RESUMEN
The pathophysiology of Parkinson's disease (PD) and L-DOPA-induced dyskinesia (LID) is associated with aberrant neuronal activity and abnormal high levels of oscillatory activity and synchronization in several basal ganglia nuclei and the cortex. Previously, we have shown that the firing activity of neurons in the substantia nigra pars reticulata (SNr) is relevant in dyskinesia and may be driven by subthalamic nucleus (STN) hyperactivity. Conversely, low frequency oscillatory activity and synchronization in these structures seem to be more important in PD because they are not influenced by prolonged L-DOPA administration. The aim of the present study was to assess (through single-unit extracellular recording techniques under urethane anaesthesia) the neuronal activity of the entopeduncular nucleus (EPN) and its relationship with LID and STN hyperactivity, together with the oscillatory activity and synchronization between these nuclei and the cerebral cortex in 6-OHDA-lesioned rats that received long term L-DOPA treatment (or not). Twenty-four hours after the last L-DOPA injection the firing activity of EPN neurons in long term L-DOPA treated 6-OHDA-lesioned rats was more irregular and bursting compared to sham rats, being those alterations partially reversed by the acute challenge of L-DOPA. No correlation between EPN neurons firing activity and abnormal involuntary movements score was found. However, there was a significant correlation between the firing activity parameters of EPN and STN neurons recorded from long term L-DOPA treated 6-OHDA-lesioned rats. Low frequency oscillatory activity and synchronization both within the EPN and with the cerebral cortex were enhanced in 6-OHDA-lesioned animals. These changes were reversed by the acute L-DOPA challenge only in long term L-DOPA treated 6-OHDA-lesioned rats. Altogether, these results obtained from long term L-DOPA treated 6-OHDA-lesioned rats suggest (1) a likely relationship between STN and EPN firing patterns and spiking phases induced by changes after prolonged L-DOPA administration and (2) that the effect of L-DOPA on the firing pattern, low frequency oscillatory activity and synchronization in the EPN may have a relevant role in LID.
Asunto(s)
Discinesia Inducida por Medicamentos/fisiopatología , Núcleo Entopeduncular/efectos de los fármacos , Núcleo Entopeduncular/fisiopatología , Levodopa/farmacología , Trastornos Parkinsonianos/fisiopatología , Adrenérgicos/toxicidad , Animales , Antiparkinsonianos/farmacología , Masculino , Neuronas/efectos de los fármacos , Oxidopamina/toxicidad , Trastornos Parkinsonianos/inducido químicamente , Ratas , Ratas Sprague-Dawley , Núcleo Subtalámico/efectos de los fármacos , Núcleo Subtalámico/fisiopatologíaRESUMEN
The authors found a terrible mistake in the manuscript. The legends from the Fig. 5 and 6 are interchanged. The Fig. 5 should be appeared with the legend from the Fig. 6 and Fig. 6 should be appeared with the legend from the Fig. 5.
RESUMEN
Rearing in enriched environment (EE) improves the recuperation in animal models of Parkinson's disease (PD). Administration of TiO2-nanowired cerebrolysin (CBL) could represent an additional strategy to protect or repair the nigrostriatal system. This study aims to explore morphofunctional and biochemical changes in a preclinical stage of PD testing the synergistic efficiency of combining both strategies, housing in EE, and nanodelivery of CBL. Sprague-Dawley male rats receiving intrastriatally 6-hydroxydopamine after a short evolution time were segregated into CBL group (rats receiving nanowired CBL), EE group (rats housed in EE), CBL + EE group (rats housed in EE and receiving nanowired CBL), and control group (rats without additional treatment). Prodromic stage and treatment effects were characterized by the presence of motor symptoms (amphetamine-induced rotational behavior test). Tyrosine hydroxylase (TH) immunohistochemistry and Western blot (p-Akt/Akt and p-ERK/ERK 1/2 as survival markers and caspase-3 as apoptotic marker) were performed in striatum and SN. A decrease in motor symptoms was shown by rats receiving CBL. EE monitoring cages revealed that rats from CBL + EE group showed more significant number of laps in the wheel than EE group. In SN, CBL + EE group also presented the highest neuronal density. Moreover, p-Akt/Akt and p-ERK/ERK 1/2 ratio was significant higher and caspase-3 expression was lower in CBL + EE group. In conclusion, the combination of CBL and EE provided evidence of neuoprotective-neurorestorative mechanisms by which this combined strategy promoted morphofunctional improvement by activation of survival pathways after dopamine depletion in a preclinical model of PD.
