RESUMEN
Although dopamine (DA) effects in the prefrontal cortex (PFC) have been studied extensively, the function of steady-state ambient levels of DA in the regulation of afferent excitatory transmission in PFC pyramidal neurons remains relatively unexplored. Using intracellular sharp-electrode and whole-cell recordings combined with intracellular labeling in brain slices, we found that D1/D5 receptor blockade did not alter synaptic responses in the PFC, but D1/D5 receptor activation consistently enhanced recurrent synaptic excitation in the majority of pyramidal neurons tested. In contrast, D4 receptor blockade resulted in an evoked complex multiple spike discharge pattern that contained both early and late (presumably multisynaptic) components of the evoked response that is contingent upon the preservation of axon collaterals of the neuron under study. Moreover, GABAergic interneurons were found to play a role in both responses; blockade of GABA(a)-mediated inhibition caused bath application of DA to convert monosynaptic excitatory postsynaptic potentials (EPSPs) to complex spike bursts riding on the late component of the EPSP. On the other hand, during the blockade of GABA(a)-mediated conductances, administration of a D4 receptor antagonist failed to facilitate evoked multiple spike discharge. Morphological analysis of axon collaterals of labeled neurons revealed that neurons in which the D4 receptor blockade induced the putative polysynaptic response had axon collaterals that were largely preserved. These data suggest that DA exerts a bidirectional modulation of PFC pyramidal neurons in brain slices provided that local network connections with interneurons are preserved, with D4 receptors under tonic stimulation by ambient low levels of DA, whereas D1/D5 receptors activated upon phasic DA input.
Asunto(s)
Corteza Prefrontal/citología , Células Piramidales/fisiología , Receptores de Dopamina D1/fisiología , Receptores de Dopamina D4/fisiología , Sinapsis/fisiología , Animales , Bicuculina/farmacología , Dopamina/metabolismo , Agonistas de Dopamina/farmacología , Antagonistas de Dopamina/farmacología , Relación Dosis-Respuesta en la Radiación , Interacciones Farmacológicas , Impedancia Eléctrica , Estimulación Eléctrica/métodos , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Potenciales Postsinápticos Excitadores/fisiología , Potenciales Postsinápticos Excitadores/efectos de la radiación , Antagonistas del GABA/farmacología , Técnicas In Vitro , Lisina/análogos & derivados , Masculino , Potenciales de la Membrana/efectos de los fármacos , Potenciales de la Membrana/fisiología , Potenciales de la Membrana/efectos de la radiación , Técnicas de Placa-Clamp/métodos , Picrotoxina/farmacología , Células Piramidales/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Receptores de Dopamina D1/agonistas , Receptores de Dopamina D1/antagonistas & inhibidores , Receptores de Dopamina D4/agonistas , Receptores de Dopamina D4/antagonistas & inhibidores , Sinapsis/efectos de los fármacosRESUMEN
We have recently shown that chronic amphetamine exposure selectively up-regulates parvalbumin (PV) calcium-binding proteins in the anterior cingulate cortex (ACC). In this study, we evaluated the effects of chronic nicotine (NIC) exposure on PV, calbindin D28k (CB) and calretinin (CR) calcium-binding protein immunostaining in ACC GABAergic interneurons. Chronic NIC exposure for 3 weeks in adolescent rats, either via drinking water (the oral group) or by twice daily subcutaneous injections (the injection group), resulted in the expression of high levels of CR proteins in the ACC but not in the parietal cortex. Larger increases in the density of CR-immunoreactive (ir) neurons were noted in the NIC-injected rats at 0-day withdrawal (45% increase) compared with the oral group (26% increase). The larger increases in CR-ir neuron density in the NIC-injected rats were also reflected by prominent CR-ir processes across cortical layers. The density of PV-ir neurons was also increased (37%) at 0-day withdrawal but only in the oral NIC group and no changes in CB-ir neuron density were observed in either NIC group. Combined dual-immunofluorescence and confocal microscopy revealed that somatodendritic alpha4 nicotinic acetylcholine receptors colocalized with cortical neurons stained positively for CR, PV or CB. These results suggest that CR- and/or PV-ir-containing GABA interneurons may be involved in channeling the effects of NIC in the ACC, which is closely associated with the ventral basal ganglia circuit that is linked to brain reward function.
