RESUMEN
Background: Orexin-A (OrxA) administration in the posterior paraventricular nucleus of the thalamus (pPVT) reinstates extinguished cocaine-seeking behaviour following extended access to the drug (a model of dependence). The pPVT receives and integrates information associated with emotionally salient events and sends excitatory inputs to brain regions involved in the expression of emotional states, such as those driving cocaine-seeking behaviour (i.e., the nucleus accumbens, the central nucleus of the amygdala [CeA], the basolateral amygdala, the bed nucleus of the stria terminalis [BNST] and the prefrontal cortex). Methods: We monitored the activation pattern of these regions (measured by Fos) during cocaine-seeking induced by OrxA administered to the pPVT. The BNST and CeA emerged as being selectively activated. To test whether the functionality of these regions was pivotal during OrxA-induced cocaine-seeking behaviour, we transiently inactivated these regions concomitantly with OrxA administration to the pPVT. We then tested the participation of corticotropin-releasing factor receptors (CRF1) in the CeA during OrxA-induced cocaine-seeking using the CRF1 antagonist CP154526. Results: We observed selective activation of the CeA and BNST during cocaine-seeking induced by OrxA administered to the pPVT, but only transient inactivation of the CeA prevented cocaine-seeking behaviour. Administration of CP154526 to the CeA prevented OrxAinduced cocaine-seeking behaviour. Limitations: The use of only male rats could have been a limitation. Other limitations could have been the use of an indirect approach to test the hypothesis that administration of OrxA to the pPVT drives cocaine-seeking via CRF1 signalling in the CeA, and a lack of analysis of the participation of CeA subregions. Conclusion: Cocaine-seeking behaviour induced by OrxA administered to the pPVT is driven by activation of the CeA via CRF1 signalling.
Asunto(s)
Núcleo Amigdalino Central/efectos de los fármacos , Trastornos Relacionados con Cocaína/prevención & control , Cocaína , Orexinas/farmacología , Núcleo Hipotalámico Paraventricular/efectos de los fármacos , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Tálamo/efectos de los fármacos , Animales , Cocaína/farmacología , Masculino , Orexinas/administración & dosificación , RatasRESUMEN
Hyperactivity of the hypothalamic-pituitary-adrenal axis and impairment of the central corticotropin-releasing factor system are factors in the pathogenesis of depression. Though several antagonists of the corticotropin-releasing factor 1 receptor were effective in the recognized behavioral tests for antidepressant activity, there is still little information on the potential interactions between corticotropin-releasing factor 1 receptor inhibitors and conventional antidepressant therapy. The aim of our study was to assess the influence of CP154526, a corticotropin-releasing factor 1 receptor blocker, which presented some signs of depression. Our results revealed that CP154526 (5 and 10 mg/kg) or fluoxetine (10 mg/kg) treatment notably improved the sucrose consumption, produced anti-depressive-like behavior in open-field test, as well as immobility time in forced swimming test. The levels of interleukin-6, interleukin-1ß, tumor necrosis factor-α, and corticotropin-releasing hormone concentration in the serum were inhibited effectively by CP154526 or fluoxetine administration. Real-time quantitative PCR and western blot analysis showed the upregulated levels of brain-derived neurotrophic factor and growth associated protein 43 (GAP43) in the hypothalamus of the rats exposed to chronic unpredictable mild stress (CUMS), while different degrees of downregulation in their expression were detected after CP154526 (5 and 10 mg/kg) or fluoxetine (10 mg/kg) treatment, respectively. Thus, our data demonstrated that CP154526 exhibited antidepressant effect in CUMS rats, which might be mediated by decreasing the brain-derived neurotrophic factor and GAP43 expression in the hypothalamus.
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Antidepresivos/farmacología , Depresión/etiología , Depresión/metabolismo , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Estrés Psicológico/complicaciones , Animales , Conducta Animal/efectos de los fármacos , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Proteína GAP-43/metabolismo , Hipotálamo/efectos de los fármacos , Hipotálamo/metabolismo , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
Overactive bladder (OAB) often co-exists with depression in women. The corticotropin-releasing factor (CRF) system participates in the pathophysiology of both disorders. Therefore, we tested the effects of acute treatment with a reversible CRF receptor type-1 (CRF1) antagonist, SN003 (1mg/kg, i.v.), representatives of first (solifenacin, 0.03mg/kg, i.v.) and second (mirabegron, 1mg/kg, i.v.) line treatments for OAB as well as an antidepressant imipramine (30mg/kg, i.p.) on changes in depressive-like behavior and detrusor overactivity (DO) symptoms induced by a 6-week administration of 13-cis-retinoic acid (13-cis-RA, 1mg/kg/day, i.p.) in female Wistar rats, using in vivo cystometric investigations, forced swim test (FST) and spontaneous locomotor activity test. Following cystometric and behavioral studies, tissue was harvested and CRF level was assessed in the hypothalamus, amygdala and plasma. 13-cis-RA-induced depressive-like behavior and DO symptoms were associated with increased CRF levels in the hypothalamus, amygdala and plasma. Solifenacin and mirabegron attenuated DO symptoms induced by 13-cis-RA, did not display antidepressant-like activity and did not influence CRF levels in brain tissues or plasma. Imipramine and SN003 displayed antidepressant-like activity and lowered increased levels of CRF in brain tissues and plasma. Imipramine attenuated changes in some of the cystometric parameters, which are associated with OAB dry (without urge incontinence), whereas SN003 attenuated changes in almost all cystometric parameters that were induced by 13-cis-RA. CRF1 antagonist may be beneficial in case of OAB wet (with urge incontinence) or dry co-existing with depression. The possible mechanism may be related to the effects on central/peripheral CRF system.