Asunto(s)
Aminoácidos/administración & dosificación , Modelos Animales de Enfermedad , Ambiente , Nanopartículas/administración & dosificación , Fármacos Neuroprotectores/administración & dosificación , Trastornos Parkinsonianos/tratamiento farmacológico , Animales , Sistemas de Liberación de Medicamentos/métodos , Masculino , Trastornos Parkinsonianos/metabolismo , Trastornos Parkinsonianos/patología , Ratas , Ratas Sprague-DawleyRESUMEN
Neurotrophic factors (NTFs) are a promising therapeutic option for Parkinson's disease (PD). They exert their function through tyrosine kinase receptors. Our goal was to assess the effects of administering a selective tyrosine kinase inhibitor (vandetanib) that blocks VEGFR2 and RET receptors in a preclinical model of PD. Rats underwent intrastriatal injections of 6-hydroxydopamine (6-OHDA). Two weeks later, the rats received 30 mg/kg vandetanib or saline orally. The effects were assessed using the rotational behavioral test, tyrosine hydroxylase (TH) immunohistochemistry, and western blot. In 6-OHDA-lesioned rats, motor symptoms were almost undetectable, but morphological and biochemical changes were significant. Vandetanib treatment, combined with the presence of 6-OHDA lesions, significantly increased behavioral impairment and morphological and biochemical changes. Therefore, after vandetanib treatment, the TH-immunopositive striatal volume, the percentage of TH+ neurons, and the extent of the axodendritic network in the substantia nigra decreased. Glial fibrillary acidic protein-positivity significantly decreased in the striatum and substantia nigra in the vandetanib-treated group. In addition, p-Akt and p-ERK 1/2 levels were significantly lower and caspase-3 expression significantly increased after vandetanib administration. In conclusion, we demonstrate for the first time the deleterious effect of a tyrosine kinase inhibitor on the dopaminergic system, supporting the beneficial and synergistic effect of NTFs reported in previous papers.
Asunto(s)
Trastornos Parkinsonianos/metabolismo , Piperidinas/toxicidad , Proteínas Proto-Oncogénicas c-ret/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-ret/metabolismo , Quinazolinas/toxicidad , Receptor 2 de Factores de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Receptor 2 de Factores de Crecimiento Endotelial Vascular/metabolismo , Animales , Masculino , Trastornos Parkinsonianos/inducido químicamente , Trastornos Parkinsonianos/patología , Ratas , Ratas Sprague-Dawley , Resultado del TratamientoRESUMEN
The unilateral 6-hydroxydopamine (6-OHDA) lesion of medial forebrain bundle (MFB) in rats affords us to study the advanced stages of Parkinson's disease (PD). Numerous evidences suggest synergic effects when various neurotrophic factors are administered in experimental models of PD. The aim of the present work was to assess the morphological changes along the rostro-caudal axis of caudo-putamen complex and substantia nigra (SN) in the referred model in order to test the suitability of a severe model to evaluate new neurorestorative therapies. Administration of 6-OHDA into MFB in addition to a remarkable depletion of dopamine in the nigrostriatal system induced an increase of glial fibrillary acidic protein (GFAP)-positive cells in SN and an intense immunoreactivity for OX-42, vascular endothelial growth factor (VEGF), and Lycopersycum esculentum agglutinin (LEA) in striatum and SN. Tyrosine hydroxylase (TH) immunostaining revealed a significant decrease of the TH-immunopositive striatal volume in 6-OHDA group from rostral to caudal one. The loss of TH-immunoreactive (TH-ir) neurons and axodendritic network (ADN) was higher in caudal sections. Morphological recovery after the implantation of microspheres loaded with VEGF and glial cell line-derived neurotrophic factor (GDNF) in parkinsonized rats was related to the preservation of the TH-ir cell number and ADN in the caudal region of the SN. In addition, these findings support the neurorestorative role of VEGF+GDNF in the dopaminergic system and the synergistic effect between both factors. On the other hand, a topological distribution of the dopaminergic system was noticeable in the severe model, showing a selective vulnerability to 6-OHDA and recovering after treatment.
Asunto(s)
Composición de Medicamentos , Factor Neurotrófico Derivado de la Línea Celular Glial/administración & dosificación , Trastornos Parkinsonianos/tratamiento farmacológico , Trastornos Parkinsonianos/patología , Índice de Severidad de la Enfermedad , Factor A de Crecimiento Endotelial Vascular/administración & dosificación , Animales , Composición de Medicamentos/métodos , Femenino , Fármacos Neuroprotectores/administración & dosificación , Ratas , Ratas Sprague-Dawley , Resultado del TratamientoRESUMEN
The pathophysiology of Parkinson's disease (PD) and of L-DOPA-induced dyskinesia (LID) is associated with dysfunctional neuronal activity in several nuclei of the basal ganglia. Moreover, high levels of oscillatory activity and synchronization have also been described in both intra- and inter-basal ganglia nuclei and the cerebral cortex. However, the relevance of these alterations in the motor symptomatology related to Parkinsonism and LID is not fully understood. Recently, we have shown that subthalamic neuronal activity correlates with axial abnormal movements and that a subthalamic nucleus (STN) lesion partially reduces LID severity as well as the expression of some striatal molecular modifications. The aim of the present study was to assess, through single-unit extracellular recording techniques under urethane anaesthesia, neuronal activity of the substantia nigra pars reticulata (SNr) and its relationship with LID and STN hyperactivity together with oscillatory and synchronization between these nuclei and the cerebral cortex in 6-OHDA-lesioned and dyskinetic rats. Twenty-four hours after the last injection of L-DOPA the firing rate and the inhibitory response to an acute challenge of L-DOPA of SNr neurons from dyskinetic animals were increased with respect to those found in intact and 6-OHDA-lesioned rats. Moreover, there was a significant correlation between the mean firing rate of SNr neurons and the severity of the abnormal movements (limb and orolingual subtypes). There was also a significant correlation between the firing activity of SNr and STN neurons recorded from dyskinetic rats. In addition, low frequency band oscillatory activity and synchronization both within the SNr or STN and with the cerebral cortex were enhanced in 6-OHDA-lesioned animals and not or slightly affected by chronic treatment with L-DOPA. Altogether, these results indicate that neuronal SNr firing activity is relevant in dyskinesia and may be driven by STN hyperactivity. Conversely, low frequency oscillatory activity and synchronization seem to be more important in PD because they are not influenced by prolonged L-DOPA administration.