Asunto(s)
Proteínas de Unión al Calcio/biosíntesis , Giro del Cíngulo/metabolismo , Nicotina/farmacología , Agonistas Nicotínicos/farmacología , Administración Oral , Envejecimiento/fisiología , Animales , Técnica del Anticuerpo Fluorescente , Giro del Cíngulo/citología , Giro del Cíngulo/efectos de los fármacos , Técnicas para Inmunoenzimas , Inmunohistoquímica , Inyecciones Subcutáneas , Interneuronas/fisiología , Masculino , Microscopía Confocal , Nicotina/administración & dosificación , Agonistas Nicotínicos/administración & dosificación , Lóbulo Parietal/efectos de los fármacos , Lóbulo Parietal/fisiología , Ratas , Ratas Sprague-Dawley , Ácido gamma-Aminobutírico/fisiologíaRESUMEN
A distinct increase in cell firing activity is reported in prefrontal cortex during working memory tasks. The afferents that modulate this activity are not yet identified. Using in vivo intracellular recording and labelling of prefrontal cortical pyramidal neurons in anaesthetized rats, we systematically evaluated the influences of afferent projections arising from the ventral tegmental area (VTA) and mediodorsal thalamus (MD) by phasic electrical stimulation with a range of stimulus frequencies. Both VTA- and MD-responsive pyramidal neurons exhibited extensive intracortical axon arborization. Neither single shocks to the VTA at 0.2 Hz, nor low frequency trains of stimuli at 1-4 Hz (< 5 Hz) interrupted the periodicity of membrane bistability in bistable pyramidal neurons. However, high-frequency VTA-train stimulation (10-50 Hz) interrupted the bistability, and produced sustained membrane depolarizations accompanied by intense tonic firing in a frequency-dependent manner. Electrical stimulation of MD (10-50 Hz) did not produce sustained activity in the same PFC neurons. Thus, the sustained activity induced by high-frequency VTA trains is input selective. This effect of VTA-train stimulation was attenuated by systemic injection of the D1 receptor antagonist, SCH 23390, and blocked by acute dopamine (DA) depletion produced via alpha-methyl-para-tyrosine pre-treatment, suggesting that sustained cortical activity is mediated by DA. Chemical stimulation of VTA via intra-VTA infusion of NMDA induced sustained activity similar to VTA-train stimulation. Thus, while both VTA- and MD-responsive pyramidal neurons exhibited extensive intracortical axon arborization, VTA synapses (as opposed to MD synapses) may be critically positioned in the dendritic arborizations of anterior cingulate cortical pyramidal neurons, which may allow their modulation of sustained activity in prefrontal bistable neurons.