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Conducta Animal/efectos de los fármacos , Depresión/tratamiento farmacológico , Isotretinoína/farmacología , Piridinas/farmacología , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Triazoles/farmacología , Vejiga Urinaria Hiperactiva/inducido químicamente , Vejiga Urinaria Hiperactiva/tratamiento farmacológico , Amígdala del Cerebelo/efectos de los fármacos , Amígdala del Cerebelo/metabolismo , Animales , Hormona Liberadora de Corticotropina/sangre , Hormona Liberadora de Corticotropina/metabolismo , Depresión/complicaciones , Depresión/metabolismo , Relación Dosis-Respuesta a Droga , Femenino , Hipotálamo/efectos de los fármacos , Hipotálamo/metabolismo , Piridinas/uso terapéutico , Ratas , Ratas Wistar , Triazoles/uso terapéutico , Vejiga Urinaria Hiperactiva/complicaciones , Vejiga Urinaria Hiperactiva/metabolismoRESUMEN
Hyperactivity of the hypothalamic-pituitary-adrenal axis (HPA) and impairment of the central corticotropin-releasing factor (CRF) system are factors in the pathogenesis of depression. Though several antagonists of the CRF1 receptor were effective in the recognized behavioral tests for antidepressant activity, there is still little information on the potential interactions between CRF1 receptor inhibitors and conventional antidepressant therapy. The aim of our study was to assess the influence of SN003, a CRF1 receptor blocker, on the activity of imipramine and fluoxetine in the forced swim test (FST) in rats which presented some signs of depression. The experiments were carried out on female Wistar rats subjected to 14-day subcutaneous corticosterone (CORT) administration (20 mg/kg/day). The antidepressant-like effect was determined by the FST and the CRF levels in the hypothalamus, amygdala, and peripheral blood were measured by a high-sensitivity immunoenzymatic test. SN003 (0.5 mg/kg) potentiated the antidepressant-like effect of imipramine (15 mg/kg) and fluoxetine (7.5 mg/kg). Moreover, the co-administration of the tested agents abolished CORT-induced increase in CRF levels in the examined biological material more profoundly than monotherapy. Our present findings give further evidence that the blockage of CRF action may be useful in the treatment of mood disorders. The concurrent use of well-known antidepressants with CRF1 receptor antagonists could be beneficial in terms of safety, since it requires lower doses of the applied agents.
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Antidepresivos/uso terapéutico , Corticosterona/farmacología , Depresión/tratamiento farmacológico , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Amígdala del Cerebelo/efectos de los fármacos , Amígdala del Cerebelo/metabolismo , Animales , Antidepresivos/farmacología , Conducta Animal/efectos de los fármacos , Hormona Liberadora de Corticotropina/sangre , Hormona Liberadora de Corticotropina/metabolismo , Depresión/metabolismo , Quimioterapia Combinada , Femenino , Fluoxetina/farmacología , Fluoxetina/uso terapéutico , Hipotálamo/efectos de los fármacos , Hipotálamo/metabolismo , Imipramina/farmacología , Imipramina/uso terapéutico , Locomoción/efectos de los fármacos , Piridinas/farmacología , Piridinas/uso terapéutico , Ratas Wistar , Receptores de Hormona Liberadora de Corticotropina/metabolismo , Natación , Triazoles/farmacología , Triazoles/uso terapéuticoRESUMEN
Rikkunshito (RKT), a Kampo medicine, has been reported to show an ameliorative effect on sustained hypophagia after novelty stress exposure in aged mice through serotonin 2C receptor (5-HT2CR) antagonism. We aimed to determine (1) whether the activation of anorexigenic neurons, corticotropin-releasing factor (CRF), and pro-opiomelanocortin (POMC) neurons, is involved in the initiation of hypophagia induced by novelty stress in aged mice; (2) whether the ameliorative effect of RKT is associated with CRF and POMC neurons and downstream signal transduction; and (3) the plasma and brain distribution of the active components of RKT. The administration of RKT or 5-HT2CR, CRF receptor 1 (CRFR1), and melanocortin-4 receptor antagonists significantly restored the decreased food intake observed in aged male C57BL/6 mice in the early stage after novelty stress exposure. Seven components of RKT exhibited antagonistic activity against CRFR1. Hesperetin and isoliquiritigenin, which showed antagonistic effects against both CRFR1 and 5-HT2CR, were distributed in the plasma and brain of male Sprague-Dawley rats after a single oral administration of RKT. In conclusion, the ameliorative effect of RKT in this model is assumed to be at least partly due to brain-distributed active components possessing 5-HT2CR and CRFR1 antagonistic activities.