Asunto(s)
Potenciales de Acción/efectos de los fármacos , Ondas Encefálicas/efectos de los fármacos , Encéfalo/fisiopatología , Discinesia Inducida por Medicamentos/fisiopatología , Levodopa/administración & dosificación , Neuronas/fisiología , Trastornos Parkinsonianos/fisiopatología , Animales , Encéfalo/efectos de los fármacos , Corteza Cerebral/efectos de los fármacos , Corteza Cerebral/fisiopatología , Modelos Animales de Enfermedad , Neuronas GABAérgicas/efectos de los fármacos , Neuronas GABAérgicas/fisiología , Neuronas/efectos de los fármacos , Oxidopamina/administración & dosificación , Trastornos Parkinsonianos/inducido químicamente , Porción Reticular de la Sustancia Negra/efectos de los fármacos , Porción Reticular de la Sustancia Negra/fisiopatología , Ratas , Ratas Sprague-Dawley , Núcleo Subtalámico/efectos de los fármacos , Núcleo Subtalámico/fisiopatologíaRESUMEN
Parkinson's disease (PD) is the second most frequent neurodegenerative disorder, but current therapies are only symptomatic. A promising alternative to address the neurodegenerative process is the use of neurotrophic factors, such as the glial cell-derived neurotrophic factor (GDNF). However, its clinical use has been limited due to its short half-life and rapid degradation after in vivo administration, in addition to difficulties in crossing the blood-brain barrier (BBB). This barrier is a limiting factor in brain drug development, making the future progression of neurotherapeutics difficult. In the past few years, intranasal drug delivery has appeared as an alternative non-invasive administration route to bypass the BBB and target drugs directly to the CNS. Thus, the aim of this work was to study the in vivo neuroprotective effect of intranasally administered GDNF, encapsulated in chitosan-coated nanostructured lipid carrier (CS-NLC-GDNF), in a 6-OHDA partially lesioned rat model. The developed CS-NLC-GDNF showed a particle size of approximately 130 nm and high encapsulation efficiency. The in vitro study in PC-12 cells demonstrated the ability of the encapsulated GDNF to protect these cells against 6-OHDA toxin. After two weeks of daily intranasal administration of treatments, the administration of CS-NLC-GDNF achieved a behavioral improvement in rats, as well as a significant improvement in both the density of TH+ fibres in the striatum and the TH+ neuronal density in the SN. Thus, it can be concluded that the nose-to-brain delivery of CS-NLC-GDNF could be a promising therapy for the treatment of PD.
Asunto(s)
Portadores de Fármacos/química , Factor Neurotrófico Derivado de la Línea Celular Glial/farmacología , Lípidos/química , Nanoestructuras/química , Enfermedad de Parkinson/metabolismo , Administración Intranasal , Animales , Conducta Animal/efectos de los fármacos , Cuerpo Estriado/química , Cuerpo Estriado/efectos de los fármacos , Modelos Animales de Enfermedad , Portadores de Fármacos/administración & dosificación , Factor Neurotrófico Derivado de la Línea Celular Glial/administración & dosificación , Factor Neurotrófico Derivado de la Línea Celular Glial/química , Lípidos/administración & dosificación , Masculino , Nanoestructuras/administración & dosificación , Células PC12 , Tamaño de la Partícula , Ratas , Ratas Sprague-DawleyRESUMEN
The most effective treatment for Parkinson's disease (PD), l-DOPA, induces dyskinesia after prolonged use. We have previously shown that in 6-hydroxydopamine (6-OHDA) lesioned rats rendered dyskinetic by prolonged l-DOPA administration, lesion of the subthalamic nucleus (STN) reduces not only dyskinesias but also buspirone antidyskinetic effect. This study examined the effect of buspirone on STN neuron activity. Cell-attached recordings in parasagittal slices from naïve rats showed that whilst serotonin excited the majority of STN neurons, buspirone showed an inhibitory main effect but only in 27% of the studied cells which was prevented by the 5-HT1A receptor selective antagonist WAY-100635. Conversely, single-unit extracellular recordings were performed in vivo on STN neurons from four different groups, i.e., control, chronically treated with l-DOPA, 6-OHDA lesioned and lesioned treated with l-DOPA (dyskinetic) rats. In control animals, systemic-buspirone administration decreased the firing rate in a dose-dependent manner in every cell studied. This effect, prevented by WAY-100635, was absent in 6-OHDA lesioned rats and was not modified by prolonged l-DOPA administration. Altogether, buspirone in vivo reduces consistently the firing rate of the STN neurons through 5-HT1A receptors whereas ex vivo buspirone seems to affect only a small population of STN neurons. Furthermore, the lack of effect of buspirone in 6-OHDA lesioned rats, suggests the requirement of not only the activation of 5-HT1A receptors but also an intact nigrostriatal pathway for buspirone to inhibit the STN activity.
