RESUMEN
Mastication can be triggered by repetitive stimulation of the cortex or of sensory inputs, but is patterned by a brain stem central pattern generator (CPG). This CPG may include the dorsal part of the principal trigeminal sensory nucleus (NVsnpr), where neurons burst repetitively when the extracellular concentration of Ca(2+) ([Ca(2+)](e)) drops. We examined the effects of repetitive stimulation of sensory afferents of the trigeminal tract on activity of NVsnpr neurons recorded extracellularly in vitro under physiologic [Ca(2+)](e) (1.6 mM). Spontaneously active cells had either a tonic (n = 145) or a bursting (n = 46) firing pattern. Afferent stimulation altered burst duration and/or burst frequency in bursting cells and firing frequency in most tonic cells. In 28% of the latter, the firing pattern switched to rhythmic bursting. This effect could be mimicked by local application of N-methyl-d-aspartate and blocked by APV but not DNQX. Detailed analysis showed that rhythm indices (RIs) of 35 tonic neurons that were negative (nonrhythmic) before stimulation became significantly rhythmic (RI > or = 0.01) after stimulation. Mean and median bursting frequency of these units were 8.32 +/- 0.72 (SE) Hz and 6.25 Hz (range, 2.5-17.5 Hz). In seven instances, two units were recorded simultaneously, and cross-correlation analysis showed that firing of six pairs was rhythmic and synchronized after stimulation. Optimal stimulation parameters for eliciting rhythmic bursting consisted in 500-ms trains of pulses delivered at 40-60 Hz. Together, our results show that repetitive stimulation of sensory afferents in vitro can elicit masticatory-like rhythmic bursting in NVsnpr neurons at physiological [Ca(2+)](e).
Asunto(s)
Potenciales de Acción/fisiología , Relojes Biológicos/fisiología , Señalización del Calcio/fisiología , Calcio/administración & dosificación , Calcio/metabolismo , Células Receptoras Sensoriales/fisiología , Núcleos del Trigémino/fisiología , Potenciales de Acción/efectos de los fármacos , Animales , Relojes Biológicos/efectos de los fármacos , Señalización del Calcio/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Células Receptoras Sensoriales/efectos de los fármacos , Núcleos del Trigémino/efectos de los fármacosRESUMEN
Brainstem networks generating the respiratory rhythm in lampreys are still not fully characterized. In this study, we described the patterns of respiratory activities and we identified the general location of underlying neural networks. In a semi-intact preparation including the brain and gills, rhythmic discharges were recorded bilaterally with surface electrodes placed over the vagal motoneurons. The main respiratory output driving rhythmic gill movements consisted of short bursts (40.9+/-15.6 ms) of discharge occurring at a frequency of 1.0+/-0.3 Hz. This fast pattern was interrupted by long bursts (506.3+/-174.6 ms) recurring with an average period of 37.4+/-24.9 s. After isolating the brainstem by cutting all cranial nerves, the frequency of the short respiratory bursts did not change significantly, but the slow pattern was less frequent. Local injections of a glutamate agonist (AMPA) and antagonists (6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) or D,L-amino-5-phosphonopentanoic acid (AP5)) were made over different brainstem regions to influence respiratory output. The results were similar in the semi-intact and isolated-brainstem preparations. Unilateral injection of AP5 or CNQX over a rostral rhombencephalic region, lateral to the rostral pole of the trigeminal motor nucleus, decreased the frequency of the fast respiratory rhythm bilaterally or stopped it altogether. Injection of AMPA at the same site increased the rate of the fast respiratory rhythm and decreased the frequency of the slow pattern. The activity recorded in this area was synchronous with that recorded over the vagal motoneurons. After a complete transverse lesion of the brainstem caudal to the trigeminal motor nucleus, the fast rhythm was confined to the rostral area, while only the slow activity persisted in the vagal motoneurons. Our results support the hypothesis that normal breathing depends on the activity of neurons located in the rostral rhombencephalon in lampreys, whereas the caudal rhombencephalon generates the slow pattern.
Asunto(s)
Red Nerviosa/fisiología , Vías Nerviosas/fisiología , Petromyzon/fisiología , Centro Respiratorio/fisiología , Fenómenos Fisiológicos Respiratorios/efectos de los fármacos , Rombencéfalo/fisiología , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/fisiología , Animales , Relojes Biológicos/efectos de los fármacos , Relojes Biológicos/fisiología , Región Branquial/inervación , Región Branquial/fisiología , Agonistas de Aminoácidos Excitadores/farmacología , Antagonistas de Aminoácidos Excitadores/farmacología , Femenino , Branquias/inervación , Branquias/fisiología , Ácido Glutámico/metabolismo , Masculino , Bulbo Raquídeo/anatomía & histología , Bulbo Raquídeo/efectos de los fármacos , Bulbo Raquídeo/fisiología , Neuronas Motoras/efectos de los fármacos , Neuronas Motoras/fisiología , Músculo Esquelético/inervación , Músculo Esquelético/fisiología , Red Nerviosa/anatomía & histología , Red Nerviosa/efectos de los fármacos , Vías Nerviosas/anatomía & histología , Vías Nerviosas/efectos de los fármacos , Periodicidad , Petromyzon/anatomía & histología , Puente/anatomía & histología , Puente/efectos de los fármacos , Puente/fisiología , Centro Respiratorio/anatomía & histología , Centro Respiratorio/efectos de los fármacos , Rombencéfalo/anatomía & histología , Rombencéfalo/efectos de los fármacos , Transmisión Sináptica/efectos de los fármacos , Transmisión Sináptica/fisiología , Factores de Tiempo , Nervio Vago/efectos de los fármacos , Nervio Vago/fisiologíaRESUMEN
The essential elements controlling trigeminal motoneurons during feeding lie between the trigeminal and facial motor nuclei. These include populations of neurons in the medial reticular formation and pre-motoneurons in the lateral brainstem that reorganize to generate various patterns. Orofacial sensory feedback, antidromic firing in spindle afferents and intrinsic properties of motoneurons also contribute to the final masticatory motor output.
