RESUMEN
An empirical model for the evolution of ß-phase (Mg2Al3) along grain boundaries in aluminium alloy AA5083 (Al-Mg-Mn) during isothermal exposures is proposed herein. Developing a quantitative understanding of grain boundary precipitation is important to interpreting intergranular corrosion and stress corrosion cracking in this alloy system. To date, complete ab initio models for grain boundary precipitation based upon fundamental principles of thermodynamics and kinetics are not available, despite the critical role that such precipitates play in dictating intergranular corrosion phenomena. Empirical models can therefore serve an important role in advancing the understanding of grain boundary precipitation kinetics, which is an approach applicable beyond the present context. High resolution scanning electron microscopy was to quantify the size and distribution of ß-phase precipitates on Ga-embrittled intergranular fracture surfaces of AA5083. The results are compared with the degree of sensitisation (DoS) as judged by nitric acid mass loss testing (ASTM-G67-04), and discussed with models for sensitisation in 5xxx series Al-alloys. The work herein allows sensitisation to be quantified from an unambiguous microstructural perspective.
RESUMEN
Clinically, amantadine and memantine are drugs whose therapeutic utility is linked to their ability to block N-methyl-D-aspartate receptors (NMDARs) in a voltage-dependent manner. Nevertheless many studies that have characterized the pharmacological actions of amantadine and memantine have done so in the absence of physiological levels of Mg(2+) ions. This study quantifies the extent to which Mg(2+) alters the potency of the block produced by both amantadine and memantine at human recombinant GluN1/GluN2A NMDARs. Human recombinant GluN1/GluN2A NMDARs were expressed in Xenopus laevis oocytes and two-electrode voltage-clamp recordings were made at -80, -60 and -40 mV to quantify amantadine and memantine block in the absence and presence of Mg(2+). Amantadine and memantine blocked human GluN1/GluN2A NMDARs in a voltage-dependent manner with IC(50) values (at -80 mV) of 49 ± 6 µM (n = 7) and 1.0 ± 0.3 µM (n = 7), respectively. In the presence of Mg(2+) (1mM) the equivalent IC(50) values were 165 ± 10 µM (n=6) and 6.6 ± 0.3 µM (n = 5). Similarly in the presence of amantadine or memantine the potency of Mg(2+) in blocking GluN1/GluN2A NMDARs was reduced. The decrease in the potencies of both amantadine and memantine in the presence of physiological concentrations of Mg(2+) indicates that other targets (e.g. α7-nicotinic acetylcholine receptors and 5-HT(3) receptors) in addition to NMDARs may well be sites of the therapeutic action of these channel blockers.
Asunto(s)
Amantadina/farmacología , Magnesio/farmacología , Memantina/farmacología , Receptores de N-Metil-D-Aspartato/antagonistas & inhibidores , Animales , Relación Dosis-Respuesta a Droga , Interacciones Farmacológicas , Femenino , Humanos , Receptores de N-Metil-D-Aspartato/fisiología , Proteínas Recombinantes/antagonistas & inhibidores , Xenopus laevisRESUMEN
One of the major neuropathological hallmarks in Alzheimer's disease (AD) is the loss of cholinergic neurones of the nucleus basalis of Meynert (NbM). This consistent finding gave rise to the 'cholinergic' hypothesis of AD and lead to the subsequent development of acetylcholinesterase (AChE) inhibitors; the first class of drug to be approved for the treatment of AD. However, several studies have questioned the efficacy of using AChE inhibitors in AD. In this study we have investigated the ability of two AChE inhibitors, donepezil (Aricept) and physostigmine, to induce gamma oscillatory activity in rat hippocampal slices; network activity believed to play a role in higher cognitive function. We report here that donepezil is capable of inducing gamma oscillations in region CA3 of rat hippocampal slices, which may contribute to its procognitive action. However, donepezil-induced gamma oscillations are weak in comparison to physostigmine. We also explore the activity of novel agents with known procognitive activity, and show that one such agent, the M(1) muscarinic acetylcholine receptor agonist, 77-LH-28-1, can significantly enhance donepezil-induced gamma oscillations. These data support the notion that it should be possible to find a more efficacious AChE inhibitor or an adjunctive approach, to provide a better therapeutic intervention in AD.