Asunto(s)
Buspirona/farmacología , Neuronas/efectos de los fármacos , Receptor de Serotonina 5-HT1A/metabolismo , Agonistas de Receptores de Serotonina/farmacología , Núcleo Subtalámico/citología , Potenciales de Acción/efectos de los fármacos , Adrenérgicos/toxicidad , Inhibidores de Captación Adrenérgica/farmacología , Animales , Desipramina/farmacología , Modelos Animales de Enfermedad , Dopaminérgicos/efectos adversos , Discinesia Inducida por Medicamentos/etiología , Discinesia Inducida por Medicamentos/prevención & control , Femenino , Levodopa/efectos adversos , Masculino , Síndromes de Neurotoxicidad/tratamiento farmacológico , Síndromes de Neurotoxicidad/etiología , Oxidopamina/toxicidad , Ratas , Ratas Sprague-DawleyRESUMEN
Administration of various neurotrophic factors is a promising strategy against Parkinson's disease (PD). An intrastriatal infusion of 6-hydroxidopamine (6-OHDA) in rats is a suitable model to study PD. This work aims to describe stereological parameters regarding rostro-caudal gradient, in order to characterize the model and verify its suitability for elucidating the benefits of therapeutic strategies. Administration of 6-OHDA induced a reduction in tyrosine hidroxylase (TH) reactivity in the dorsolateral part of the striatum, being higher in the caudal section than in the rostral one. Loss of TH-positive neurons and axodendritic network was highly significant in the external third of substantia nigra (e-SN) in the 6-OHDA group versus the saline one. After the administration of nanospheres loaded with neurotrophic factors (NTF: vascular endothelial growth factor (VEGF) + glial cell line-derived neurotrophic factor (GDNF)), parkinsonized rats showed more TH-positive fibers than those of control groups; this recovery taking place chiefly in the rostral sections. Neuronal density and axodendritic network in e-SN was more significant than in the entire SN; the topographical analysis showed that the highest difference between NTF versus control group was attained in the middle section. A high number of bromodeoxyuridine (BrdU)-positive cells were found in sub- and periventricular areas in the group receiving NTF, where most of them co-expressed doublecortin. Measurements on the e-SN achieved more specific and significant results than in the entire SN. This difference in rostro-caudal gradients underpins the usefulness of a topological approach to the assessment of the lesion and therapeutic strategies. Findings confirmed the neurorestorative, neurogenic, and synergistic effects of VEGF+GDNF administration.
Asunto(s)
Factor Neurotrófico Derivado de la Línea Celular Glial/uso terapéutico , Trastornos Parkinsonianos/tratamiento farmacológico , Factor A de Crecimiento Endotelial Vascular/uso terapéutico , Animales , Cuerpo Estriado/efectos de los fármacos , Cuerpo Estriado/enzimología , Replicación del ADN , Proteínas de Dominio Doblecortina , Proteína Doblecortina , Sinergismo Farmacológico , Quimioterapia Combinada , Factor Neurotrófico Derivado de la Línea Celular Glial/administración & dosificación , Factor Neurotrófico Derivado de la Línea Celular Glial/farmacocinética , Proteína Ácida Fibrilar de la Glía/análisis , Inyecciones Intralesiones , Masculino , Proteínas Asociadas a Microtúbulos/análisis , Nanocápsulas , Proteínas del Tejido Nervioso/análisis , Neuroglía/química , Neuroglía/efectos de los fármacos , Neuroglía/ultraestructura , Neuronas/química , Neuronas/efectos de los fármacos , Neuronas/ultraestructura , Neuropéptidos/análisis , Oxidopamina/toxicidad , Trastornos Parkinsonianos/patología , Ratas , Ratas Sprague-Dawley , Distribución Tisular , Tirosina 3-Monooxigenasa/análisis , Factor A de Crecimiento Endotelial Vascular/administración & dosificación , Factor A de Crecimiento Endotelial Vascular/farmacocinéticaRESUMEN
RATIONALE: Within the basal ganglia, the subthalamic nucleus (STN) is the only glutamatergic structure and occupies a central position in the indirect pathway. In rat, the STN receives serotonergic input from the dorsal raphe nucleus and expresses serotonergic receptors. OBJECTIVE: This study examined the consequences of serotonergic neurotransmission modulation on STN neuron activity. METHODS: In vivo single-unit extracellular recordings, HPLC determination, and rotarod and bar test were performed in control, 4-chloro-DL-phenylalanine methyl ester hydrochloride- (pCPA, a serotonin synthesis inhibitor) and chronically fluoxetine-treated rats. RESULTS: The pCPA treatment and the administration of serotonin (5-HT) receptor antagonists increased number of bursting neurons in the STN. The systemic administration of the 5-HT(1A) agonist, 8-OH-DPAT, decreased the firing rate and increased the coefficient of variation of STN neurons in pCPA-treated rats but not in control animals. Additionally, microinjection of 8-OH-DPAT into the STN reduced the firing rate of STN neurons, while microinjection of the 5-HT(2C) agonist, Ro 60-0175, increased the firing rate in both control and fluoxetine-treated animals. Finally, the fluoxetine challenge increased the firing rate of STN neurons in fluoxetine-treated rats and induced catalepsy. CONCLUSIONS: Our results indicate that the depletion and the blockage of 5-HT modify STN neuron firing pattern. STN neuron activity is under the control of 5-HT(1A) and 5-HT(2C) receptors located both inside and outside the STN. Finally, fluoxetine increases STN neuron activity in chronically fluoxetine-treated rats, which may explain the role of this nucleus in fluoxetine-induced extrapyramidal side effects.
