RESUMEN
The loss of dopamine in idiopathic or animal models of Parkinson's disease induces synchronized low-frequency oscillatory burst-firing in subthalamic nucleus neurones. We sought to establish whether these firing patterns observed in vivo were preserved in slices taken from dopamine-depleted animals, thus establishing a role for the isolated subthalamic-globus pallidus complex in generating the pathological activity. Mice treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) showed significant reductions of over 90% in levels of dopamine as measured in striatum by high pressure liquid chromatography. Likewise, significant reductions in tyrosine hydroxylase immunostaining within the striatum (>90%) and tyrosine hydroxylase positive cell numbers (65%) in substantia nigra were observed. Compared with slices from intact mice, neurones in slices from MPTP-lesioned mice fired significantly more slowly (mean rate of 4.2 Hz, cf. 7.2 Hz in control) and more irregularly (mean coefficient of variation of inter-spike interval of 94.4%, cf. 37.9% in control). Application of ionotropic glutamate receptor antagonists 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 2-amino-5-phosphonopentanoic acid (AP5) and the GABA(A) receptor antagonist picrotoxin caused no change in firing pattern. Bath application of dopamine significantly increased cell firing rate and regularized the pattern of activity in cells from slices from both MPTP-treated and control animals. Although the absolute change was more modest in control slices, the maximum dopamine effect in the two groups was comparable. Indeed, when taking into account the basal firing rate, no differences in the sensitivity to dopamine were observed between these two cohorts. Furthermore, pairs of subthalamic nucleus cells showed no correlated activity in slices from either control (21 pairs) or MPTP-treated animals (20 pairs). These results indicate that the isolated but interconnected subthalamic-globus pallidus network is not itself sufficient to generate the aberrant firing patterns in dopamine-depleted animals. More likely, inputs from other regions, such as the cortex, are needed to generate pathological oscillatory activity.
Asunto(s)
1-Metil-4-fenil-1,2,3,6-Tetrahidropiridina/farmacología , Potenciales de Acción/efectos de los fármacos , Dopamina/metabolismo , Neuronas/efectos de los fármacos , Neurotoxinas/farmacocinética , Núcleo Subtalámico/citología , Animales , Química Encefálica/efectos de los fármacos , Cromatografía Líquida de Alta Presión/métodos , Dopamina/farmacología , Relación Dosis-Respuesta a Droga , Interacciones Farmacológicas , Inmunohistoquímica/métodos , Técnicas In Vitro , Masculino , Ratones , Ratones Endogámicos C57BL , Neuronas/metabolismo , Núcleo Subtalámico/efectos de los fármacos , Núcleo Subtalámico/metabolismoRESUMEN
Dopamine, acting at a D1-like receptor, depresses the release of glutamate in the nucleus accumbens (NAcc) in brain slices, thereby reducing the amplitude of the excitatory postsynaptic current (EPSC). This effect depends upon an inhibitory feedback action of adenosine, liberated following facilitation of postsynaptic NMDA receptors by D1 receptor activation, an action independent of adenylyl cyclase stimulation or cyclic AMP-dependent protein kinase (PKA; Harvey, J., Lacey, M.G., 1997. J. Neurosci. 17, 5271). Using whole-cell recording from NAcc neurones, the dopamine depression of the EPSC was blocked by pre-treatment of brain slices with the selective protein kinase C (PKC) inhibitor Ro 32-0432, but only minimally attenuated by intracellular dialysis of single cells with Ro 32-0432 in the recording pipette. With synaptic transmission blocked by tetrodotoxin, inward currents caused by application of NMDA were enhanced by the D1 receptor agonist SKF 81297A in half the cells tested. In a separate population of cells dialysed intracellularly with Ro 32-0432, SKF 81297A was without effect on NMDA current amplitude. These findings indicate a functional role for phospholipase C-coupled D1-like receptors in both modulating synaptic transmission in NAcc and potentiating NMDA receptors on a subset of NAcc neurones, via PKC activation.
Asunto(s)
Inhibidores Enzimáticos/farmacología , Indoles/farmacología , Neuronas/fisiología , Núcleo Accumbens/fisiología , Pirroles/farmacología , Receptores de Dopamina D1/fisiología , Receptores de N-Metil-D-Aspartato/fisiología , Sinapsis/fisiología , Transmisión Sináptica/fisiología , Animales , Dopamina/farmacología , Potenciales Evocados/efectos de los fármacos , Potenciales Evocados/fisiología , Técnicas In Vitro , Masculino , Potenciales de la Membrana/efectos de los fármacos , Potenciales de la Membrana/fisiología , Neuronas/efectos de los fármacos , Núcleo Accumbens/efectos de los fármacos , Técnicas de Placa-Clamp , Proteína Quinasa C/antagonistas & inhibidores , Ratas , Ratas Wistar , Receptores de Dopamina D1/efectos de los fármacos , Receptores de N-Metil-D-Aspartato/efectos de los fármacos , Sinapsis/efectos de los fármacos , Transmisión Sináptica/efectos de los fármacosRESUMEN
Intracellular recordings were made from neurones in the rat locus coeruleus in a brain slice maintained in vitro. Phencyclidine and related psychotomimetic drugs, applied in known concentrations in the fluid bathing the slice, depressed responses to N-methyl-D-aspartic acid noradrenaline (in the presence of the uptake inhibitor desmethylimipramine) and [D-Ala2,MePhe4,Gly-ol5]enkephalin and also prolonged the action potential. The sensitivities of these responses to depression by phencyclidine was N-methyl-D-aspartic acid (IC50 0.4 microM) greater than noradrenaline (IC50 3.9 microM) greater than [D-Ala2,MePhe4,Gly-ol5]enkephalin (IC50 8.5 microM) greater than prolongation of the action potential (41% increase by 30 microM). Stereoselectivity was observed only in the depression of responses to N-methyl-D-aspartic acid where (+)-1-(1-phenylcyclohexyl)-3-methyl piperidine was 3.3-fold more potent in suppressing N-methyl-D-aspartic acid depolarizations than its (-) isomer. The responses to N-methyl-D-aspartic acid were also depressed by the structurally unrelated psychotomimetic (+/-)-N-allyl-N-normetazocine (IC50 0.9 microM). All of the effects of the psychotomimetic drugs examined were slow in onset and difficult to reverse following washout. No effect of phencyclidine (0.03-100 microM) or related drugs was observed on membrane potential, input resistance or spontaneous action potential firing rate of locus coeruleus neurones. The depression of responses to N-methyl-D-aspartic acid by phencyclidine was the most potent and the only stereoselective effect of those studied. The importance of this effect and of those not showing stereoselectivity in relation to the phencyclidine behavioural syndrome is discussed.