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Ondas Encefálicas/efectos de los fármacos , Hipocampo/efectos de los fármacos , Indanos/farmacología , Neuronas/efectos de los fármacos , Nootrópicos/farmacología , Piperidinas/farmacología , Animales , Ondas Encefálicas/fisiología , Inhibidores de la Colinesterasa/farmacología , Donepezilo , Electrofisiología , Hipocampo/fisiología , Neuronas/fisiología , Quinolonas/farmacología , RatasRESUMEN
BACKGROUND AND PURPOSE: M1 muscarinic ACh receptors (mAChRs) represent an attractive drug target for the treatment of cognitive deficits associated with diseases such as Alzheimer's disease and schizophrenia. However, the discovery of subtype-selective mAChR agonists has been hampered by the high degree of conservation of the orthosteric ACh-binding site among mAChR subtypes. The advent of functional screening assays has enabled the identification of agonists such as AC-42 (4-n-butyl-1-[4-(2-methylphenyl)-4-oxo-1-butyl]-piperidine), which bind to an allosteric site and selectively activate the M(1) mAChR subtype. However, studies with this compound have been limited to recombinantly expressed mAChRs. EXPERIMENTAL APPROACH: In this study, we have compared the pharmacological profile of AC-42 and a close structural analogue, 77-LH-28-1 (1-[3-(4-butyl-1-piperidinyl)propyl]-3,4-dihydro-2(1H)-quinolinone) at human recombinant, and rat native, mAChRs by calcium mobilization, inositol phosphate accumulation and both in vitro and in vivo electrophysiology. KEY RESULTS: Calcium mobilization and inositol phosphate accumulation assays revealed that both AC-42 and 77-LH-28-1 display high selectivity to activate the M1 mAChR over other mAChR subtypes. Furthermore, 77-LH-28-1, but not AC-42, acted as an agonist at rat hippocampal M1 receptors, as demonstrated by its ability to increase cell firing and initiate gamma frequency network oscillations. Finally, 77-LH-28-1 stimulated cell firing in the rat hippocampus in vivo following subcutaneous administration. CONCLUSIONS AND IMPLICATIONS: These data suggest that 77-LH-28-1 is a potent, selective, bioavailable and brain-penetrant agonist at the M1 mAChR and therefore that it represents a better tool than AC-42, with which to study the pharmacology of the M1 mAChR.
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Hipocampo/efectos de los fármacos , Agonistas Muscarínicos/farmacología , Piperidinas/farmacología , Quinolonas/farmacología , Receptores Muscarínicos/efectos de los fármacos , Potenciales de Acción , Animales , Células CHO , Señalización del Calcio/efectos de los fármacos , Cricetinae , Cricetulus , Relación Dosis-Respuesta a Droga , Hipocampo/metabolismo , Humanos , Inyecciones Intraperitoneales , Inyecciones Subcutáneas , Fosfatos de Inositol/metabolismo , Agonistas Muscarínicos/administración & dosificación , Agonistas Muscarínicos/farmacocinética , Técnicas de Placa-Clamp , Permeabilidad , Piperidinas/administración & dosificación , Piperidinas/farmacocinética , Quinolonas/administración & dosificación , Quinolonas/farmacocinética , Ratas , Ratas Sprague-Dawley , Receptor Muscarínico M1 , Receptores Muscarínicos/genética , Receptores Muscarínicos/metabolismo , Proteínas Recombinantes/agonistas , Factores de Tiempo , TransfecciónRESUMEN
Exposure to an enriched environment promotes neurochemical, structural and neurophysiological changes in the brain and is associated with enhanced synaptic plasticity and improved hippocampal-dependent learning. Using a global proteomics-based approach we have now been able to reveal the altered expression of a diverse range of hippocampal proteins following exposure to an enriched environment. Male Hooded Lister rats (8 weeks) were subjected to a 6-week regimen in which they were housed in either non-enriched (open field) or enriched conditions (toys, wheels etc.). Whole protein extracts from stratum pyramidale and stratum radiatum of area CA1 were then isolated and subjected to differential gel electrophoresis [McNair K, Davies CH, Cobb SR (2006) Plasticity-related regulation of the hippocampal proteome. Eur J Neurosci 23(2):575-580]. Of the 2469 resolvable protein spots detected in this study, 42 spots (1.7% of the detectable proteome) derived from predominantly somatic fractions and 32 proteins spots from dendritic fractions (1.3% of detectable proteome) were significantly altered in abundance following exposure to an enriched environment (somatic: 14 increased/28 decreased abundance, range -1.5 to +1.4-fold change; dendritic: 16 increased, 16 decreased abundance, range -1.6 to +3.0-fold change). Following in-gel tryptic digestion and Maldi-Tof/Q-star mass spectrometry, database searching revealed the identity of 50 protein spots displaying environmental enrichment-related modulation of expression. Identified proteins belonged to a variety of functional classes with gene ontology analysis revealing the majority (>70%) of regulated proteins to be part of the energy metabolism, cytoplasmic organization/biogenesis and signal transduction processes.