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Anorexia/prevención & control , Encéfalo/metabolismo , Medicamentos Herbarios Chinos/farmacología , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Estrés Psicológico/complicaciones , Animales , Anorexia/etiología , Masculino , Ratones , Ratas , Ratas Sprague-Dawley , Receptores de Hormona Liberadora de Corticotropina/metabolismoRESUMEN
Blockade of corticotropin-releasing factor receptor 1 (CRF1) suppresses stress-induced alcohol seeking in rodents, but clinical translation remains. Here, we first showed that the CRF1 antagonist verucerfont potently blocks hypothalamic-pituitary adrenal (HPA) axis activation in adrenalectomized rats. We then evaluated verucerfont for its ability to block HPA axis activation and reduce stress-induced alcohol craving in alcohol-dependent patients. Anxious, alcohol-dependent women (age 21-65 years, n=39) were admitted to the NIH Clinical Center and completed withdrawal treatment before enrollment if needed. One-week single-blind placebo was followed by randomized double-blind verucerfont (350 mg per day) or placebo for 3 weeks. Verucerfont effects on the HPA axis were evaluated using the dexamethasone-CRF test. Craving was evaluated using two established protocols, one that combines a social stressor with physical alcohol cue exposure, and one that uses guided imagery to present personalized stress, alcohol, or neutral stimuli. An fMRI session examined brain responses to negative affective stimuli and alcohol cues. In contrast to our recent observations with another CRF1 antagonist, pexacerfont, verucerfont potently blocked the HPA axis response to the dexamethasone-CRF test, but left alcohol craving unaffected. Right amygdala responses to negative affective stimuli were significantly attenuated by verucerfont, but responses to alcohol-associated stimuli were increased in some brain regions, including left insula. Discontinuation rates were significantly higher in the verucerfont group. Our findings provide the first translational evidence that CRF1 antagonists with slow receptor dissociation kinetics may have increased efficacy to dampen HPA axis responses. The findings do not support a clinical efficacy of CRF1 blockade in stress-induced alcohol craving and relapse.
Asunto(s)
Hormona Adrenocorticotrópica/sangre , Alcoholismo/tratamiento farmacológico , Ansiedad/tratamiento farmacológico , Compuestos de Azabiciclo/uso terapéutico , Hidrocortisona/sangre , Oxadiazoles/uso terapéutico , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Adrenalectomía , Adulto , Anciano , Alcoholismo/complicaciones , Alcoholismo/diagnóstico por imagen , Animales , Ansiedad/diagnóstico por imagen , Ansiedad/etiología , Ansia/efectos de los fármacos , Modelos Animales de Enfermedad , Método Doble Ciego , Femenino , Humanos , Procesamiento de Imagen Asistido por Computador , Imágenes en Psicoterapia , Persona de Mediana Edad , Oxígeno/sangre , Escalas de Valoración Psiquiátrica , Ratas , Ratas Sprague-Dawley , Estudios Retrospectivos , Método Simple Ciego , Adulto JovenRESUMEN
Cocaine-and amphetamine-regulated transcript peptides (CARTp) suppress gastric emptying and nutritional intake following 4th icv administration. Whereas, the CARTp inhibition of gastric emptying was blocked by pre-treatment with a non-selective corticotropin releasing factor (CRF) antagonist, sucrose drinking was not, suggesting that CARTp- and CRF controls for food intake and gastric emptying are operated through separable dorsal hindbrain mechanisms. The aim of the study was to explore CARTp-CRF brainstem interactions on patterns of neuronal activation in areas of the brainstem and midbrain relevant to gastrointestinal control and feeding regulation. Rats received 4th icv injections of combinations of vehicle, CARTp (1µg), or the nonselective CRF antagonist, α-helical CRF9-41 (αCRF), in a randomized order. Brain sections were processed for c-fos by immunohistochemistry followed by image analysis at defined levels of the brain. CARTp (1µg, 4th icv) induced a robust c-fos response in the nucleus of the solitary tract (NTS) and area postrema (AP), whereas, no c-fos could be detected in the parabrachial nucleus (PBN), the paraventricular nucleus of the hypothalamus (PVN) or the arcuate nucleus of the hypothalamus (ARC). The c-fos expression in the structures of the dorsal vagal complex (DVC) was completely blocked by pre-treatment with the CRF antagonist, which did not by itself induce c-fos at any examined level. After CARTp and αCRF in combination, there was a tendency towards an increased c-fos response in the ARC. We conclude that CARTp activates cells of the area postrema and NTS via a downstream, CRF-dependent mechanism.
Asunto(s)
Área Postrema/metabolismo , Proteínas del Tejido Nervioso/farmacología , Proteínas Proto-Oncogénicas c-fos/biosíntesis , Núcleo Solitario/metabolismo , Animales , Hormona Liberadora de Corticotropina/farmacología , Ingestión de Alimentos/fisiología , Cuarto Ventrículo , Tracto Gastrointestinal/fisiología , Hipotálamo/efectos de los fármacos , Hipotálamo/metabolismo , Masculino , Mesencéfalo/efectos de los fármacos , Mesencéfalo/metabolismo , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Fragmentos de Péptidos/farmacología , Ratas Sprague-Dawley , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Receptores de Hormona Liberadora de Corticotropina/metabolismoRESUMEN
The primary cause of the neurodegenerative process that underlies Parkinson's disease (PD) is still unknown. Different mechanisms probably contribute to triggering neuronal death in the nigro-striatum pathway. The neuropeptide urocortin 2 (UCN2) plays an important role in the regulation of striatum (STR) neurons projection. We investigated the effects of UCN2 on spontaneous discharge and glutamatergic responses in STR for a better understanding of the pathogenesis of PD. The experiment used microiontophoresis method to observe the effects of UCN2 on STR neurons' firing rates in vivo. Corticotrophin releasing factor receptor 2 (CRF-R2) selective inhibitor, astressin-2B (AST-2B), was administered simultaneously with UCN2 to investigate the effects of UCN2 on CRF-R2. Moreover, we further explored the effects of UCN2 on glutamatergic responses in STR neurons. We found that UCN2 could significantly inhibit the firing rate of 84% of the tested STR neurons, and its inhibitory effect followed a concentration-dependent manner. During the microiontophoresis of GLU, the excitatory firing of glutamatergic neurons could be attenuated by the addition of UCN2, but enhanced by the application of AST-2B. The results suggest that UCN2 could regulate the effects of STR neurotransmitters (GLU) via CRF-R2 and may thereby contribute to the improvement of PD.