Asunto(s)
Locus Coeruleus/efectos de los fármacos , Neuronas/efectos de los fármacos , Fenciclidina/farmacología , Potenciales de Acción/efectos de los fármacos , Animales , Ácido Aspártico/análogos & derivados , Ácido Aspártico/farmacología , Recuento de Células , Relación Dosis-Respuesta a Droga , Encefalina Ala(2)-MeFe(4)-Gli(5) , Encefalinas/farmacología , Potenciales Evocados/efectos de los fármacos , Locus Coeruleus/fisiología , Masculino , Potenciales de la Membrana/efectos de los fármacos , N-Metilaspartato , Neuronas/fisiología , Norepinefrina/farmacología , Fenciclidina/análogos & derivados , Ratas , Ratas Endogámicas , Estereoisomerismo , Factores de TiempoRESUMEN
Adenosine trisphosphate-sensitive potassium (K-ATP) channels in the substantia nigra pars reticulata were studied in rat brain slices using whole-cell patch clamp recording. Substantia nigra pars reticula neurons were identified as such by their spontaneous action potential firing at mean rate of 15.3 Hz1 virtual absence of hyperpolarization-activated inward current Ih1 and unresponsiveness to dopamine (30 microM), quinirole (10 microM) and (Met)enkephalin (10 microM). Intracellular dialysis with Mg(2+0-ATP-free pipette solutions caused a slowly developing membrane hyperpolarization (13 +/- 4 mV), accompanied by a cessation of action potential firing, or an outward current (79 +/- 30 pA at around -60 mV), which were reversed b the sulphonylurea K-ATO channel blockers tolbutamide (100 microM) and glibenclamide (3 microM). When Mg(2+0-ATP (2 mM) was included in the recording pipette no membrane hyperpolarization or outward current was observed. Neither the sulphonylureas nor the potassium channel activator lemakalim (200 MicroM) altered membrane potential, firing rate or holding current under these recording conditions. The outward current induced by dialysis with Mg(2+)-ATP-free solutions reversed polarity negative to -94 +/- 9 mV (9 cells), close to the estimated K+ equilibrium potential (-105 mV) for the conditions used, and was associated with a conductance increase that was blocked by Ba2+ (100 microM). The current blocked by the sulphonylureas had a similar reversal potential (-97 +/- 7 MV; 13 cells), and both currents were voltage independent over the range -50 to -100 mV with slope conductance of approximately 2.0 nS. Outward synaptic current were evoked by single shock electrical simulation, in the presence of glutamate receptor antagonists, at a holding potential of -50 mV. These synaptic currents were blocked by bicuculline (10 microM) and reversed polarity at around -65 mV, close to the Cl- equilibrium potential, and were thus mediated by GABAA receptors. They were reversibly depressed by 37 +/- 14% in lemakalim (200 microM) in 6/12 cells tested, an effect that was partially reversed by tolbutamide (200 microM). It is concluded that functional K-ATP channels are present both presynaptically and postsynaptically in the substantia nigra pars reticulata. Postsynaptic K-ATP channels may control excitability in conditions where intracellular ATP is reduced, whereas presynaptic K-ATP channels, sensitive to the potassium channel activator lemakalim, can modulate the release of GABA, which probably arises from fibres of extranigral origin. Pharmacological differences between these two sites could be exploited to treat epilepsies, dyskinesias and akinesia.