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Planificación Ambiental , Expresión Génica/fisiología , Hipocampo/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Plasticidad Neuronal/fisiología , Proteoma/metabolismo , Animales , Dendritas/metabolismo , Dendritas/ultraestructura , Metabolismo Energético/fisiología , Conducta Exploratoria/fisiología , Aprendizaje/fisiología , Masculino , Memoria/fisiología , Proteínas del Tejido Nervioso/análisis , Proteómica , Células Piramidales/metabolismo , Células Piramidales/ultraestructura , Ratas , Transducción de Señal/fisiología , Regulación hacia Arriba/fisiologíaRESUMEN
The hippocampus receives an extensive cholinergic input from the medial septal nucleus that ramifies throughout all layers and plays a pivotal modulatory role in cognitive function. Although the pharmacological effects of exogenous application of cholinergic agonists have been extensively studied in hippocampal neurons, much less is known about the effects of synaptically released acetylcholine (ACh). In this respect, most studies have focused on the cholinergic afferent input to pyramidal neurons that produces a characteristically slow depolarizing synaptic response mediated by activation of muscarinic ACh receptors (mAChRs). Here we report that cholinergic afferent stimulation also elicits atropine-sensitive synaptic potentials in hippocampal CA1 interneurons but, in contrast to synaptic responses in pyramidal neurons, these are highly diverse in waveform, although can still be classified into five distinct subtypes. The most common response type (i) 64% of cells) consisted of a slow sustained membrane potential depolarization. The other 36% of responses could be subdivided into responses comprising of (ii) a biphasic membrane potential change in which an initial slow hyperpolarization subsequently transforms into a slow depolarization (20%), (iii) a pure, slow hyperpolarization (13%), and (iv) an oscillatory response persisting for several seconds (2%). Interestingly, there were also interneurons totally insensitive to both synaptic and pharmacological cholinergic challenge. Morphological investigation of recorded cells revealed no obvious correlation between responsiveness to cholinergic afferent stimulation and dendritic and axonal arborization. The current study suggests that synaptic release of ACh results in a complex and differential mAChR-mediated modulation of cellular excitability within the hippocampal interneuron population.
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Potenciales Postsinápticos Excitadores/fisiología , Hipocampo/citología , Hipocampo/fisiología , Interneuronas/fisiología , Receptores Muscarínicos/fisiología , Acetilcolina/fisiología , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/fisiología , Animales , Atropina/farmacología , Carbacol/farmacología , Forma de la Célula/fisiología , Agonistas Colinérgicos/farmacología , Inhibidores de la Colinesterasa/farmacología , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Femenino , Interneuronas/efectos de los fármacos , Masculino , Antagonistas Muscarínicos/farmacología , Vías Nerviosas , Técnicas de Cultivo de Órganos , Fisostigmina/farmacología , Ratas , Ratas Wistar , Núcleos Septales/citología , Núcleos Septales/fisiología , Ácido gamma-Aminobutírico/fisiologíaRESUMEN
The level of arousal in mammals is correlated with metabolic state and specific patterns of cortical neuronal responsivity. In particular, rhythmic transitions between periods of high activity (up phases) and low activity (down phases) vary between wakefulness and deep sleep/anesthesia. Current opinion about changes in cortical response state between sleep and wakefulness is split between neuronal network-mediated mechanisms and neuronal metabolism-related mechanisms. Here, we demonstrate that slow oscillations in network state are a consequence of interactions between both mechanisms. Specifically, recurrent networks of excitatory neurons, whose membrane potential is partly governed by ATP-modulated potassium (K(ATP)) channels, mediate response-state oscillations via the interaction between excitatory network activity involving slow, kainate receptor-mediated events and the resulting activation of ATP-dependent homeostatic mechanisms. These findings suggest that K(ATP) channels function as an interface between neuronal metabolic state and network responsivity in mammalian cortex.
Asunto(s)
Corteza Cerebral/fisiología , Red Nerviosa , Neuronas/metabolismo , Adenosina Trifosfato/metabolismo , Animales , Corteza Cerebral/citología , Corteza Cerebral/metabolismo , Canales de Potasio/metabolismo , RatasRESUMEN
OBJECTIVE: To verify whether a deficiency in the cardiac sarcoplasmic reticulum pump SERCA2a causes cardiac dysfunction in humans. DESIGN: Cardiac performance was measured in a serendipitous human model of primary SERCA2a deficiency, Darier's disease, an autosomal dominant skin disorder caused by mutations inactivating one copy of the ATP2A2 gene, which encodes SERCA2a. METHODS: Systolic and diastolic function and contractility were assessed by echocardiography at rest and during exercise in patients with Darier's disease with known mutations. Fourteen patients with Darier's disease were compared with 14 normal controls and six patients with dilated cardiomyopathy with stable heart failure. RESULTS: Resting systolic and diastolic function was normal in patients with Darier's disease and in controls. The increase in systolic function during exercise was not different between patients with Darier's disease and normal controls; neither was there a difference in contractility. As expected, patients with dilated cardiomyopathy had impaired diastolic and systolic function with depressed contractility at rest and during exercise. CONCLUSION: Contrary to expectations, heterozygous disruption of SERCA2a is not associated with the impairment of cardiac performance in humans. Attempts to increase SERCA2a levels in heart failure, although showing promise in rodent studies, may not be addressing a critical causal pathway in humans.