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Cuerpo Estriado/citología , Hormona Liberadora de Corticotropina/farmacología , Ácido Glutámico/fisiología , Neuronas/efectos de los fármacos , Transmisión Sináptica/efectos de los fármacos , Urocortinas/farmacología , Potenciales de Acción/efectos de los fármacos , Animales , Antiparkinsonianos/farmacología , Cuerpo Estriado/efectos de los fármacos , Depresión Química , Maleato de Dizocilpina/farmacología , Relación Dosis-Respuesta a Droga , Evaluación Preclínica de Medicamentos , Antagonistas de Aminoácidos Excitadores/farmacología , Iontoforesis , Masculino , Neuronas/fisiología , Fragmentos de Péptidos/farmacología , Péptidos Cíclicos/farmacología , Ratas , Ratas Sprague-Dawley , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidoresRESUMEN
Fear to predictable threat and anxiety to unpredictable threat reflect distinct processes mediated by different brain structures, the central nucleus of the amygdala and the bed nucleus of the stria terminalis (BNST), respectively. This study tested the hypothesis that the corticotropin-releasing factor (CRF1) antagonist GSK561679 differentially reduces anxiety but increases fear in humans. A total of 31 healthy females received each of four treatments: placebo, 50 mg GSK561679 (low-GSK), 400 mg GSK561679 (high-GSK), and 1 mg alprazolam in a crossover design. Participants were exposed to three conditions during each of the four treatments. The three conditions included one in which predictable aversive shocks were signaled by a cue, a second during which shocks were administered unpredictably, and a third condition without shock. Fear and anxiety were assessed using the acoustic startle reflex. High-GSK had no effect on startle potentiation during unpredictable threat (anxiety) but increased startle potentiation during the predictable condition (fear). Low-GSK did not affect startle potentiation across conditions. Consistent with previous findings, alprazolam reduced startle potentiation during unpredictable threat but not during predictable threat. The increased fear by high-GSK replicates animal findings and suggests a lift of the inhibitory effect of the BNST on the amygdala by the CRF1 antagonist.
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Ansiedad/inducido químicamente , Compuestos de Azabiciclo/farmacología , Miedo/efectos de los fármacos , Oxadiazoles/farmacología , Psicotrópicos/farmacología , Reflejo de Sobresalto/efectos de los fármacos , Estimulación Acústica , Adulto , Alprazolam/farmacología , Ansiolíticos/farmacología , Anticipación Psicológica/fisiología , Ansiedad/fisiopatología , Compuestos de Azabiciclo/efectos adversos , Estudios Cruzados , Relación Dosis-Respuesta a Droga , Método Doble Ciego , Electrochoque , Miedo/fisiología , Femenino , Humanos , Pruebas Neuropsicológicas , Oxadiazoles/efectos adversos , Psicotrópicos/efectos adversos , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Receptores de Hormona Liberadora de Corticotropina/metabolismo , Reflejo de Sobresalto/fisiología , Encuestas y Cuestionarios , IncertidumbreRESUMEN
Corticotropin-releasing factor (CRF) plays a critical role in the mediation of physiological and behavioral responses to stressors. In the present study, we investigated the role played by the CRF system within the dorsomedial hypothalamus (DMH) in the modulation of anxiety- and panic-related responses. Male Wistar rats were administered into the DMH with CRF (125 and 250 ng/0.2 µl, experiment 1) or with the CRFR1 antagonist antalarmin (25 ng/0.2 µl, experiment 2) and 10 min later tested in the elevated T-maze (ETM) for inhibitory avoidance and escape measurements. In clinical terms, these responses have been respectively related to generalized anxiety and panic disorder. To further verify if the anxiogenic effects of CRF were mediated by CRFR1 activation, we also investigated the effects of the combined treatment with CRF (250 ng/0.2 µl) and antalarmin (25 ng/0.2 µl) (experiment 3). All animals were tested in an open field, immediately after the ETM, for locomotor activity assessment. Results showed that 250 ng/0.2µl of CRF facilitated ETM avoidance, an anxiogenic response. Antalarmin significantly decreased avoidance latencies, an anxiolytic effect, and was able to counteract the anxiogenic effects of CRF. None of the compounds administered altered escape responses or locomotor activity measurements. These results suggest that CRF in the DMH exerts anxiogenic effects by activating type 1 receptors, which might be of relevance to the physiopathology of generalized anxiety disorder.