Asunto(s)
Tronco Encefálico/fisiología , Conducta Alimentaria/fisiología , Animales , Masticación/fisiología , Modelos Neurológicos , Neuronas Motoras/fisiología , Vías Nerviosas/fisiología , Neuronas Aferentes/fisiología , Neurotransmisores/fisiología , Periodicidad , Nervio Trigémino/citología , Nervio Trigémino/fisiologíaRESUMEN
Three series of experiments were carried out to characterize interneurons located within the trigeminal motor nucleus of young rats aged 5-24 days. Cholera toxin injections were made bilaterally into the masseter and, sometimes, digastric muscles to label motoneurons. In the first set of experiments, thick slices were taken from the pontine brainstem and cholera toxin-positive and cholera toxin-negative neurons located inside the trigeminal motor nucleus were filled with biocytin through whole-cell recording patch electrodes. Positively identified motoneurons (cholera toxin+) of various shapes and sizes always had a thick, unbranched axon that entered the motor root following a tight zigzag course. Many cholera toxin-negative neurons were also classified as motoneurons after biocytin filling based on this particularity of their axon. These are probably either fusimotor motoneurons or motoneurons supplying other jaw muscles. The cholera toxin-negative neurons classified as interneurons differed markedly from motoneurons in that they had thin, usually branched axons that supplied the ipsilateral reticular region surrounding the trigeminal motor nucleus (peritrigeminal area), the main trigeminal sensory nucleus, the trigeminal mesencephalic nucleus, the medial reticular formation of both sides, and the contralateral medial peritrigeminal area. Most often, their dendrites were arranged in bipolar arbors that extended beyond the borders of the trigeminal motor nucleus into the peritrigeminal area. Immunohistochemistry against glutamate, GABA and glycine was used to further document the nature and distribution of putative interneurons. Immunoreactive neurons were uniformly distributed throughout the rostro-caudal extent of the trigeminal motor nucleus. Their concentration seemed greater toward the edges of the nucleus and they were scarce in the digastric motoneuron pool. Glutamate- outnumbered GABA- and glycine-immunoreactive neurons. There was no clear segregation between the three populations. In the final experiment, 1,1'-dioctadecyl-3,3,3',3'-tetra-methylindocarbocyanine perchlorate crystals were inserted into one trigeminal motor nucleus in thick slices and allowed to diffuse for several weeks. This procedure marked commissural fibers and interneurons in the contralateral trigeminal motor nucleus. Together these results conclusively support the existence of interneurons in the trigeminal motor nucleus.
Asunto(s)
Interneuronas/citología , Músculo Masetero/inervación , Vías Nerviosas/citología , Núcleos del Trigémino/citología , Animales , Ácido Glutámico/metabolismo , Glicina/metabolismo , Inmunohistoquímica , Interneuronas/metabolismo , Neuronas Motoras/citología , Ratas , Ratas Sprague-Dawley , Núcleos del Trigémino/metabolismo , Ácido gamma-Aminobutírico/metabolismoRESUMEN
Dendritic defects occur in neurodegenerative diseases accompanied by axonopathy, yet the mechanisms that regulate these pathologic changes are poorly understood. Using Thy1-YFPH mice subjected to optic nerve axotomy, we demonstrate early retraction of retinal ganglion cell (RGC) dendrites and selective loss of mammalian target of rapamycin (mTOR) activity, which precede soma loss. Axonal injury triggered rapid upregulation of the stress-induced protein REDD2 (regulated in development and DNA damage response 2), a potent inhibitor of mTOR. Short interfering RNA-mediated REDD2 knockdown restored mTOR activity and rescued dendritic length, area and branch complexity in a rapamycin-dependent manner. Whole-cell recordings demonstrated that REDD2 depletion leading to mTOR activation in RGCs restored their light response properties. Lastly, we show that REDD2-dependent mTOR activity extended RGC survival following axonal damage. These results indicate that injury-induced stress leads to REDD2 upregulation, mTOR inhibition and dendrite pathology causing neuronal dysfunction and subsequent cell death.