Asunto(s)
Dopamina/metabolismo , Giro del Cíngulo/fisiología , Núcleo Talámico Mediodorsal/fisiología , Vías Nerviosas/fisiología , Células Piramidales/fisiología , Área Tegmental Ventral/fisiología , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/fisiología , Animales , Dendritas/efectos de los fármacos , Dendritas/fisiología , Antagonistas de Dopamina/farmacología , Estimulación Eléctrica , Agonistas de Aminoácidos Excitadores/farmacología , Giro del Cíngulo/efectos de los fármacos , Lisina/análogos & derivados , Masculino , Vías Nerviosas/efectos de los fármacos , Terminales Presinápticos/efectos de los fármacos , Terminales Presinápticos/fisiología , Células Piramidales/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Receptores Dopaminérgicos/efectos de los fármacos , Receptores Dopaminérgicos/metabolismo , Transmisión Sináptica/efectos de los fármacos , Transmisión Sináptica/fisiología , Área Tegmental Ventral/efectos de los fármacos , alfa-Metiltirosina/farmacologíaRESUMEN
We previously reported synchronization of pyramidal neurons within prefrontal cortex of rats repeatedly exposed to amphetamine (AMPH). To test the hypothesis that cortical synchronization may be related to changes in local GABA signaling, we used antibodies specific for parvalbumin (PV), calbindin D28k (CB) and calretinin (CR) as selective labels for three distinct GABA interneuron classes in the anterior cingulate cortex (ACC) of similarly treated rats. We observed a selective increase in the density of PV-immunoreactive (ir), but not CB-ir or CR-ir, neurons in the ACC of AMPH-treated rats at both 1 day and 7 day withdrawal. Increased density of PV-ir GABA interneurons in the ACC at 1 day withdrawal was reproduced in rats repeatedly injected with apomorphine or with SKF-38393. Thus, the critical role of DA receptors during AMPH exposure is evident. However, DA receptor activation did not appear to account for the PV up-regulation in AMPH-treated rats at 7 day withdrawal. Significantly higher numbers of pericellular basket-like puncta immunoreactive for corticotropin-releasing factor (CRF) were observed in the ACC of AMPH rats at 7 day withdrawal. Combined dual immunofluorescence and confocal microscopy further revealed that CRF-ir puncta made possible pericellular contacts on PV-ir (not CB-, CR- or glutamate-ir) cell bodies. A potential cellular mechanism seems to emerge that CRF-ir terminals, that may be underdetected under normal conditions due to low activity levels, may be functionally activated during psychostimulant withdrawal, thereby altering local GABAergic signaling.
Asunto(s)
Anfetamina/efectos adversos , Proteínas de Unión al Calcio/metabolismo , Hormona Liberadora de Corticotropina/metabolismo , Lóbulo Frontal/metabolismo , Lóbulo Frontal/patología , Interneuronas/metabolismo , Síndrome de Abstinencia a Sustancias/metabolismo , Ácido gamma-Aminobutírico/metabolismo , Anfetamina/administración & dosificación , Trastornos Relacionados con Anfetaminas/metabolismo , Trastornos Relacionados con Anfetaminas/patología , Animales , Calbindina 1 , Calbindina 2 , Calbindinas , Recuento de Células , Relación Dosis-Respuesta a Droga , Giro del Cíngulo/metabolismo , Giro del Cíngulo/patología , Interneuronas/patología , Masculino , Parvalbúminas/metabolismo , Ratas , Ratas Sprague-Dawley , Proteína G de Unión al Calcio S100/metabolismo , Transducción de Señal , Síndrome de Abstinencia a Sustancias/patologíaRESUMEN
The phosphoprotein DARPP-32 (dopamine and cAMP-regulated phosphoprotein 32 kDa) plays a central role in mediating the actions of a variety of neurotransmitters in medium spiny neurons of the striatum (Greengard, 1990; Fienberg et al., 1998). This study examines D1 and D2 dopamine (DA) agonist effects on the membrane properties of identified striatal neurons recorded in slices obtained from wild-type and DARPP-32-knockout mice. In wild-type spiny cells, DA D1 receptor activation decreased cell excitability, causing a 58.8 +/- 13.5% increase in rheobase current required to evoke spike discharge. In contrast, D1 agonist administration did not alter cell excitability when applied to spiny cells in slices prepared from the DARPP-32 knockout mice. D2 agonist administration decreased cell excitability in both wild-type and knockout mice. The response produced by combined D1 and D2 agonist stimulation was dependent on the sequence of agonist administration. Thus, the D1 agonist-induced decrease in excitability was reversed to a facilitation of spiking upon subsequent D2 agonist administration. In contrast, D2 agonist applied simultaneously with the D1 agonist only produced a reduction in excitability. This type of D1-dependent modulation was not present in slices from the DARPP-32 knockout mice.