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ATPasas Transportadoras de Calcio/genética , Cardiomiopatía Dilatada/genética , Enfermedad de Darier/genética , Insuficiencia Cardíaca/genética , Heterocigoto , ATPasas Transportadoras de Calcio/deficiencia , Estudios de Casos y Controles , Tolerancia al Ejercicio , Femenino , Terapia Genética/métodos , Insuficiencia Cardíaca/terapia , Humanos , Masculino , Persona de Mediana Edad , Mutación/genética , ATPasas Transportadoras de Calcio del Retículo SarcoplásmicoRESUMEN
Vanilloid receptor-1 (TRPV1) is a non-selective cation channel, predominantly expressed by peripheral sensory neurones, which is known to play a key role in the detection of noxious painful stimuli, such as capsaicin, acid and heat. To date, a number of antagonists have been used to study the physiological role of TRPV1; however, antagonists such as capsazepine are somewhat compromised by non-selective actions at other receptors and apparent modality-specific properties. SB-366791 is a novel, potent, and selective, cinnamide TRPV1 antagonist isolated via high-throughput screening of a large chemical library. In a FLIPR-based Ca(2+)-assay, SB-366791 produced a concentration-dependent inhibition of the response to capsaicin with an apparent pK(b) of 7.74 +/- 0.08. Schild analysis indicated a competitive mechanism of action with a pA2 of 7.71. In electrophysiological experiments, SB-366791 was demonstrated to be an effective antagonist of hTRPV1 when activated by different modalities, such as capsaicin, acid or noxious heat (50 degrees C). Unlike capsazepine, SB-366791 was also an effective antagonist vs. the acid-mediated activation of rTRPV1. With the aim of defining a useful tool compound, we also profiled SB-366791 in a wide range of selectivity assays. SB-366791 had a good selectivity profile exhibiting little or no effect in a panel of 47 binding assays (containing a wide range of G-protein-coupled receptors and ion channels) and a number of electrophysiological assays including hippocampal synaptic transmission and action potential firing of locus coeruleus or dorsal raphe neurones. Furthermore, unlike capsazepine, SB-366791 had no effect on either the hyperpolarisation-activated current (I(h)) or Voltage-gated Ca(2+)-channels (VGCC) in cultured rodent sensory neurones. In summary, SB-366791 is a new TRPV1 antagonist with high potency and an improved selectivity profile with respect to other commonly used TRPV1 antagonists. SB-366791 may therefore prove to be a useful tool to further study the biology of TRPV1.
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Anilidas/farmacología , Capsaicina/análogos & derivados , Cinamatos/farmacología , Péptidos y Proteínas de Señalización Intracelular , Potenciales de la Membrana/efectos de los fármacos , Receptores de Droga/antagonistas & inhibidores , 8-Hidroxi-2-(di-n-propilamino)tetralin/farmacología , Ácidos/farmacología , Anilidas/química , Compuestos de Anilina/metabolismo , Animales , Calcio/metabolismo , Capsaicina/farmacología , Proteínas Portadoras/farmacología , Línea Celular , Cinamatos/química , Relación Dosis-Respuesta a Droga , Interacciones Farmacológicas , Embrión de Mamíferos , Agonistas de Aminoácidos Excitadores/farmacología , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Calor , Humanos , Riñón , N-Metilaspartato/farmacología , Neuropéptidos/farmacología , Norepinefrina/farmacología , Orexinas , Técnicas de Placa-Clamp/métodos , Unión Proteica/efectos de los fármacos , Ensayo de Unión Radioligante/métodos , Ratas , Receptores de Droga/química , Agonistas de Receptores de Serotonina/farmacología , Xantenos/metabolismo , Ácido alfa-Amino-3-hidroxi-5-metil-4-isoxazol Propiónico/farmacologíaRESUMEN
Several psychiatric diseases, including schizophrenia, are thought to have a developmental aetiology, but to date no clear link has been made between psychiatric disease and a specific developmental process. LPA(1) is a G(i)-coupled seven transmembrane receptor with high affinity for lysophosphatidic acid. Although LPA(1) is expressed in several peripheral tissues, in the nervous system it shows relatively restricted temporal expression to neuroepithelia during CNS development and to myelinating glia in the adult. We report the detailed neurological and behavioural analysis of mice homozygous for a targeted deletion at the lpa(1) locus. Our observations reveal a marked deficit in prepulse inhibition, widespread changes in the levels and turnover of the neurotransmitter 5-HT, a brain region-specific alteration in levels of amino acids, and a craniofacial dysmorphism in these mice. We suggest that the loss of LPA(1) receptor generates defects resembling those found in psychiatric disease.