Asunto(s)
Adenosina Trifosfato/farmacología , Dopamina/farmacología , Encefalinas/farmacología , Canales de Potasio/efectos de los fármacos , Quinpirol/farmacología , Sustancia Negra/efectos de los fármacos , Animales , Potenciales de la Membrana/efectos de los fármacos , Técnicas de Placa-Clamp , Ratas , Ratas Wistar , Tolbutamida/farmacologíaRESUMEN
In Parkinson's disease the neurones of the subthalamic nucleus show increased synchrony and oscillatory burst discharge, thought to reflect a breakdown of parallel processing in basal ganglia circuitry. To understand better the mechanisms underlying this transition, we sought to mimic this change in firing pattern within sagittal slices of rat midbrain. The firing patterns of up to four simultaneously extracellularly recorded subthalamic nucleus (STN) neurones were analysed using burst and oscillation detection programs, and correlated activity between pairs of neurones assessed. In control conditions all but 11 of 488 (2%) neurones fired in a predominantly tonic pattern (with mean oscillation frequency >3 Hz), with no significantly cross-correlated activity in any of 393 pairs of neurones. The glutamate antagonists DL-2-amino-phosphonopentanoic acid (APV), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and 6-methyl-2-(phenylethynyl)pyridine (MPEP) did not change the firing rate or pattern of these cells, providing no evidence for a role of glutamatergic collaterals within the STN under these conditions. The GABA(A) receptor antagonist bicuculline and GABA(B) receptor antagonist (2S)-3-[[(1S)-1-(3,4-dichlorophenyl)ethyl]amino-2-hydroxypropyl]phenylmethyl phosphinic acid (CGP 55845) were also without effect on firing rate or pattern in these cells, suggesting that there was no active input from other GABAergic basal ganglia nuclei in this slice. The dopamine receptor antagonist haloperidol caused no significant change to firing rate or pattern of firing in these cells, suggesting that there was no active dopaminergic input in this slice. Excitations of STN neurones by muscarine, (+)-1-aminocyclopentane-trans-1,3-dicarboxylic acid (ACPD), N-methyl-D-aspartic acid (NMDA) or dopamine were all unaccompanied by a change in firing pattern or any significant correlated activity between STN neurone pairs. Burst firing could be induced in STN neurones with either the potassium channel blocker tetraethylammonium (TEA; 10 mM; in 100/138 [72%] of cells) or with a combination of NMDA and the calcium-activated potassium channel blocker apamin (in 101/216 [47%] of cells). Burst firing in TEA was unchanged by CNOX and APV, MPEP, CGP55845, haloperidol, dopamine, and ACPD, although muscarine produced a significant increase in oscillation frequency. Burst firing in NMDA and apamin was unchanged by CNQX and APV, dopamine, muscarine and ACPD, although bicuculline caused a significant increase in oscillation frequency. Such burst firing was not accompanied by synchrony in any condition, either alone, or during application of excitatory agents or glutamate or GABA antagonists. As the bursting seen here was unaccompanied by the synchronous activity that has often been observed (pathologically) in vivo, it probably reflects solely intrinsic STN neuronal properties, rather than network activity. No functional role was found for glutamatergic collaterals within the STN, either when cells are firing tonically or burst firing. The circuitry needed to produce synchrony in the STN is most likely not intrinsic to the STN itself, but requires connections with other basal ganglia nuclei, and/or the cortex, which are not present in this preparation.
Asunto(s)
Potenciales de Acción/fisiología , Relojes Biológicos/fisiología , Neuronas/fisiología , Núcleo Subtalámico/fisiología , Potenciales de Acción/efectos de los fármacos , Animales , Relojes Biológicos/efectos de los fármacos , Encéfalo/efectos de los fármacos , Encéfalo/fisiología , Técnicas In Vitro , Masculino , N-Metilaspartato/farmacología , Red Nerviosa/efectos de los fármacos , Red Nerviosa/fisiología , Neuronas/efectos de los fármacos , Ratas , Ratas Wistar , Núcleo Subtalámico/efectos de los fármacos , Tetraetilamonio/farmacologíaRESUMEN
Intracellular recordings were made from neurons of the rat locus coeruleus contained within a brain slice maintained in vitro. When applied to the slice in known concentrations the selective kappa opioid receptor agonist trans-(+)-3,4-dichloro-N-methyl-[2-(1-pyrrolidinyl)cyclohexyl] benzeneacetamide methane sulphonate (U50488) (0.01-1 microM) produced a concentration-dependent depression of the excitatory post-synaptic potential evoked by electrical stimulation of afferent inputs to the locus coeruleus. This effect was antagonized by naloxone with an apparent dissociation equilibrium constant (Kd) of 28 nM. U50488 did not completely abolish the EPSP. Over the same concentration range U50488 had no effect on the resting membrane potential, input resistance or action potential waveform of locus coeruleus neurons, nor did U50488 depress the depolarization produced by local application of L-glutamic acid. The mu opioid receptor agonists [D-Ala2, NMe Phe4, Gly-ol5] enkephalin (0.003-1 microM) and [D-Ala2, NMe Phe4, Met(O)5] enkephalinol (0.003-1 microM) caused a membrane hyperpolarization concomitant with a fall in neuronal input resistance. These effects were concentration-dependent and antagonized by naloxone with an apparent Kd of 1.5 nM. Mu agonists also caused a depression of the tetrodotoxin resistant action potential. An in vitro autoradiographic study of [3H]bremazocine binding within the locus coeruleus revealed that, although the majority of binding appears to be to mu sites, a significant proportion was displaceable by unlabelled U50488 and thus represented kappa binding sites. The possibility that kappa opioid receptors may be located pre-synaptically within the locus coeruleus, and that activation of these receptors depresses excitatory synaptic input, is discussed.