Asunto(s)
Reacción de Prevención/efectos de los fármacos , Hormona Liberadora de Corticotropina/farmacología , Hipotálamo/efectos de los fármacos , Aprendizaje por Laberinto/efectos de los fármacos , Pirimidinas/farmacología , Pirroles/farmacología , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Animales , Ansiedad/tratamiento farmacológico , Reacción de Fuga/efectos de los fármacos , Masculino , Actividad Motora/efectos de los fármacos , Ratas , Ratas WistarRESUMEN
Adrenomedullin (AM), a 52 residue neuropeptide, is associated with anorexia in mammals and has a poorly understood central mechanism of action. Thus, this study focused on elucidating AM's central mechanism of action in an alternative vertebrate model, the chick (Gallus gallus). In Experiment 1, chicks centrally injected with AM dose-dependently reduced food but not water intake. In Experiment 2, those chicks that received central AM had increased c-Fos immunoreactivity in the magnocellular division of the paraventricular nucleus (PaMC), ventromedial hypothalamus (VMH) and doromedial hypothalamus (DM). The lateral hypothalamic area, parvocellular division of the paraventricular hypothalamus and the arcuate nucleus were not affected. In Experiment 3, antagonism of corticotrophin releasing factor (CRF) receptors did not affect AM-associated anorexia. In Experiment 4, a comprehensive behavior analysis was conducted and AM-treated chicks pecked less, moved more, jumped more and spent more time in deep rest. In conclusion, exogenous AM induced anorexia is associated with activation of the PaMC, VMH and DM of the hypothalamus, is not CRF dependent, and affects behaviors unrelated to food intake in chicks.
Asunto(s)
Adrenomedulina/farmacología , Anorexia , Antihipertensivos/farmacología , Pollos/metabolismo , Hipotálamo/efectos de los fármacos , Animales , Anorexia/metabolismo , Conducta Animal/efectos de los fármacos , Pollos/crecimiento & desarrollo , Ingestión de Líquidos/efectos de los fármacos , Ingestión de Alimentos/efectos de los fármacos , Hipotálamo/metabolismo , Inyecciones Intraventriculares , Proteínas Proto-Oncogénicas c-fos/metabolismo , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Receptores de Hormona Liberadora de Corticotropina/metabolismoRESUMEN
RATIONALE: As enhanced corticotropin-releasing factor (CRF) transmission is associated with induction of sensorimotor gating deficits, CRF1 receptor antagonists may reverse disrupted prepulse inhibition (PPI), an operational measure of sensorimotor gating. OBJECTIVES: To determine the effects of CRF1 receptor antagonists in pharmacological models of disrupted PPI and to determine if long-term elevated central CRF levels alter sensitivity towards PPI disrupting drugs. METHODS: CP154,526 (10-40 mg/kg), SSR125543 (3-30 mg/kg) and DMP695 (40 mg/kg) were tested on PPI disruption provoked by D-amphetamine (2.5, 3 mg/kg), ketamine (5, 30 mg/kg) and MK801 (0.2, 0.5 mg/kg) in Wistar rats, C57Bl/6J and CD1 mice, and on spontaneously low PPI in Iffa Credo rats and DBA/2J mice. PPI-disrupting effects of D-amphetamine (2.5-5 mg/kg) and MK801 (0.3-1 mg/kg) were examined in CRF-overexpressing (CRFtg) mice, which display PPI deficits. Finally, we determined the influence of CP154,526 on D-amphetamine-induced dopamine outflow in nucleus accumbens and prefrontal cortex of CRFtg mice using in vivo microdialysis. RESULTS: No CRF1-antagonists improved PPI deficits in any test. CRFtg mice showed blunted PPI disruption in response to MK801, but not D-amphetamine. Further, D-amphetamine-induced dopamine release was less pronounced in CRFtg versus wild-type mice, a response normalized by pretreatment with CP154,526. CONCLUSION: The inability of CRF1 receptor antagonists to block pharmacological disruption of sensorimotor gating suggests that the involvement of CRF1 receptors in the modulation of dopaminergic and glutamatergic neurotransmission relevant for sensory gating is limited. Furthermore, the alterations observed in CRFtg mice support the notion that long-term elevated central CRF levels induce changes in these neurotransmitter systems.
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Hidrocarburos Halogenados/farmacología , Inhibición Prepulso/efectos de los fármacos , Piridinas/farmacología , Pirimidinas/farmacología , Pirroles/farmacología , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Tiazinas/farmacología , Estimulación Acústica , Animales , Hormona Liberadora de Corticotropina/genética , Dextroanfetamina/antagonistas & inhibidores , Dextroanfetamina/farmacología , Maleato de Dizocilpina/antagonistas & inhibidores , Maleato de Dizocilpina/farmacología , Dopamina/metabolismo , Relación Dosis-Respuesta a Droga , Ketamina/antagonistas & inhibidores , Ketamina/farmacología , Masculino , Ratones , Ratones Transgénicos , Núcleo Accumbens/efectos de los fármacos , Núcleo Accumbens/metabolismo , Corteza Prefrontal/efectos de los fármacos , Corteza Prefrontal/metabolismo , Ratas , Reflejo de Sobresalto/efectos de los fármacosRESUMEN
Corticotropin-releasing factor (CRF) and corticotropin-releasing factor receptor (CRFR) play important roles in stress response, including anxiety and depression syndrome. The CRF expression also relates to chronic stress-related hair loss. This study utilizes the world's largest traditional Chinese medicine (TCM) database and molecular dynamics (MD) simulations to investigate novel CRFR inhibitors for treatment of alopecia. The docking and screening from TCM database results indicate the vitamin B2, 3 beta-isodihydrocadambine, and caribine display higher binding affinity than maltose in maltose binding protein (MBP). However, the results of MD simulation shows the caribine-facilitated CRFR approach closer to MBP, the 3D structure conformation of MBP and CRFR complex forms compact structure. Interestingly, the distance between the two proteins is reducing significantly after caribine dock into MBP binding site. Beside, from Ligand channel analysis, the paths of caribine demonstrate that residence time is increased in binding pocket. Hence, our finding suggests that caribine might be a potential lead compound to stimulate MBP and CRFR interaction, and help for baldless therapy in further study.