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Trastornos Mentales/genética , Trastornos Mentales/metabolismo , Fenotipo , Receptores Acoplados a Proteínas G/deficiencia , Animales , Encéfalo/metabolismo , Femenino , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Tiempo de Reacción/fisiología , Receptores Acoplados a Proteínas G/genética , Receptores del Ácido Lisofosfatídico , Reflejo de Sobresalto/fisiologíaRESUMEN
Nicotinic acetylcholine receptors (nAChRs) regulate neuronal excitability within the CNS. To assess the possible modulatory influence of nAChRs on epileptiform activity, a range of nAChR ligands were applied during experimentally induced epileptiform activity in rat hippocampal slices. Bath application of the potassium channel blocker 4-aminopyridine (4AP; 10-50 microM) resulted in the development of spontaneous epileptiform bursting activity in area CA3 that consisted of short duration (257+/-15 ms) field events occurring regularly at a frequency of 0.4+/-0.02 Hz. Subsequent co-application of the selective nAChR agonists 1,1-dimethyl-4-phenyl-piperazinium iodide (DMPP; 0.3-300 microM), choline (0.01-3mM) and lobeline (3-30 microM) produced sustained and concentration-dependent increases in burst frequency with maximal frequency potentiation of 37+/-5%, 27+/-5% and 24+/-11%, respectively. DMPP (10-30 microM; n=31) also potentiated epileptiform bursting induced by reducing GABA(A) receptor-mediated synaptic transmission using 20 microM bicuculline or enhancing NMDA receptor-mediated excitation by lowering extracellular Mg(2+). Irrespective of the epileptiform model studied all nAChR agonist induced frequency potentiation was reversed upon washout of the agonist or co-application of one of the selective nAChR antagonists dihydro-beta-erythroidine (10-30 microM), mecamylamine (50-200 microM) or alpha-bungarotoxin (100 nM). These results provide compelling evidence that activation of nAChRs exacerbate epileptiform activity in the rat hippocampus.
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Potenciales de Acción/fisiología , Epilepsia/fisiopatología , Hipocampo/fisiopatología , Receptores Nicotínicos/fisiología , 2-Amino-5-fosfonovalerato/farmacología , 4-Aminopiridina , Potenciales de Acción/efectos de los fármacos , Análisis de Varianza , Animales , Bicuculina , Colina/farmacología , Relación Dosis-Respuesta a Droga , Interacciones Farmacológicas , Electrofisiología/métodos , Epilepsia/inducido químicamente , Antagonistas de Aminoácidos Excitadores/farmacología , Hipocampo/efectos de los fármacos , Técnicas In Vitro , Masculino , Agonistas Nicotínicos/farmacología , Antagonistas Nicotínicos/farmacología , Ratas , Ratas Wistar , Análisis de RegresiónRESUMEN
GABA(B) receptor subunits are widely expressed on neurons throughout the CNS, at both pre- and postsynaptic sites, where they mediate the late, slow component of the inhibitory response to the major inhibitory neurotransmitter GABA. The existence of functional GABA(B) receptors on nonneuronal cells has been reported previously, although the molecular composition of these receptors has not yet been described. Here we demonstrate for the first time, using immunohistochemistry the expression of GABA(B1a), GABA(B1b), and GABA(B2) on nonneuronal cells of the rat CNS. All three principle GABA(B) receptor subunits were expressed on these cells irrespective of whether they had been cultured or found within brain tissue sections. At the ultrastructural level GABA(B) receptor subunits were expressed on astrocytic processes surrounding both symmetrical and assymetrical synapses in the CA1 subregion of the hippocampus. In addition, GABA(B1a), GABA(B1b), and GABA(B2) receptor subunits were expressed on activated microglia in culture but were not found on myelin forming oligodendrocytes in the white matter of rat spinal cord. Together these data demonstrate that the obligate subunits of functional GABA(B) receptors are expressed in astrocytes and microglia in the rat CNS.