Asunto(s)
Fenclonina/análogos & derivados , Fluoxetina/farmacología , Neuronas/efectos de los fármacos , Inhibidores Selectivos de la Recaptación de Serotonina/farmacología , Serotoninérgicos/farmacología , Núcleo Subtalámico/efectos de los fármacos , 8-Hidroxi-2-(di-n-propilamino)tetralin/farmacología , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/fisiología , Animales , Etilaminas/farmacología , Fenclonina/farmacología , Indoles/farmacología , Masculino , Actividad Motora/efectos de los fármacos , Actividad Motora/fisiología , Neuronas/fisiología , Ratas Sprague-Dawley , Receptor de Serotonina 5-HT1A/metabolismo , Receptor de Serotonina 5-HT2C/metabolismo , Agonistas del Receptor de Serotonina 5-HT1/farmacología , Agonistas del Receptor de Serotonina 5-HT2/farmacología , Antagonistas de la Serotonina/farmacología , Núcleo Subtalámico/fisiologíaRESUMEN
Parkinson's disease (PD) is a multisystemic disorder in which several neurotransmitters other than dopamine are affected. Drugs acting on non-dopaminergic systems are envisaged as promising agents to treat PD and levodopa-induced dyskinesias (LID). However, compounds targeting glutamate, adenosine, noradrenaline, 5-hydroxytryptamine, cannabinoid, and opioid transmitter systems have been assessed in human studies showing negative, inconsistent or unsatisfactory results. Most of these drugs had been tested previously in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-lesioned monkeys, as well as in the classic 6-hydroxydopamine-lesioned rat model. These failures raise several questions and concerns about the true reliability of animal studies, the adequacy of the working hypotheses and design of clinical trials, the validity of tools in current use to evaluate a particular effect, and the selectivity of the drugs used. More importantly, observed discrepancies between the results in models and patients, could challenge the validity of current ideas about the pathophysiology of parkinsonism and LID.
Asunto(s)
Antiparkinsonianos/farmacología , Neurotransmisores , Enfermedad de Parkinson/tratamiento farmacológico , Receptores de Dopamina D2/efectos de los fármacos , Receptores de Dopamina D3/efectos de los fármacos , Animales , Ensayos Clínicos como Asunto , Dopaminérgicos/farmacología , HumanosRESUMEN
The cannabinoid CB1 receptor which is densely located in the basal ganglia is known to participate in the regulation of movement. The present study sought to determine the mechanisms underlying the effect of Delta(9)-tetrahydrocannabinol (Delta(9)-THC) on neurons in the substantia nigra pars compacta (SNpc) using single-unit extracellular recordings in anesthetized rats. Administration of Delta(9)-THC (0.25-2 mg/kg, i.v.) increased the firing rate of SNpc neurons (maximal effect: 33.54+/-6.90%, n=8) without modifying other firing parameters (coefficient of variation and burst firing). This effect was completely blocked by the cannabinoid receptor antagonist rimonabant (0.5 mg/kg, i.v.). In addition, the blockade of excitatory amino acids receptors by kynurenic acid (0.5 microM, i.c.v.) or a chemical lesion of the subthalamic nucleus (STN) with ibotenic acid abolished Delta(9)-THC effect. These results indicate that CB1 receptor activation modulates SNpc neuronal activity by an indirect mechanism involving excitatory amino acids, probably released from STN axon terminals in the SNpc.