Asunto(s)
Locus Coeruleus/fisiología , Pirrolidinas/farmacología , Receptores Opioides/fisiología , 3,4-Dicloro-N-metil-N-(2-(1-pirrolidinil)-ciclohexil)-bencenacetamida, (trans)-Isómero , Animales , Calcio/fisiología , Conductividad Eléctrica , Masculino , Potenciales de la Membrana/efectos de los fármacos , Naloxona/farmacología , Ratas , Ratas Endogámicas , Receptores Opioides/efectos de los fármacos , Receptores Opioides kappa , Sinapsis/fisiología , Transmisión SinápticaRESUMEN
Single-unit extracellular and whole-cell patch clamp recording were used to study the actions of exogenously applied 5-hydroxytryptamine on substantia nigra pars reticulata neurons in parasaggital slices of rat midbrain. Seventy-six per cent of substantia nigra pars reticulata cells (254/334) recorded extracellularly were excited by 5-hydroxytryptamine (EC50 = 9.56 microM); in the remainder, inhibitions (13.5%), biphasic responses (4.2%) or lack of response (6.3%) were observed. Using whole-cell patch recording, 5-hydroxytryptamine (10 microM) caused either an inward current (9/9 cells) or a depolarization (3/3 cells) at membrane potentials in the range -50 to -90 mV, which was resistant to tetrodotoxin (4/4 cells), indicating that the predominant, excitatory action of 5-hydroxytryptamine was due to a direct action on substantia nigra pars reticulata neurons. The 5-hydroxytryptamine excitation (recorded extracellularly) was reduced to 24 +/- 6% of control values by methysergide (0.1 microM) and to 17 +/- 5% of control by ketanserin (10 microM), but was unaffected by the 5-hydroxytryptamine antagonists spiperone (0.1 microM), yohimbine (0.1 microM), pindolol (1 microM), GR113808A (1 microM) or ICS 205930 (10 microM). In addition, the 5-hydroxytryptamine excitation was mimicked by the 5-hydroxytryptamine2C receptor--preferring agonist alpha-methyl 5-hydroxytryptamine (10 microM), but the agonists CP93, 129 (0.1-1 microM) and (+/-)-2-dipropylamino-8-hydroxy-1,2,3,4-tetrahydronaphthalene hydrobromide (0.1-1 microM) were without effect. Taken together, this pharmacology indicated involvement of the 5-hydroxytryptamine2C receptor in the 5-hydroxytryptamine excitation, while other candidate receptors known to be present in rat substantia nigra pars reticulata (5-hydroxytryptamine1B, 5-hydroxytryptamine2A and 5-hydroxytryptamine4) could be excluded from consideration. While in accord with current information on the location of 5-hydroxytryptamine receptor subtypes in substantia nigra pars reticulata, and the consequence of activation of neuronal 5-hydroxytryptamine2C receptors, these results contrast with data from in vivo experiments which suggest that the net effect of 5-hydroxytryptamine is to inhibit substantia nigra pars reticulata neurons. The reason for this apparent discrepancy may lie in detailed consideration of the microcircuitry of the substantia nigra pars reticulata. This may lead to a re-evaluation of the influence of 5-hydroxytryptamine on this basal ganglia output relay nucleus, and its role in motor control and the gating of generalized seizure activity.
Asunto(s)
Neuronas/efectos de los fármacos , Receptores de Serotonina/fisiología , Serotonina/farmacología , Sustancia Negra/efectos de los fármacos , Animales , Electrofisiología , Técnicas In Vitro , Ketanserina/farmacología , Masculino , Metisergida/farmacología , Ratas , Ratas Sprague-Dawley , Antagonistas de la Serotonina/farmacología , Agonistas de Receptores de Serotonina/farmacología , Sustancia Negra/citología , Sustancia Negra/fisiologíaRESUMEN
Recent anatomical and physiological studies have pointed to a functional innervation of the subthalamic nucleus by dopamine. This nucleus has a pivotal role in basal ganglia function and voluntary movement control and the possibility that dopamine, and dopaminergic medication used in Parkinson's disease, might directly influence its activity is of considerable interest. We have evaluated electrophysiologically the action and pharmacology of dopamine on single subthalamic neurones in rat brain slices. Dopamine increased firing rate to up to a mean of 60% in 98% of the 261 neurones tested when examined using extracellular single-unit recording. This excitation was unaffected by the GABA antagonist picrotoxin, and the glutamate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione, and persisted in a low Ca(2+)/raised Mg(2+) solution, indicative of a direct action, independent of synaptic transmission. Of the 33 cells examined using whole patch-clamp recording, only 13 showed measurable increases in firing rate and/or depolarisations in response to dopamine. Dopamine-responsive cells displayed significantly greater access resistance, suggesting that an unidentified cytoplamic constituent, removed by whole-cell dialysis, was required for the response. Using extracellular recording, the D2-like dopamine receptor agonists quinpirole and bromocryptine, but not the D1-like receptor agonist 1-phenyl-2,3,4,5-tetrahydro-(1H)-3-benzazepine-7,8-diol, also consistently caused an excitation. This was mimicked by the catecholamine releaser amphetamine in 60% of cells tested. However, the dopamine excitation was not significantly reduced either by the D1-like receptor antagonist 7-chloro8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine or the D2-like receptor antagonists (-)-sulpiride, eticlopride and (+)-butaclamol, and the quinpirole excitation was also unaffected by (-)-sulpiride. In contrast, (-)-sulpiride, eticlopride and (+)-butaclamol all abolished the D2-like receptor-mediated inhibition by dopamine of substantia nigra pars compacta neurones. The alpha-adrenoceptor antagonist phentolamine was a weak antagonist of dopamine excitations, but not of those caused by quinpirole. Dopamine excitations also showed weak sensitivity to the 5-HT(2) antagonist ritanserin, but were unaffected by the alpha(1)-adrenoceptor antagonist prazocin and the beta-adrenoceptor antagonist propranolol. The pharmacology of this dopamine excitation is inconsistent with an action on any known catecholamine receptor. However, the effect of amphetamine indicates that an unidentified monamine--possibly dopamine--can be released within the subthalamic nucleus to cause an excitation. The anomalies of its pharmacological characterisation do not strongly support a physiologically relevant direct action of dopamine in the rat subthalamic nucleus.