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Astrocitos/metabolismo , Encéfalo/metabolismo , Microglía/metabolismo , Receptores de GABA-B/biosíntesis , Animales , Astrocitos/ultraestructura , Encéfalo/ultraestructura , Membrana Celular/metabolismo , Membrana Celular/ultraestructura , Células Cultivadas , Inmunohistoquímica , Masculino , Microglía/ultraestructura , Microscopía Electrónica , Terminales Presinápticos/metabolismo , Terminales Presinápticos/ultraestructura , Subunidades de Proteína/biosíntesis , Ratas , Ratas Sprague-Dawley , Transmisión Sináptica/fisiología , Ácido gamma-Aminobutírico/metabolismoRESUMEN
Neuronal networks of the hippocampal CA3 region generate stereotyped patterns of electrical activity in response to activation of metabotropic glutamate receptors (mGluRs) or muscarinic acetylcholine receptors (mAChRs) that consist of intermittent episodes of prolonged oscillatory activity. In light of the slow kinetics of such network responses, we investigated the possible contribution of the hyperpolarisation-activated inward current (I(h)) in the generation and maintenance of hippocampal oscillatory states. Hippocampal 'mini-slice' experiments in which the main subfields of the hippocampus were isolated by transection of the connecting afferents revealed that the CA3 region was the primary generator of both mGluR and mAChR-mediated network responses. Subsequent patch-clamp experiments confirmed the presence of a prominent hyperpolarisation-activated inward current in the principal cells of the CA3 region that was sensitive to caesium chloride and the selective I(h) blocker ZD-7288.Furthermore, in the presence of mAChR or mGluR agonists these cells exhibited a slow membrane potential oscillation that was independent of AMPA receptor-mediated synaptic transmission. Blockade of I(h) suppressed this oscillation as well as mGluR and mAChR-induced theta based intermittent network oscillatory behaviour. These data support the idea that the I(h) pacemaker current is important in the generation of patterned neuronal activities in the hippocampus.
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Hipocampo/fisiología , Red Nerviosa/fisiología , Receptores de Glutamato Metabotrópico/fisiología , Receptores Muscarínicos/fisiología , Ritmo Teta , Animales , Carbacol/farmacología , Cardiotónicos/farmacología , Agonistas Colinérgicos/farmacología , Cicloleucina/farmacología , Agonistas de Aminoácidos Excitadores/farmacología , Antagonistas de Aminoácidos Excitadores/farmacología , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Potenciales Postsinápticos Excitadores/fisiología , Femenino , Hipocampo/anatomía & histología , Hipocampo/citología , Hipocampo/efectos de los fármacos , Técnicas In Vitro , Técnicas de Placa-Clamp/métodos , Células Piramidales/efectos de los fármacos , Células Piramidales/fisiología , Pirimidinas/farmacología , Quinoxalinas/farmacología , Ratas , Ratas Wistar , Receptores de Glutamato Metabotrópico/efectos de los fármacos , Receptores Muscarínicos/efectos de los fármacos , Transmisión Sináptica/efectos de los fármacos , Ritmo Teta/efectos de los fármacosRESUMEN
AIMS: To assess the heritability (i.e. relative contribution of genetic factors to the variability) of continuous measures of left ventricular hypertrophy determined by electrocardiography and echocardiography. METHODS AND RESULTS: We studied 955 members of 229 Caucasian families, ascertained through a hypertensive proband. Electrocardiographic measurements were performed manually on resting 12-lead electrocardiograms, and echocardiographic measurements were made on M-mode images. Sex-specific residuals for the left ventricular phenotypes were calculated, adjusted for age, systolic blood pressure, weight, height, waist-hip ratio, and presence of diabetes. Heritability was estimated in two ways: firstly, from familial correlations with adjustment for spouse resemblance; and secondly by using variance components methods with ascertainment correction for proband status. The heritability estimates (given as a range derived from the two methods) were higher for Sokolow-Lyon voltage (39-41%) than for echocardiographic left ventricular mass (23-29%). Electrocardiographic left ventricular mass, Cornell voltage, and Cornell product had heritability estimates of 12-18%, 19-25%, and 28-32%, respectively. CONCLUSIONS: Genetic factors may explain a substantial proportion of variability in quantitative electrocardiographic and echocardiographic measures of left ventricular hypertrophy. The greater heritability of Sokolow-Lyon voltage suggests that electrocardiographic phenotypes may be particularly important for the molecular investigation of the genetic susceptibility to cardiac hypertrophy.
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Hipertrofia Ventricular Izquierda/genética , Ecocardiografía/métodos , Electrocardiografía/métodos , Composición Familiar , Femenino , Humanos , Hipertrofia Ventricular Izquierda/patología , Masculino , Persona de Mediana Edad , Linaje , FenotipoRESUMEN
Theta frequency oscillations are a predominant feature of rhythmic activity in the hippocampus. We demonstrate that hippocampal area CA1 generates atropine-resistant theta population oscillations in response to metabotropic glutamate receptor activation under conditions of reduced AMPA receptor activation. This activity occurred in the absence of inputs from area CA3 and extra-ammonic areas. Field theta oscillations were co-expressed with pyramidal distal apical dendritic burst spiking and were temporally related to trains of IPSPs with slow kinetics. Pyramidal somatic responses showed theta oscillations consisted of compound inhibitory synaptic potentials with initial IPSPs with slow kinetics followed by trains of smaller, faster IPSPs. Pharmacological modulation of IPSPs altered the theta oscillation suggesting an inhibitory network origin. Somatic IPSPs, dendritic burst firing and stratum pyramidale interneuron activity were all temporally correlated with spiking in stratum oriens interneurons demonstrating intrinsic theta-frequency oscillations. Disruption of spiking in these interneurons was accompanied by a loss of both field theta and theta frequency IPSP trains. We suggest that population theta oscillations can be generated as a consequence of intrinsic theta frequency spiking activity in a subset of stratum oriens interneurons controlling electrogenesis in pyramidal cell apical dendrites.