Asunto(s)
Dopamina/metabolismo , Dronabinol/farmacología , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Psicotrópicos/farmacología , Núcleo Subtalámico/fisiología , Potenciales de Acción/efectos de los fármacos , Análisis de Varianza , Animales , Apomorfina/farmacología , Agonistas de Dopamina/farmacología , Relación Dosis-Respuesta a Droga , Agonistas de Aminoácidos Excitadores/efectos adversos , Ácido Iboténico/efectos adversos , Ácido Quinurénico/efectos adversos , Masculino , Piperidinas/farmacología , Pirazoles/farmacología , Ratas , Ratas Sprague-Dawley , Rimonabant , Sustancia Negra/citología , Núcleo Subtalámico/lesionesRESUMEN
The in vivo effect of the serotonin (5-HT) reuptake inhibitor antidepressant citalopram, administered in the locus coeruleus (LC), on noradrenergic transmission was evaluated in the rat brain. In dual-probe microdialysis assays, citalopram (0.1-100 microM), in a concentration-dependent manner, increased extracellular noradrenaline (NA) in the LC and simultaneously decreased extracellular NA in the cingulate cortex (Cg). These effects of citalopram were abolished by pretreatment with the 5-HT synthesis inhibitor p-chlorophenylalanine (400 mg/kg, i.p.). When the alpha(2)-adrenoceptor antagonist RS79948 (1 microM) was perfused in the LC, local citalopram increased NA dialysate in the LC but no longer modified NA dialysate in the Cg. In electrophysiological experiments, the administration of citalopram (100 microM) in the LC by reversal dialysis, decreased the firing rate of LC neurones. The results demonstrate in vivo that local administration of citalopram in the LC leads to a decreased release of NA in the Cg. This modulation seems to be the result of an increase in NA concentration in the LC and the subsequent inhibition of LC neurones via alpha(2)-adrenoceptors. The effects of citalopram are dependent on the presence of endogenous 5-HT in the LC.
Asunto(s)
Citalopram/farmacología , Locus Coeruleus/efectos de los fármacos , Norepinefrina/metabolismo , Inhibidores Selectivos de la Recaptación de Serotonina/farmacología , Serotonina/metabolismo , Animales , Locus Coeruleus/metabolismo , Microdiálisis , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Ratas , Ratas Sprague-DawleyAsunto(s)
Benzofuranos/farmacología , Idazoxan/farmacología , Imidazoles/farmacología , Neuronas/fisiología , Núcleos del Rafe/fisiología , Serotonina/farmacología , Serotonina/fisiología , Animales , Clorgilina/farmacología , Receptores de Imidazolina , Técnicas In Vitro , Neuronas/efectos de los fármacos , Fenilefrina/farmacología , Ratas , Ratas Sprague-Dawley , Receptores Adrenérgicos alfa 2/fisiología , Receptores de Droga/fisiologíaRESUMEN
1. Imidazoline binding sites have been reported to be present in the locus coeruleus (LC). To investigate the role of these sites in the control of LC neuron activity, we studied the effect of imidazolines using in vivo and in vitro single-unit extracellular recording techniques. 2. In anaesthetized rats, local (27 pmoles) and systemic (1 mg kg(-1), i.v.) administrations of 2-(2-benzofuranyl)-2-imidazoline (2-BFI), a selective I-imidazoline receptor ligand, increased the firing rate of LC cells (maximal increase: 22+/-5%, P<0.001 and 16+/-7%, P<0.001 respectively). Chronic pretreatment with the irreversible monoamine oxidase inhibitor clorgyline (3 mg kg(-1), i.p., every 12 h for 14 days) abolished this effect. 3. In rat midpontine brain slices containing the LC, bath application (1 mM) of the imidazolines 2-BFI, 2-(4,5-dihydroimidaz-2-yl)-quinoline (BU224), idazoxan, efaroxan, phentolamine and (2-2-methoxy-1,4-benzodioxan-2-yl)-2-imidazoline (RX821002) reversibly stimulated LC cells. The maximal effect was approximately 90% except for RX821002 and efaroxan which induced smaller maximal effects (approximately 58% and approximately 35% respectively). Simultaneous application of idazoxan and 2BFI did not lead to additive effects. 4. Bath application of the alpha2-adrenoceptor antagonists, yohimbine (1 - 10 microM) and N-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ) (10 microM), failed to modify LC activity. The irreversible blockade of alpha2-adrenoceptors with EEDQ (10 microM) did not alter the effect of idazoxan or that of efaroxan. Previous application of clorgyline (10 microM) did not modify the excitatory effect of 2-BFI or efaroxan. 5. Changes in the pH of the bathing solution (6.84-7.84) did not influence the effect caused by idazoxan. Bath application of 2-BFI (1 mM) reversed the inhibition induced by diazoxide (300 microM), an ATP-sensitive K+ channel opener, whereas application of glibenclamide (3 microM), an ATP-sensitive K+ channel blocker, partially blocked the effect of 2-BFI. 6. This study shows that imidazoline compounds stimulate the firing rate of LC neurons. This effect is not mediated by alpha2-adrenoceptors nor by I1 or I2-imidazoline receptors but involves a different subtype of imidazoline receptor. Our results indicate that this receptor is located extracellularly and modulates ATP-sensitive K+ channels.