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Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/uso terapéutico , Enfermedades del Cabello/tratamiento farmacológico , Medicina Tradicional China , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Análisis por Conglomerados , Medicamentos Herbarios Chinos/química , Humanos , Ligandos , Simulación del Acoplamiento Molecular , Análisis de Componente Principal , Receptores de Hormona Liberadora de Corticotropina/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , TermodinámicaRESUMEN
Exposure to a stressor sensitizes behavioral and hormonal responses to future stressors. Stress-associated release of noradrenaline enhances the capacity of central synapses to show plasticity (metaplasticity). We found noradrenaline-dependent metaplasticity at GABA synapses in the paraventricular nucleus of the hypothalamus in rat and mouse that controls the hypothalamic-pituitary-adrenal axis. In vivo stress exposure was required for these synapses to undergo activity-dependent long-term potentiation (LTPGABA). The activation of ß-adrenergic receptors during stress functionally upregulated metabotropic glutamate receptor 1 (mGluR1), allowing for mGluR1-dependent LTPGABA during afferent bursts. LTPGABA was expressed postsynaptically and manifested as the emergence of new functional synapses. Our findings provide, to the best of our knowledge, the first demonstration that noradrenaline release during an in vivo challenge alters information storage capacity at GABA synapses. Because these GABA synapses become excitatory following acute stress, this metaplasticity may contribute to neuroendocrine sensitization to stress.
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Plasticidad Neuronal/fisiología , Norepinefrina/metabolismo , Estrés Psicológico/metabolismo , Sinapsis/metabolismo , Ácido gamma-Aminobutírico/metabolismo , Animales , Animales Recién Nacidos , Channelrhodopsins , Quelantes/farmacología , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Modelos Animales de Enfermedad , Inhibidores Enzimáticos/farmacología , Hipotálamo/citología , Técnicas In Vitro , Potenciales Postsinápticos Inhibidores/efectos de los fármacos , Potenciales Postsinápticos Inhibidores/genética , Luz , Proteínas Luminiscentes/genética , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Plasticidad Neuronal/efectos de los fármacos , Neurotransmisores/farmacología , Ratas , Ratas Sprague-Dawley , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Receptores de Glutamato Metabotrópico/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Estrés Psicológico/inducido químicamente , Estrés Psicológico/patología , Sinapsis/efectos de los fármacosRESUMEN
Neuropeptide W (NPW) was isolated as an endogenous ligand for NPBWR1, an orphan G protein-coupled receptor localized in the rat brain, including the paraventricular nucleus. It has been reported that central administration of NPW stimulates corticosterone secretion in rats. We hypothesized that NPW activates the hypothalamic-pituitary-adrenal (HPA) axis via corticotrophin-releasing factor (CRF) and/or arginine vasopressin (AVP). NPW at 1 pM to 10 nM did not affect basal or ACTH-induced corticosterone release from dispersed rat adrenocortical cells, or basal and CRF-induced ACTH release from dispersed rat anterior pituitary cells. In conscious and unrestrained male rats, intravenous administration of 2.5 and 25 nmol NPW did not affect plasma ACTH levels. However, intracerebroventricular (icv) administration of 2.5 and 5.0 nmol NPW increased plasma ACTH levels in a dose-dependent manner at 15 min after stimulation (5.0 vs. 2.5 nmol NPW vs. vehicle: 1802 ± 349 vs. 1170 ± 204 vs. 151 ± 28 pg/mL, respectively, mean ± SEM). Pretreatment with astressin, a CRF receptor antagonist, inhibited the increase in plasma ACTH levels induced by icv administration of 2.5 nmol NPW at 15 min (453 ± 176 vs. 1532 ± 343 pg/mL, p<0.05) and at 30 min (564 ± 147 vs. 1214 ± 139 pg/mL, p<0.05) versus pretreatment with vehicle alone. However, pretreatment with [1-(ß-mercapto-ß, ß-cyclopentamethylenepropionic acid), 2-(Ο-methyl)tyrosine]-arg-vasopressin, a V1a/V1b receptor antagonist, did not affect icv NPW-induced ACTH release at any time (p>0.05). In conclusion, we suggest that central NPW activates the HPA axis by activating hypothalamic CRF but not AVP.