Asunto(s)
Agonistas de Dopamina/farmacología , Antagonistas de Dopamina/farmacología , Dopamina/fisiología , Neuronas/fisiología , Receptores Dopaminérgicos/fisiología , Núcleo Subtalámico/fisiología , Anfetamina/farmacología , Animales , Dopamina/farmacología , Inhibidores de Captación de Dopamina/farmacología , Relación Dosis-Respuesta a Droga , Electrofisiología , Masculino , Neuronas/efectos de los fármacos , Ratas , Ratas Wistar , Núcleo Subtalámico/efectos de los fármacosRESUMEN
1. Glutamate (AMPA receptor-mediated) excitatory postsynaptic potentials (e.p.s.ps.), evoked by electrical stimulation rostral to the recording site, were examined by intracellular microelectrode recording from dopamine neurones in parasagittal slices of rat ventral midbrain. 2. The e.p.s.p. was depressed by the group III metabotropic glutamate (mGlu) receptor agonist L-2-amino-4-phosphonobutyric acid (L-AP4; 0.01-30 microM) by up to 60% with an EC50 of 0.82 microM. The depression induced by L-AP4 (3 microM) was reversed by the group III preferring mGlu receptor antagonist, alpha-methyl-4-phosphonophenylglycine (MPPG; 250 microM). 3. The group I and II mGlu agonist, 1S,3R-aminocyclopentanedicarboxylic acid (ACPD; 3-30 microM) also depressed the e.p.s.p. in a concentration-dependent manner. The effect of ACPD (10 microM) was reversed by (+)-alpha-methyl-4-carboxyphenylglycine (MCPG; 1 mM; 4 cells). This effect of ACPD was also partially antagonized (by 50.3+/-15.7%, 4 cells) by MPPG (250 microM). 4. The selective agonist at group I mGlu receptors, dihydroxyphenylglycine (DHPG; 100 microM), decreased e.p.s.p. amplitude by 27.1+/-8.2% (7 cells), as did the group II mGlu receptor-selective agonist (1S,1R,2'R,3'R)-2-(2,3-dicarboxycyclopropyl)glycine (DCG-IV; 1 microM) by 26.7+/-4.3% (5 cells). 5. DHPG (10-100 microM) caused a depolarization of the recorded cell, as did ACPD (3-30 microM), whereas no such postsynaptic effect of either L-AP4 or DCG-IV was observed. 6. These results provide evidence for the presence of presynaptic inhibitory metabotropic glutamate autoreceptors from the mGlu receptor groups II and III on descending glutamatergic inputs to midbrain dopamine neurones. Group I mGlu receptors mediate a postsynaptic depolarization, and can also depress glutamatergic transmission, but may not necessarily be localized presynaptically. These sites represent novel drug targets for treatment of schizophrenia and movement disorders of basal ganglia origin.
Asunto(s)
Dopamina/metabolismo , Potenciales Postsinápticos Excitadores , Glutamatos/metabolismo , Mesencéfalo/fisiología , Neuronas/fisiología , Receptores de Glutamato Metabotrópico/fisiología , Alanina/análogos & derivados , Alanina/farmacología , Animales , Benzoatos/farmacología , Cicloleucina/análogos & derivados , Cicloleucina/farmacología , Estimulación Eléctrica , Glicina/análogos & derivados , Glicina/farmacología , Técnicas In Vitro , Masculino , Mesencéfalo/efectos de los fármacos , Mesencéfalo/metabolismo , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Ratas , Ratas Wistar , Receptores de Glutamato Metabotrópico/efectos de los fármacos , Receptores de Glutamato Metabotrópico/metabolismoRESUMEN
1. Intracellular recordings were made from presumed dopamine-containing neurones in the ventral tegmental area (VTA) in rat brain slices. 2. Nicotine (10-100 microM) and acetylcholine (ACh) depolarized the neurones. The depolarization caused by ACh was typically biphasic; both components were increased by neostigmine (0.1-10 microM), but only the slower component was blocked by scopolamine (1-10 microM). 3. The nicotinic action of ACh, studied in the presence of neostigmine and scopolamine, persisted in the presence of tetrodotoxin (1 microM) and cobalt (2-5 mM). 4. ACh or carbachol (30 microM) caused inward currents in neurones voltage-clamped near the resting potential. These currents reversed polarity at around -4 mV, were blocked by hexamethonium (1-100 microM) in a voltage-dependent manner, and showed desensitization with prolonged or repeated agonist applications. 5. Depolarizations caused by ACh and carbachol were reduced in slices pretreated with kappa-bungarotoxin, but were not changed by alpha-bungarotoxin. 6. These responses to ACh and nicotine resemble those previously described on autonomic ganglion cells. The direct action on VTA neurones may contribute to the positive reinforcement associated with nicotine consumption.