Asunto(s)
Locus Coeruleus/metabolismo , Neuronas/metabolismo , Receptores de Droga/metabolismo , Adenosina Trifosfato/fisiología , Antagonistas de Receptores Adrenérgicos alfa 2 , Antagonistas Adrenérgicos alfa/farmacología , Animales , Benzofuranos/farmacología , Diazóxido/farmacología , Electrofisiología , Idazoxan/farmacología , Imidazoles/farmacología , Receptores de Imidazolina , Técnicas In Vitro , Ligandos , Locus Coeruleus/efectos de los fármacos , Locus Coeruleus/fisiología , Masculino , Neuronas/efectos de los fármacos , Neuronas/fisiología , Canales de Potasio/efectos de los fármacos , Canales de Potasio/fisiología , Ratas , Receptores Adrenérgicos alfa 2/metabolismo , Receptores de Droga/efectos de los fármacos , Vasodilatadores/farmacologíaRESUMEN
Using in vivo extracellular recordings, we have examined the effect of the application of the prototypical 5-HT1A receptor agonist 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT), on the firing rate of locus coeruleus neurons. 8-OH-DPAT (1 microgram/kg, i.v.) did not modify the basal activity of the locus coeruleus but shifted to the left the dose-response curve for the clonidine induced inhibition of firing rate and reduced the corresponding ED50 by 77%. 2-[2-[4-(o-methoxyphenyl)piperazin-1-yl]ethyl]-4,4-dimethyl-1,3(2H ,4H)-isoquinolinedione (ARC 239; 75 micrograms/kg, i.v.), and chlorpromazine (75 micrograms/kg, i.v.) also shifted to the left the dose-response curve for clonidine and reduced by 38 and 46%, respectively, the ED50, while slightly increasing the basal firing rate. The results indicated that 5-HT1A receptors may modulate the responses mediated by alpha 2A-adrenoceptors in the locus coeruleus.
Asunto(s)
Agonistas alfa-Adrenérgicos/farmacología , Clonidina/farmacología , Locus Coeruleus/efectos de los fármacos , Receptores de Serotonina/efectos de los fármacos , Agonistas de Receptores de Serotonina/farmacología , 8-Hidroxi-2-(di-n-propilamino)tetralin/farmacología , Antagonistas de Receptores Adrenérgicos alfa 2 , Antagonistas Adrenérgicos alfa/farmacología , Animales , Clorpromazina/farmacología , Antagonistas de Dopamina/farmacología , Relación Dosis-Respuesta a Droga , Inyecciones Intravenosas , Isoquinolinas/farmacología , Locus Coeruleus/metabolismo , Masculino , Piperazinas/farmacología , Ratas , Ratas Sprague-Dawley , Receptores de Serotonina/metabolismo , Receptores de Serotonina 5-HT1RESUMEN
The population of reserve alpha-2 adrenoceptors that mediate the inhibitory effect of clonidine on the activity of locus coeruleus neurons has been studied using extracellular recordings in anesthetized rats. Animals were pretreated with the irreversible receptor antagonist N-ethoxycarbonyl-2-ethoxy-1-2-dihydroquinoline (EEDQ). In rats pretreated with EEDQ (1, 2 and 6 mg/kg, i.p., 6 hr before experiment), there was an increase in firing rate, a reduction in firing regularity (i.e., increased variation coefficient) and an increase in burst firing of locus coeruleus neurons. Partial receptor inactivation with EEDQ (1 and 2 mg/kg, i.p.) caused a dose-dependent shift to the right of dose-effect curves for i.v. administered clonidine together with a reduction in its maximal effect. Higher doses of EEDQ (6 mg/kg, i.p.) completely abolished the effect induced by clonidine. This blockade was associated with a progressive decrease in the number of remaining receptors (noninactivated receptors). The pseudo-constant of dissociation for the drug-receptor complex was calculated to be approximately 70 micrograms/kg. The receptor occupancy-effect relationship was hyperbolic giving a value of only approximately 4% occupancy at 50% maximal effect. Estimates of noninactivated receptors and percentage of receptor occupancy at 50% of maximal effect were comparable when locally administered clonidine was used. After complete receptor inactivation with EEDQ (6 mg/kg), dose-effect curves for clonidine recovered gradually. The inhibitory effect of clonidine returned faster (half-life = 14 hr) than the receptor pool (half-life = 37 hr). These results indicate that locus coeruleus neurons have a large reserve of alpha 2 adrenoceptors that in addition, are rapidly turned over.