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Atropina/farmacología , Hipocampo/fisiología , Parasimpatolíticos/farmacología , Ritmo Teta/efectos de los fármacos , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/fisiología , Animales , Dendritas/fisiología , Antagonistas de Aminoácidos Excitadores/farmacología , Hipocampo/citología , Hipocampo/efectos de los fármacos , Interneuronas/fisiología , Interneuronas/ultraestructura , Masculino , Inhibición Neural/fisiología , Quinoxalinas/farmacología , Ratas , Ratas Wistar , Receptores AMPA/antagonistas & inhibidores , Receptores AMPA/fisiología , Receptores de Glutamato Metabotrópico/fisiologíaRESUMEN
Learning and memory involve complex changes in neuronal excitability including long-lasting synaptic plasticity of glutamatergic synapses. The observation that the neuropeptide galanin affects performance in a number of behavioural models predicts that galanin should affect synaptic processes underlying learning. The present study in rat and mouse hippocampal slices now demonstrates that galanin inhibits long-term potentiation induced by both tetanic and theta-burst stimulation in both apical and basal dendrites of CA1 pyramidal neurones but does not affect long-term depression. This selective effect on long-term potentiation does not appear to be mediated through inhibition of N-methyl-D-aspartate or metabotropic glutamate receptor function, but likely resides downstream of receptor activation, possibly at the level of the kinase cascade that converts short-term into long-term potentiation. Our results indicate possible mechanisms by which the neuropeptide galanin may act at the molecular level to influence learning and memory in vertebrates.
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Galanina/farmacología , Hipocampo/efectos de los fármacos , Hipocampo/fisiología , Plasticidad Neuronal/efectos de los fármacos , Sinapsis/fisiología , Animales , Estimulación Eléctrica , Femenino , Técnicas In Vitro , Potenciación a Largo Plazo/efectos de los fármacos , Potenciación a Largo Plazo/fisiología , Ratones , Ratones Endogámicos , Ratas , Ratas Wistar , Sinapsis/efectos de los fármacos , Ritmo Teta , Factores de TiempoRESUMEN
Electrophysiological recordings from identified noradrenergic locus coeruleus (LC) neurones in rat brain slices have revealed that the orexins can cause direct and reversible depolarisation of the postsynaptic membrane. Whilst it is known that the membrane depolarisation produced by orexin-A can triple the firing rate of spontaneously active LC neurones, quantitative pharmacological analysis that determines the receptor subtype(s) mediating the orexinergic response has not yet been performed. Here we demonstrate that the effects of orexin-A are five-fold more potent than orexin-B on LC neuronal excitability. We show further that the orexin receptor antagonist SB-334867-A inhibits the effects of both agonists with pK(B) values similar to those calculated for human OX1 receptors expressed in CHO cells. Finally, we found no evidence for tonic activation of OX1 receptors in LC noradrenergic neurones despite electron microscopic evidence that orexin terminals directly contact these neurones. These data demonstrate that SB-334867-A is a useful tool compound with which to study the physiology of OX1 receptors.
Asunto(s)
Benzoxazoles/farmacología , Proteínas Portadoras/antagonistas & inhibidores , Péptidos y Proteínas de Señalización Intracelular , Locus Coeruleus/efectos de los fármacos , Neuropéptidos/antagonistas & inhibidores , Urea/farmacología , Potenciales de Acción/efectos de los fármacos , Animales , Proteínas Portadoras/farmacología , Electrofisiología , Técnicas In Vitro , Masculino , Naftiridinas , Neuropéptidos/farmacología , Receptores de Orexina , Orexinas , Técnicas de Placa-Clamp , Ratas , Ratas Sprague-Dawley , Receptores Acoplados a Proteínas G , Receptores de Glutamato Metabotrópico/efectos de los fármacos , Receptores de Glutamato Metabotrópico/metabolismo , Receptores Muscarínicos/efectos de los fármacos , Receptores Muscarínicos/metabolismo , Receptores de Neuropéptido , Urea/análogos & derivadosRESUMEN
The aim of this review is firstly to describe the current understanding of the diverse physiology and pharmacology of GABA(B) receptors in vivo. We will then focus on recent advances made, since the identification of the GABA(B) receptor subunit genes, in our knowledge of the molecular nature of the receptor, and the recently discovered molecular determinants of functions such as ligand binding, trafficking and signalling. We will conclude with a summary of the GABA(B) receptor-interacting proteins that have been described thus far, and discuss how these may, at least in part, account for the paradox of varied receptor pharmacology in the potential context of a single heterodimeric GABA(B) receptor.