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Hormona Adrenocorticotrópica/metabolismo , Arginina Vasopresina/fisiología , Hormona Liberadora de Corticotropina/fisiología , Neuropéptidos/farmacología , Hormona Adrenocorticotrópica/sangre , Animales , Antagonistas de los Receptores de Hormonas Antidiuréticas , Células Cultivadas , Corticosterona/metabolismo , Evaluación Preclínica de Medicamentos , Antagonistas de Hormonas/administración & dosificación , Antagonistas de Hormonas/farmacología , Inyecciones Intravenosas , Masculino , Neuropéptidos/administración & dosificación , Adenohipófisis/citología , Adenohipófisis/efectos de los fármacos , Adenohipófisis/metabolismo , Ratas , Ratas Sprague-Dawley , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Regulación hacia Arriba/efectos de los fármacosRESUMEN
Although acute stress accelerates colonic transit, the effect of chronic stress on colonic transit remains unclear. In this study, rats received repeated restraint stress (chronic homotypic stress) or various types of stress (chronic heterotypic stress) for 5 and 7 days, respectively. Vehicle saline, oxytocin (OXT), OXT receptor antagonist or corticotropin-releasing factor (CRF) receptor antagonists were administered by intracerebroventricular (ICV) injection prior to restraint stress for 90 min. Immediately after the stress exposure, the entire colon was removed and the geometric center (GC) of Na51CrO4 (a nonabsorbable radioactive marker; 0.5 µCi) distribution was calculated to measure the transit. Gene expression of OXT and CRF in the paraventricular nucleus (PVN) was evaluated by in situ hybridization. Accelerated colonic transit with the acute stressor was no longer observed following chronic homotypic stress. This restored colonic transit was reversed by ICV injection of an OXT antagonist. In contrast, chronic heterotypic stress significantly accelerated colonic transit, which was attenuated by ICV injection of OXT and by a CRF receptor 1 antagonist. OXT mRNA expression in the PVN was significantly increased following chronic homotypic stress, but not chronic heterotypic stress. However, CRF mRNA expression in the PVN was significantly increased following acute and chronic heterotypic stress, but not chronic homotypic stress. These results indicate that central OXT and CRF play a pivotal role in mediating the colonic dysmotility following chronic stress in rats.
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Colon/metabolismo , Hormona Liberadora de Corticotropina/metabolismo , Hipotálamo/metabolismo , Oxitocina/metabolismo , Estrés Fisiológico/fisiología , Animales , Colon/efectos de los fármacos , Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Hormona Liberadora de Corticotropina/genética , Expresión Génica/fisiología , Masculino , Oxitocina/genética , Oxitocina/farmacología , Núcleo Hipotalámico Paraventricular/metabolismo , Ratas , Ratas Sprague-Dawley , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Receptores de Oxitocina/antagonistas & inhibidores , Receptores de Oxitocina/metabolismo , Restricción Física/fisiología , Estrés PsicológicoRESUMEN
In this study, we tested the effect of urocortin 1 (Ucn1) on the contractility of gallbladder smooth muscle (GBSM) strips from guinea pigs and studied the involvement of corticotropin-releasing factor (CRF) receptors in this effect. The effect of Ucn1 on the isometric contractions of non-contracted and acetylcholine (Ach)-contracted GBSM, and the effects of CRF-R antagonists antalarmin and astressin 2B on the effect of Ucn1 were studied. In addition, the expression of receptors for CRF-R1 and CRF-R2 in guinea pig gallbladder were investigated using reverse transcription - polymerase chain reaction (RT-PCR). Ucn1 dose-dependently inhibited the contractility of GBSM. Moreover, Ucn1 decreased the resting tension, the mean contractile amplitude, and the contractile frequency in both non-contracted and Ach-contracted strips of GBSM. Furthermore, Ucn1 induced rightward shift of the Ach concentration-response curve of Ach in Ach-contracted strips. This inhibitory effect of Ucn1 on both non-contracted and Ach-contracted strips was inhibited by astressin 2B, but not by antalarmin. RT-PCR demonstrated that the CRF-R2, but not CRF-R1 receptor subtype is expressed in the muscularis muscle of guinea pig gallbladder. In conclusion, Ucn1 has an inhibitory effect on the contractility of GBSM of guinea pig, mediated through stimulating CRF-R2 receptors in GBSM. More studies are needed to clarify the intracellular signaling events involved in this effect.
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Vesícula Biliar/efectos de los fármacos , Contracción Muscular/efectos de los fármacos , Músculo Liso/efectos de los fármacos , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Urocortinas/fisiología , Acetilcolina/metabolismo , Animales , ADN Complementario/análisis , Relación Dosis-Respuesta a Droga , Evaluación Preclínica de Medicamentos , Vesícula Biliar/metabolismo , Vesícula Biliar/fisiología , Cobayas , Técnicas In Vitro , Masculino , Contracción Muscular/fisiología , Músculo Liso/metabolismo , Músculos/metabolismo , Fragmentos de Péptidos/farmacología , Péptidos Cíclicos/farmacología , Pirimidinas/farmacología , Pirroles/farmacología , Receptores de Hormona Liberadora de Corticotropina/metabolismo , Transducción de Señal/efectos de los fármacos , Urocortinas/análisisRESUMEN
Despite the availability of X-ray crystal structure data for several members of the G-protein-coupled receptor (GPCR) superfamily, structure-based discovery of GPCR ligands has been exclusively restricted to class A (rhodopsin-like) receptors. Herein we report the identification, by a docking-based virtual screening approach, of noncompetitive ligands for two related class B (secretin-like) GPCRs: the glucagon receptor (GLR) and the glucagon-like peptide 1 receptor (GLP-1R). Starting from a knowledge-based three-dimensional model of the GLR, a database of 1.9 million commercially available drug-like compounds was screened for chemical similarity to existing GLR noncompetitive antagonists and docked to the transmembrane cavity of the GLR; 23 compounds were then selected based on protein-ligand interaction fingerprints, and were then purchased and evaluated for in vitro binding to GLR and modulation of glucagon-induced cAMP release. Two of the 23 compounds inhibited the effect of glucagon in a dose-dependent manner, with one inhibitor exhibiting the same potency as L-168 049, a reference noncompetitive GLR antagonist, in a whole-cell-based functional assay. Interestingly, one virtual hit that was inactive at the GLR was shown to bind to GLP-1R and potentiate the response to the endogenous GLP-1 ligand.