Asunto(s)
Neuronas/fisiología , Receptores Nicotínicos/fisiología , Tegmento Mesencefálico/fisiología , Acetilcolina/farmacología , Animales , Bungarotoxinas/farmacología , Carbacol/farmacología , Electrofisiología , Compuestos de Hexametonio/farmacología , Técnicas In Vitro , Masculino , Potenciales de la Membrana/efectos de los fármacos , Neuronas/efectos de los fármacos , Nicotina/farmacología , Ratas , Tegmento Mesencefálico/citologíaRESUMEN
1. Intracellular recordings were made from neurons in striatum (caudate-putamen) and substantia nigra pars compacta in rat brain slices. Three GABAB agonists, baclofen, 3-aminopropylphosphinic acid (3-APPA) and 3-aminopropyl(methyl)phosphinic acid (SK&F 97541), depressed excitatory postsynaptic potentials (e.p.s.ps) mediated by glutamate in the striatum, and hyperpolarized neurones in the substantia nigra. The ability of 3-aminopropyl(diethyoxymethyl)phosphinic acid (CGP 35348), 3-aminopropyl (hexyl)phosphinic acid (3-APHPA) and phaclofen to antagonize these responses was assessed. 2. Striatal e.p.s.ps, studied in the presence of bicuculline (30 microns), were reduced in amplitude by 92% with 6,7-dinitroquinoxaline-2,3-dione (DNQX; 30 microns). These e.p.s.ps were depressed by up to 95% by SK&F 97541 and baclofen with EC50s of 0.092 microns and 1.25 microns respectively. The maximal effect of 3-APPA was 67% with an EC50 of 0.83 microns. Agonist concentration-effect data fitted a single-site logistic model. GABAB agonists were without effect on striatal neurone membrane potential, input resistance or depolarizations induced by applied glutamate. 3. The depression of striatal e.p.s.ps by SK&F 97541 was reversibly antagonized by CGP 35348, 3-APHPA and phaclofen with estimated equilibrium dissociation constants (KB) of 11.2 +/- 1.7 microns (n = 4), 13.3 +/- 0.4 microM (n = 3) and 405 +/- 43 microM (n = 3) respectively. CGP 35348 and 3-APHPA appeared to act competitively (Schild plot slopes of 0.99 and 1.01 respectively). 4. Nigral neurones were hyperpolarized by up to 25 mV by SK&F 97541 and baclofen with EC50s of 0.15 microns and 3.6 microns respectively. The maximum hyperpolarization by 3-APPA was only 84% that of the other agonists, with an EC50 of 9.0 microM. Agonist concentration-effect data fitted a single-site logistic model. 5. The SK&F 97541-induced hyperpolarization was reversibly antagonized by CGP 35348, 3-APHPA and phaclofen with estimated KBS of 17.6 + 4.4 (n = 3), 14.0 + 1.5 (n = 4), and >400 microM (n = 1) respectively. CGP 35348 appeared competitive (Schild plot slope of 0.99). Antagonists were also tested with baclofen as agonist, yielding similar KB estimates as for SK&F 97541. 6. It is concluded that at both the presynaptic and postsynaptic sites examined, SK&F 97541 was about 10 fold more potent than baclofen or 3-APPA. The antagonists CGP 35348 and 3-APHPA (KB 1O-20 microM) were about 20 fold more potent than phaclofen. The similarities in relative agonist potency and estimated antagonist affinity between these two functionally distinct GABAB receptors renders them pharmacologically indistinguishable at present.
Asunto(s)
Encéfalo/efectos de los fármacos , Neuronas/efectos de los fármacos , Receptores de GABA-A/efectos de los fármacos , Animales , Baclofeno/análogos & derivados , Baclofeno/farmacología , Bicuculina/farmacología , Encéfalo/citología , Cuerpo Estriado/efectos de los fármacos , Cuerpo Estriado/fisiología , Electrofisiología , Antagonistas de Receptores de GABA-A , Técnicas In Vitro , Potenciales de la Membrana/efectos de los fármacos , Compuestos Organofosforados/farmacología , Ratas , Sustancia Negra/efectos de los fármacos , Sustancia Negra/fisiología , Sinapsis/efectos de los fármacosRESUMEN
1. Intracellular recordings were made from neurones in vitro in the rat substantia nigra zona compacta and ventral tegmental area; the neurones had the characteristic properties of dopamine-containing cells, and fired action potentials spontaneously. 2. Cocaine (1-10 microM) inhibited spontaneous firing, hyperpolarized the membrane and (in neurones voltage-clamped at -60 mV) caused an outward membrane current; the minimally effective concentration was 1 microM. These effects were blocked by sulpiride (30 nM-1 microM). 3. Dopamine (3-100 microM) also inhibited firing, hyperpolarized and caused an outward current. These effects of dopamine were potentiated about five fold by cocaine (10 microM). 4. It is concluded that cocaine (1-10 microM) inhibits the firing and hyperpolarizes substantia nigra zona compacta neurones in vitro by blocking the uptake of dopamine which the cells continuously release.