Asunto(s)
Receptores de GABA-B/deficiencia , Sinapsis/metabolismo , Transmisión Sináptica/genética , Animales , Humanos , Ligandos , Inhibición Neural/efectos de los fármacos , Inhibición Neural/genética , Estructura Terciaria de Proteína/efectos de los fármacos , Estructura Terciaria de Proteína/genética , Subunidades de Proteína/efectos de los fármacos , Subunidades de Proteína/genética , Subunidades de Proteína/metabolismo , Receptores de GABA-B/efectos de los fármacos , Receptores de GABA-B/genética , Sinapsis/efectos de los fármacos , Transmisión Sináptica/efectos de los fármacosRESUMEN
Recent experiments have demonstrated that formation of functional type B gamma-aminobutyric acid (GABA(B)) receptors requires co-expression of two receptor subunits, GABA(B1) and GABA(B2). Despite the identification of these subunits and a number of associated splice variants, there has been little convincing evidence of pharmacological diversity between GABA(B) receptors comprising different subunit combinations. However, Ng et al. [Mol. Pharmacol., 59 (2000) 144] have recently suggested a novel and important pharmacological difference between GABA(B) receptor heterodimers expressing the GABA(B1a) and GABA(B1b) receptor subunits. This study suggested that the antiepileptic GABA analogue gabapentin (Neurontin) is an agonist at GABA(B) receptors expressing the GABA(B1a) but not the GABA(B1b) receptor subunit. The importance of this finding with respect to identifying novel GABA(B) receptor subunit specific agonists prompted us to repeat these experiments in our own [35S]-GTPgammaS binding and second messenger assay systems. Here we report that gabapentin was completely inactive at recombinant GABA(B) heterodimers expressing either GABA(B1a) or GABA(B1b) receptor subunits in combination with GABA(B2) receptor subunits. In addition, in both CA1 and CA3 pyramidal neurones from rodent hippocampal slices we were unable to demonstrate any agonist-like effects of gabapentin at either pre- or post-synaptic GABA(B) receptors. In contrast, gabapentin activated a GABA(A) receptor mediated chloride conductance. Our data suggest that gabapentin is not a GABA(B)-receptor agonist let alone a GABA(B) receptor subunit selective agonist.
Asunto(s)
Acetatos/farmacología , Aminas , Anticonvulsivantes/farmacología , Ácidos Ciclohexanocarboxílicos , Agonistas del GABA/farmacología , Agonistas de Receptores GABA-B , Ácido gamma-Aminobutírico , Acetatos/metabolismo , Secuencia de Aminoácidos , Animales , Anticonvulsivantes/metabolismo , Unión Competitiva , Relación Dosis-Respuesta a Droga , Agonistas del GABA/metabolismo , Antagonistas del GABA/metabolismo , Antagonistas del GABA/farmacología , Antagonistas de Receptores de GABA-B , Gabapentina , Hipocampo/efectos de los fármacos , Hipocampo/metabolismo , Ratones , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Ratas , Receptores de GABA-B/metabolismoRESUMEN
OBJECTIVE: To evaluate the accuracy of a three-dimensional (3D) magnetic position sensor system in the quantification of ventricular stroke volumes in a dynamic model. METHODS: A latex balloon model of the left ventricle was suspended in a water bath connected to a pump producing 10 different pulsatile stroke volumes (15-65 ml/beat). Scanning was performed using a 5.0 mHz transducer mounted with a Flock of Birds (FOB) magnetic receiver (GE System Five). The probe was scanned to sweep continuously across and over the balloon volume over 3-7 seconds. Digital loops were stored on magneto-optical disks and reviewed retrospectively using 3D Echopac software (GE) based on Simpson's method and compared with a two-dimensional (2D) biplane area-length method (1/2 L x R) measurements at end systole and end diastole. Both 3D and 2D derived stroke volumes were compared with the reference stroke volume calculated by direct measurement of balloon capacity. RESULTS: There was an improved correlation between 3D stroke volume and reference stroke volume (y = 0.91x + 0.41, r = 0.97, SEE = 2.83 ml, P = 0.0001) compared to 2D stroke volume and reference stroke volume (y = 0.49x + 8.68, r = 0.87, SEE = 3.87 ml, P = 0.0011, difference between 2D and 3D P < 0.003). CONCLUSION: 3D magnetic FOB scanning is practical, accurate and should facilitate assessment of left ventricular function.