostaglandins are significantly involved in the nicotine-induced central stimulation of ACTH secretion. Prostaglandins do not directly affect the nicotine-induced corticosterone secretion from the adrenal cortex.
Assuntos
Hormônio Adrenocorticotrópico/sangue , Corticosterona/sangue , Inibidores de Ciclo-Oxigenase/farmacologia , Indometacina/farmacologia , Nicotina/farmacologia , Animais , Hexametônio/farmacologia , Masculino , Mecamilamina/farmacologia , Antagonistas Nicotínicos/farmacologia , Ratos , Ratos WistarRESUMO
This study was designed to determine the role of endogenous nitric oxide (NO) in the corticotropin-releasing hormone (CRH)-induced ACTH and corticosterone secretion, as well as possible involvement of hypothalamic dopamine and noradrenaline in that secretion in conscious rats. CRH given i.p. stimulated dose-dependently the pituitary-adrenocortical activity measured 1 h later. Dexamethasone (0.2 mg/kg i.p.) injected 1 h before CRH (1 microg/kg i.p.) totally abolished the CRH-elicited ACTH and corticosterone secretion, indicating a predominantly pituitary site of CRH-evoked stimulation. L-arginine (120 mg/kg i.p.) and N(omega)-nitro-L-arginine methyl ester (L-NAME 5-10 mg/kg i.p.) did not markedly affect the basal plasma ACTH and corticosterone levels. L-NAME given 15 min before CRH markedly, but not significantly, augmented the CRH-induced ACTH response, and enhanced more potently and significantly the corticosterone response. Pretreatment with L-arginine, a substrate for NOS, slightly diminished the CRH-induced ACTH response and considerably reduced the corticosterone response. L-arginine also significantly reversed the L-NAME-evoked increase in the CRH-induced ACTH and corticosterone secretion. L-NAME did not markedly alter the CRH-induced hypothalamic dopamine and noradrenaline levels, while L-arginine significantly increased noradrenaline level. However, those alterations were not directly correlated with the observed changes in ACTH and corticosterone secretion. These results indicate that in conscious rats NO plays a marked inhibitory role in the CRH-induced ACTH secretion and inhibits more potently corticosterone secretion. Hypothalamic dopamine and noradrenaline do not seem to be directly involved in the observed alterations in ACTH and corticosterone secretion.
Assuntos
Hormônio Adrenocorticotrópico/metabolismo , Corticosterona/metabolismo , Hormônio Liberador da Corticotropina/farmacologia , Inibidores Enzimáticos/farmacologia , NG-Nitroarginina Metil Éster/farmacologia , Animais , Arginina/farmacologia , Catecolaminas/metabolismo , Corticosterona/antagonistas & inibidores , Dexametasona/antagonistas & inibidores , Dexametasona/farmacologia , Sinergismo Farmacológico , Glucocorticoides/farmacologia , Hipotálamo/efeitos dos fármacos , Hipotálamo/metabolismo , Masculino , Óxido Nítrico Sintase/antagonistas & inibidores , RatosRESUMO
In the present study the role of endogenous nitric oxide (NO) in the vasopressin-induced ACTH and corticosterone secretion was investigated in conscious rats. Vasopressin (AVP 5 microg/kg i.p.) considerably augmented ACTH and corticosterone secretion. L-arginine (120 and 300 mg/kg i.p.) did not significantly alter the AVP-induced secretion of those hormones. Nitric oxide synthase (NOS) blockers N(omega)-nitro-L-arginine (L-NNA) and its methyl ester (L-NAME) given i.p. 15 min before AVP markedly increased the AVP-induced ACTH secretion. L-NNA (2 mg/kg) more potently and significantly increased the AVP-induced ACTH secretion, whereas L-NAME elicited a weaker and not significant effect. Both those NOS antagonists intensified significantly and to a similar extent the AVP-induced corticosterone secretion. L-arginine (120 mg/kg i.p.) reversed the L-NNA-induced rise in the AVP-stimulated ACTH secretion and substantially diminished the accompanying corticosterone secretion. Neither vasopressin alone nor in combination with L-arginine and L-NAME evoked any significant alterations in the hypothalamic noradrenaline and dopamine levels. L-NNA (2 and 10 mg/kg i.p.) elicited a dose dependent and significant decrease in the hypothalamic noradrenaline level. The hypothalamic dopamine level was not significantly altered by any treatment. These results indicate that in conscious rats endogenous NO has an inhibitory influence on the AVP-induced increase in ACTH and corticosterone secretion. L-NNA is significantly more potent than L-NAME in increasing the AVP-induced ACTH secretion. This may be connected with a considerable increase by L-NNA of hypothalamic noradrenergic system activation which stimulates the pituitary-adrenal axis in addition to specific inhibition of NOS.
Assuntos
Arginina Vasopressina/farmacologia , Catecolaminas/metabolismo , Inibidores Enzimáticos/farmacologia , Hipotálamo/metabolismo , NG-Nitroarginina Metil Éster/farmacologia , Nitroarginina/farmacologia , Sistema Hipófise-Suprarrenal/efeitos dos fármacos , Hormônio Adrenocorticotrópico/metabolismo , Animais , Corticosterona/metabolismo , Hipotálamo/efeitos dos fármacos , Masculino , Óxido Nítrico Sintase/antagonistas & inibidores , Sistema Hipófise-Suprarrenal/fisiologia , Ratos , Ratos WistarRESUMO
Nitric oxide synthase, an enzyme responsible for nitric oxide (NO) formation has been found in the hypothalamic paraventricular nucleus and median eminence, structures closely associated with regulation of the pituitary activity, and the pituitary gland itself. Nitric oxide modulates the stimulated release of CRH from the rat hypothalamus in vitro, which suggests its role in regulating the secretion of ACTH from the pituitary corticotrops and of corticosterone from the adrenal cortex. The purpose of the present study was to elucidate the yet unknown role of endogenous NO in the HPA response to central cholinergic stimulation in conscious rats. Neither L-arginine an NO precursor, nor the NO synthase blockers N omega-nitro-L-arginine methyl ester (L-NAME) and N omega-nitro-L-arginine (L-NNA) caused any consistent changes in the basal serum corticosterone levels. L-arginine, given in higher doses (120-150 mg/kg ip) 15 min prior to icv carbachol (2 micrograms), markedly diminished the carbachol-induced rise in corticosterone secretion. Systemic pretreatment with the nitric oxide synthase inhibitor L-NAME (5 mg/kg) significantly raised the carbachol-elicited corticosterone response, while addition of L-arginine completely blocked the effect of L-NAME. A similar increase in the carbachol-induced corticosterone response was produced by icv pretreatment with L-NAME (2 micrograms), indicating a central site of the NO interaction with cholinergic stimulation of the HPA response. L-NAME is a weak inhibitor of neuronal NOS itself, and must first be de-estrified to N omega-nitro-L-arginine to potently inhibit this enzyme. Systemic (10 mg/kg) and icv (1 microgram) pretreatment with L-NNA enhanced more effectively the carbachol-induced rise in corticosterone secretion than did pretreatment with L-NAME by either route. These results are the first direct evidence that endogenous NO significantly inhibits the HPA response to central cholinergic, muscarinic receptor stimulation under in vivo conditions.