Asunto(s)
Cocaína/farmacología , Dopamina/fisiología , Neuronas/metabolismo , Potenciales de Acción/efectos de los fármacos , Animales , Dopamina/farmacología , Técnicas In Vitro , Microelectrodos , Neuronas/efectos de los fármacos , Ratas , Sustancia Negra/efectos de los fármacos , Sulpirida/farmacología , Tetrodotoxina/farmacologíaRESUMEN
Extracellular single unit recordings were made from substantia nigra pars reticulata (SNr) neurones in slices of rat brain. Cells fired spontaneous action potentials at 11.4 +/- 0.8 Hz. The GABAA receptor agonist isoguvacine (1-10 microM) reduced firing rate in a concentration-dependent manner [50% of maximal inhibition (IC50) with 3.2 microM], as did the GABAB agonist baclofen (0.3-10 microM; IC50 1.4 microM). The GABAA antagonist bicuculline (30 microM) not only blocked the action of isoguvacine, but also increased the basal firing rate to 187.5 +/- 12.6% of control. The GABAB antagonist CGP 55845A (0.1 microM), while blocking the inhibitory action of baclofen, was without effect on spontaneous firing rate, as was strychnine (10 microM), the antagonist of glycine and taurine, and also Met-enkephalin (10 microM). Tiagabine (50 microM), the blocker of GABA uptake, caused an inhibition of firing which could be reversed with bicuculline (30 microM) but not CGP 55845A (1 microM). We conclude that the firing rate of SNr neurones is under tonic inhibition by GABA in vitro, which can be relieved by antagonists of GABAA, but not GABAB receptors, and enhanced by blockade of GABA reuptake. The source of this GABA tone is likely to be from recurrent axon collaterals of SNr neurones themselves.
Asunto(s)
Inhibición Neural/fisiología , Neuronas/fisiología , Receptores de GABA/fisiología , Sustancia Negra/fisiología , Animales , Encefalina Metionina/farmacología , Agonistas del GABA/farmacología , Antagonistas del GABA/farmacología , Técnicas In Vitro , Masculino , Neuronas/efectos de los fármacos , Ácidos Nipecóticos/farmacología , Ratas , Ratas Sprague-Dawley , Receptores de GABA/clasificación , Sustancia Negra/citología , Sustancia Negra/efectos de los fármacos , TiagabinaRESUMEN
gamma-Aminobutyric acid (GABA)-mediated synaptic potentials in rat dorsal striatum in vitro were reduced in amplitude by the GABAB receptor agonists baclofen and 3-aminopropyl-(methyl)-phosphinic acid (SK&F 97541), without any detectable postsynaptic effect. Synaptic potentials in 40% of neurones were distinctly multiphasic, the components of which exhibited a differential sensitivity to GABAB agonists. One population of GABA-releasing neurones within the striatum had presynaptic GABAB autoreceptors, whereas others were not directly affected by GABAB agonists.
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Núcleo Caudado/fisiología , Putamen/fisiología , Receptores de GABA-A/fisiología , Membranas Sinápticas/fisiología , Ácido gamma-Aminobutírico/fisiología , Animales , Baclofeno/farmacología , Técnicas In Vitro , Potenciales de la Membrana/fisiología , Compuestos Organofosforados/farmacología , Ratas , Receptores de GABA-A/efectos de los fármacosRESUMEN
The subthalamic nucleus (SThN) provides a glutamate mediated excitatory drive to several other component nuclei of the basal ganglia, thereby significantly influencing locomotion and control of voluntary movement. We have characterised functionally the metabotropic glutamate (mGlu) receptors in the SThN using extracellular single unit recording from rat midbrain slices. SThN neurones fired action potentials spontaneously at a rate of 10 Hz which was increased by the group I/II mGlu receptor agonist (1S,3R)-1-aminocyclopentane-1,3-dicarboxylate (1S,3 R-ACPD; 1-30 microM) and the group I selective agonist (S, R)-dihydroxyphenylglycine (DHPG; 1-30 microM). However, both the group II selective agonist (1S,1'R,2'R,3'R)-2-(2,3-dicarboxycyclopropyl)glycine (DCG-IV; 1 microM) and the group III selective agonist (S)-2-amino-4-phosphonobutanoic acid (L-AP4; 10 microM) were without effect, indicating that the excitation was mediated by a group I mGlu receptor. The excitation caused by DHPG (3 microM) was reversed by co-application of the mGlu receptor antagonist (+)-alpha-methyl-4-carboxyphenylglycine (MCPG; 500 microM). Thus a group I mGlu receptor mediates excitation of SThN neurones, and suggests a use for group I mGlu receptor ligands for treatment of both hypo- and hyperkinetic disorders of basal ganglia origin, such as Parkinson's disease and Huntington's disease.