Asunto(s)
Agonistas alfa-Adrenérgicos/farmacología , Clonidina/farmacología , Locus Coeruleus/efectos de los fármacos , Neuronas/efectos de los fármacos , Receptores Adrenérgicos alfa 2/metabolismo , Antagonistas de Receptores Adrenérgicos alfa 2 , Antagonistas Adrenérgicos alfa/farmacología , Animales , Locus Coeruleus/citología , Locus Coeruleus/metabolismo , Masculino , Neuronas/metabolismo , Quinolinas/farmacología , Ratas , Ratas Sprague-Dawley , Receptores Adrenérgicos alfa 2/fisiologíaRESUMEN
Clonidine stimulates locus coeruleus neurons by an alpha 2-adrenoceptor-independent mechanism which probably involves imidazoline receptors. To study this effect, single-unit extracellular recordings in the locus coeruleus were performed in anaesthetised rats after complete, irreversible inactivation of alpha 2-adrenoceptors by the alkylating agent N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) (6 mg/kg i.p.; 6 h before experiments). After this pretreatment, clonidine applied into the locus coeruleus failed to produce any change in the cell firing rate. However, clonidine applied intravenously (320-2560 micrograms/kg), or locally (0.5-2.0 microliters of 0.02 M) into the nucleus paragigantocellularis, a major locus coeruleus afferent, stimulated locus coeruleus neurons (increasing the firing rate by approximately 90%). Electrical lesions of the nucleus paragigantocellularis greatly attenuated the clonidine induced stimulation of locus coeruleus neurons ipsilateral to the lesion when applied intravenously. Blood pressure which was recorded simultaneously with cell recording, remained unaffected after clonidine administration in EEDQ pretreated rats. These results indicate that the clonidine-induced stimulation of locus coeruleus neurons is an indirect effect mediated by imidazoline receptors located on paragigantocellularis neurons projecting to the locus coeruleus.
Asunto(s)
Antagonistas de Receptores Adrenérgicos alfa 2 , Agonistas alfa-Adrenérgicos/farmacología , Clonidina/farmacología , Locus Coeruleus/efectos de los fármacos , Neuronas/efectos de los fármacos , Receptores de Droga/metabolismo , Antagonistas Adrenérgicos alfa/farmacología , Animales , Presión Sanguínea/efectos de los fármacos , Clonidina/administración & dosificación , Electrofisiología , Receptores de Imidazolina , Locus Coeruleus/fisiología , Locus Coeruleus/ultraestructura , Masculino , Bulbo Raquídeo/metabolismo , Microinyecciones , Neuronas/fisiología , Quinolinas/farmacología , Ratas , Ratas Sprague-DawleyRESUMEN
Agmatine (decarboxylated arginine) has been proposed as an endogenous ligand for non-adrenoceptor, imidazoline binding sites, but also binds to alpha 2-adrenoceptors. The interaction of agmatine with alpha 2-adrenoceptors was evaluated by studying the effect of agmatine on the firing rate of locus coeruleus (LC) neurones using extracellular recordings in anesthetized rats and rat brain slices. In vivo, local application of agmatine into the LC caused a slight and short-lasting increase in cell firing rate (P < 0.005). In vitro, agmatine failed to change the firing rate of LC neurones nor did it antagonize the inhibitory effect of noradrenaline on these cells. Since alpha 2-adrenoceptors are known to inhibit the firing of LC cells, we conclude that agmatine does not have agonist or antagonist properties at alpha 2-adrenoceptors of these neurones.
Asunto(s)
Agmatina/metabolismo , Locus Coeruleus/fisiología , Neuronas/fisiología , Receptores Adrenérgicos alfa/fisiología , Agmatina/farmacología , Animales , Electrofisiología , Técnicas In Vitro , Locus Coeruleus/citología , Locus Coeruleus/efectos de los fármacos , Masculino , Neuronas/efectos de los fármacos , Norepinefrina/farmacología , Ratas , Ratas Sprague-Dawley , Receptores Adrenérgicos alfa/efectos de los fármacosRESUMEN
Clonidine and other imidazoline/oxazoline drugs, such as cirazoline and rilmenidine, have been shown to stimulate the activity of noradrenergic neurones in the locus coeruleus (NA-LC) by an alpha 2-adrenoceptor-independent mechanism through the activation of I-imidazoline receptors. The endogenous modulation of the stimulatory effect of clonidine on NA-LC neurones was further investigated after inactivation of alpha 2-adrenoceptors with N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ). In EEDQ-pretreated rats (6 mg/kg, i.p., 6 h), clonidine caused a rapid and dose-dependent (320-5120 micrograms/kg, i.v.) increase in the firing rate of NA-LC neurones (ED50 = 809 micrograms/kg, Emax = 90%). The stimulatory effect of clonidine on NA-LC neurones was completely blocked by pretreatment of rats with the excitatory amino acid receptor antagonist kynurenic acid (1-3 mumol in 10-30 microliters, i.c.v., 2-5 min before clonidine). In contrast, the stimulatory effect of clonidine on NA-LC neurones was potentiated by pretreatment with reserpine (5 mg/kg, s.c., 18 h) (Emax increased by 63%). Pretreatment with alpha-methyl-p-tyrosine (250 mg/kg, i.p., 24 h) did not alter the stimulatory effect of clonidine, but pretreatment with p-chloro-phenylalanine (400 mg/kg, i.p., 24 h) markedly enhanced the stimulatory effect of clonidine on NA-LC neurones (Emax increased by 139%). The present results indicate that the imidazoline receptor-mediated stimulatory effect of clonidine on NA-LC neurones is an indirect effect dependent on an excitatory amino acid pathway and modulated by an inhibitory serotonin mechanism.