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Receptores de Glucagón/antagonistas & inhibidores , Regulación Alostérica , Secuencia de Aminoácidos , Sitios de Unión , Bases de Datos Factuales , Evaluación Preclínica de Medicamentos , Humanos , Ligandos , Simulación de Dinámica Molecular , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Estructura Terciaria de Proteína , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Receptores de Hormona Liberadora de Corticotropina/metabolismo , Receptores de Glucagón/genética , Receptores de Glucagón/metabolismo , Alineación de Secuencia , Relación Estructura-ActividadRESUMEN
Bipolar electrodes were implanted in the lateral hypothalamus in a group of 44 Wistar male rats in order to study self-stimulation reaction in the Skinner box. Simultaneously, microcanules were implanted into the central nucleus of the amygdala to inject the drugs (1 microl per injection). The blockade of corticoliberin (CRF) receptors (astressin, 1 microg) or Na+influx currents (xycaine or lidocain 1 microg) by the intrastructural administration of drugs into the amygdala decreased self-stimulation reaction of the lateral hypothalamus in rats by 29-55%. The inhibition of D1 and D2 dopamine receptors in the amygdala with SCH23390 (1 microg) or sulpiride (1 microg) respectively, also reduced self-stimulation but to a lower degree. On the background of blockade of CRF (astressin) and dopamine (sulpiride) receptors as well as sodium influx ionic currents (lidocain) in the amygdala neurons, psychomotor stimulant amphetamine (1 mg/kg) and barbiturate sodium ethaminal (5 mg/kg) retained their psychoactivating effect on self-stimulation (+30-37%), while fentanyl (0.1 mg/kg) and leu-enkephaline (0.1 mg/kg) did not produce this effect. Fentanyl moderately activated self-stimulation only after the blockade of D1 dopamine receptors with SCH23390. After the blockade of CRF receptors, leu-enkephaline strengthened its depressant effect on self-stimulation reaction (-89%). Therefore, if the modulating action of amygdala on the hypothalamus is eliminated, the enhancing effects of opiates (fentanyl) and opioids (leu-encephaline) are blocked, but the effects of psychomotor stimulant amphetamine and barbiturate sodium ethaminal are retained.
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Amígdala del Cerebelo/efectos de los fármacos , Hipotálamo/efectos de los fármacos , Receptores de Hormona Liberadora de Corticotropina/metabolismo , Receptores Dopaminérgicos/metabolismo , Autoestimulación/fisiología , Anfetamina/farmacología , Amígdala del Cerebelo/fisiología , Analgésicos Opioides/farmacología , Animales , Benzazepinas/farmacología , Cateterismo , Hormona Liberadora de Corticotropina/farmacología , Dopamina/farmacología , Antagonistas de Dopamina/farmacología , Electrodos Implantados , Fentanilo/farmacología , Hipotálamo/fisiología , Masculino , Narcóticos/farmacología , Fragmentos de Péptidos/farmacología , Ratas , Ratas Wistar , Receptores de Hormona Liberadora de Corticotropina/antagonistas & inhibidores , Sulpirida/farmacologíaRESUMEN
Corticotropin-releasing factor (CRF) and CRF-related peptides exert hypophysiotropic and anorexigenic effects in mammals and teleost fish. In anuran amphibians, CRF acts as a potent stimulator of thyrotropin release from the pituitary. According to our recent study, CRF also acts as an anorexigenic factor for the cessation of food intake in the metamorphosing bullfrog larvae. However, the anorexigenic action of CRF has not been confirmed in adult bullfrogs. In this context, we examined the effect of feeding status on the expression level of the CRF transcript in the hypothalamus of the adult bullfrog. Levels of CRF mRNA in the hypothalami from bullfrogs fasted for 7 days were lower than in those from the bullfrogs that had been fed normally. Subsequently, we developed a method for measuring food intake in adult bullfrogs, and then investigated the effect of CRF on their food consumption in these animals. Intracerebroventricular (ICV) administration of CRF at 1 and 10pmol/g body weight (BW) induced a significant decrease of food intake during 60min. The CRF-induced anorexigenic action was blocked by treatment with a CRF receptor 1/CRF receptor 2 antagonist, α-helical CRF((9-41)), at 100pmol/g BW. These results provide direct evidence for the inhibitory effect of CRF on food intake, and suggest the involvement of CRF in the regulation of feeding through a CRF receptor-signaling pathway in the adult bullfrog.