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Neuronas/química , Receptores de Glutamato Metabotrópico/fisiología , Núcleos Talámicos/citología , Nucleótidos de Adenina/farmacología , Marcadores de Afinidad/farmacología , Animales , Benzoatos/farmacología , Cicloleucina/análogos & derivados , Cicloleucina/farmacología , Ciclopropanos/farmacología , Electrofisiología , Antagonistas de Aminoácidos Excitadores/farmacología , Glicina/análogos & derivados , Glicina/farmacología , Masculino , Neuronas/efectos de los fármacos , Neuronas/fisiología , Fármacos Neuroprotectores/farmacología , Piridoxal/análogos & derivados , Piridoxal/farmacología , Ratas , Ratas Wistar , Receptores de Glutamato Metabotrópico/agonistas , Receptores de Glutamato Metabotrópico/antagonistas & inhibidores , Resorcinoles/farmacología , Núcleos Talámicos/fisiologíaRESUMEN
Intracellular recordings were made from substantia nigra dopamine neurones in a rat brain slice preparation. Spontaneous firing in these cells was reversibly inhibited by taurine applied by superfusion (300 microM-20 mM) or by focal pressure ejection. Neurones recorded with electrodes filled with KCl were depolarised at resting potential by taurine; the taurine depolarisation reversed polarity at -36.6 +/- 1.0 mV (7 cells). When electrodes filled with K-acetate or K-methyl sulphate were used, taurine caused a hyperpolarisation which reversed at -74.2 +/- 3.8 mV (9 cells). These effects of taurine were accompanied by a fall in input resistance or, in voltage clamp, an increase in conductance. Taurine thus appeared to increase membrane chloride conductance. The effect of taurine persisted in tetrodotoxin, 0-Ca2+/10 mM Mg2+, and bicuculline, but was blocked by strychnine (10 microM). Maximal responses to either taurine or glycine occluded responses to the other amino acid. Taurine therefore acts directly on dopamine neurones in the substantia nigra to increase the same membrane Cl- conductance as that mediating the action of glycine. Taurine may also act at the same recognition site as glycine in these cells.
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Cloruros/fisiología , Glicina/farmacología , Neuronas/metabolismo , Sustancia Negra/metabolismo , Taurina/farmacología , Animales , Conductividad Eléctrica , Electrofisiología , Técnicas In Vitro , Ratas , Sustancia Negra/citologíaAsunto(s)
Canales Iónicos/fisiología , Neuronas/fisiología , Receptores de Neurotransmisores/fisiología , Sustancia Negra/fisiología , Área Tegmental Ventral/fisiología , Acetilcolina/farmacología , Animales , Monoaminas Biogénicas/farmacología , Monoaminas Biogénicas/fisiología , Dopamina/fisiología , Mesencéfalo/fisiología , Neuronas/efectos de los fármacos , Ratas , Ácido gamma-Aminobutírico/farmacologíaAsunto(s)
Cuerpo Estriado/fisiología , Receptores de GABA-A/fisiología , Sinapsis/fisiología , Potenciales de Acción/efectos de los fármacos , Animales , Baclofeno/farmacología , Cuerpo Estriado/efectos de los fármacos , Antagonistas de Receptores de GABA-A , Neuronas/efectos de los fármacos , Neuronas/fisiología , Compuestos Organofosforados/farmacología , RatasAsunto(s)
Dinorfinas/farmacología , Fragmentos de Péptidos/farmacología , Pirrolidinas/farmacología , Receptores Opioides/efectos de los fármacos , Conducto Deferente/efectos de los fármacos , 3,4-Dicloro-N-metil-N-(2-(1-pirrolidinil)-ciclohexil)-bencenacetamida, (trans)-Isómero , Animales , Electrofisiología , Ganglios/efectos de los fármacos , Técnicas In Vitro , Masculino , Conejos , Receptores Opioides kappa , Transmisión Sináptica/efectos de los fármacos , Conducto Deferente/fisiologíaRESUMEN
In accordance with its central role in basal ganglia circuitry, changes in the rate of action potential firing and pattern of activity in the globus pallidus (GP)-subthalamic nucleus (STN) network are apparent in movement disorders. In this study we have developed a mouse brain slice preparation that maintains the functional connectivity between the GP and STN in order to assess its role in shaping and modulating bursting activity promoted by pharmacological manipulations. Fibre-tract tracing studies indicated that a parasagittal slice cut 20 deg to the midline best preserved connectivity between the GP and the STN. IPSCs and EPSCs elicited by electrical stimulation confirmed connectivity from GP to STN in 44/59 slices and from STN to GP in 22/33 slices, respectively. In control slices, 74/76 (97%) of STN cells fired tonically at a rate of 10.3 +/- 1.3 Hz. This rate and pattern of single spiking activity was unaffected by bath application of the GABA(A) antagonist picrotoxin (50 microM, n = 9) or the glutamate receptor antagonist (6-cyano-7-nitroquinoxaline-2, 3-dione (CNQX) 10 microM, n = 8). Bursting activity in STN neurones could be induced pharmacologically by application of NMDA alone (20 microm, 3/18 cells, 17%) but was more robust if NMDA was applied in conjunction with apamin (20-100 nM, 34/77 cells, 44%). Once again, neither picrotoxin (50 microM, n = 5) nor CNQX (10 microM, n = 5) had any effect on the frequency or pattern of the STN neurone activity while paired STN and GP recordings of tonic and bursting activity show no evidence of coherent activity. Thus, in a mouse brain slice preparation where functional GP-STN connectivity is preserved, no regenerative synaptically mediated activity indicative of a dynamic network is evident, either in the resting state or when neuronal bursting in both the GP and STN is generated by application of NMDA/apamin. This difference from the brain in Parkinson's disease may be attributed either to insufficient preservation of cortico-striato-pallidal or cortico-subthalamic circuitry, and/or an essential requirement for adaptive changes resulting from dopamine depletion for the expression of network activity within this tissue complex.