Asunto(s)
Axones/metabolismo , Dendritas/fisiología , Traumatismos del Nervio Óptico/patología , Proteínas/metabolismo , Serina-Treonina Quinasas TOR/metabolismo , Proteínas Adaptadoras Transductoras de Señales , Animales , Apoptosis , Proteínas de Unión al ADN , Dendritas/efectos de los fármacos , Inmunosupresores/farmacología , Ratones , Ratones Transgénicos , Traumatismos del Nervio Óptico/metabolismo , Técnicas de Placa-Clamp , Proteínas/antagonistas & inhibidores , Proteínas/genética , Interferencia de ARN , ARN Interferente Pequeño/metabolismo , Células Ganglionares de la Retina/citología , Células Ganglionares de la Retina/metabolismo , Sirolimus/farmacología , Serina-Treonina Quinasas TOR/antagonistas & inhibidores , Antígenos Thy-1/genética , Factores de Transcripción , Regulación hacia ArribaRESUMEN
Recent studies indicate that long-term potentiation is accompanied by changes in the waveform of field excitatory postsynaptic potentials in the CA1 field of disinhibited hippocampal slices, suggesting that long-term potentiation alters the kinetics of the glutamate receptor channels that mediate excitatory synaptic responses. The present study examined the effects of stimulating and recording location within stratum radiatum on the magnitude of long-term potentiation and the associated waveform changes. Stimulation of stratum radiatum proximal to the cell body layer evoked field excitatory postsynaptic potentials in proximal stratum radiatum that had decay phases well-fit by single exponentials; long-term potentiation reduced the decay time constant of these potentials, as reported previously. Stimulation of distal stratum radiatum evoked field excitatory postsynaptic potentials in distal stratum radiatum that were contaminated by positive after-potentials; meaningful estimates of decay time constants for these responses could not be obtained. Long-term potentiation of distal responses tended to be smaller than that obtained proximally. Comparisons were also made between responses recorded distally and proximally for either distal or proximal stimulation. For both stimulation loci, the distally-recorded responses had positive after-potentials and the proximally-recorded responses did not. The decay time constants for proximally-recorded responses to distal stimulation decreased significantly after long-term potentiation. Proximal and distal stimulation both produced greater long-term potentiation recorded proximally than distally and the difference was significantly greater for proximal stimulation. When long-term potentiation was induced by stimulation of proximal and distal sites simultaneously, the difference between proximally- and distally-recorded long-term potentiation was significantly reduced. Paired-pulse facilitation was greater when recorded proximally but the variation in facilitation with stimulation and recording position was different from that obtained for long-term potentiation. Paired-pulse facilitation of response amplitude was slightly reduced after long-term potentiation, but the change did not depend on the stimulation-recording arrangement; facilitation of response slope was not reliably affected by long-term potentiation. These results indicate that the shape of the dendritic field excitatory postsynaptic potential is influenced by the relative position of recording electrodes in stratum radiatum; when the position is such that after-potentials are minimized, long-term potentiation produces a decrease in the decay time constant of the synaptic field potential.(ABSTRACT TRUNCATED AT 400 WORDS)
Asunto(s)
Dendritas/fisiología , Hipocampo/fisiología , Potenciación a Largo Plazo/fisiología , Animales , Electrofisiología , Hipocampo/citología , Potenciales de la Membrana/fisiología , Ratas , Ratas Sprague-Dawley , Factores de TiempoRESUMEN
The results of a previous experiment suggest that the cell bodies of many jaw closing muscle spindle afferents in the trigeminal mesencephalic nucleus of the rabbit are phasically inhibited during fictive mastication. The aim of this study was to investigate one possible neurotransmitter system that could be involved in this modulation, serotonin, by use of receptor autoradiography techniques and immunofluorescence combined with retrograde labelling of masseteric spindle afferents and motoneurons. A second objective was to compare the serotonin innervation of neurons in the trigeminal mesencephalic nucleus with that of masseteric motoneurons. Serotoninergic fibres were seen surrounding labelled masseteric spindle afferents, as well as unlabelled neurons, in the trigeminal mesencephalic nucleus. These fibres were close to the cell bodies and sometimes to the axon hillocks of the neurons. Although it has been reported that many neurons of the trigeminal nucleus are multipolar in some species, none of the labelled spindle afferent in this study had more than one process. Throughout the motor trigeminal nucleus, serotonin fibres were found in close proximity with cell bodies and with the proximal portions of axons and dendrites of labelled and unlabelled motoneurons. Serotonin fibres were also seen adjacent to cell bodies and processes of efferent neurons in cell group k. Autoradiography with several tritiated ligands was used to reveal the presence of receptors for serotonin as well as its uptake sites. Only serotonin2 receptors were found to be abundant in the trigeminal mesencephalic nucleus. The motor nucleus and cell group k contained serotonin2 and serotonin3 receptors, as well as serotonin uptake sites. Serotonin1A receptors appear to be absent from both nuclei. The findings suggest that release of serotonin from fibres in close proximity to trigeminal primary afferent somata could modify the transmission of action potentials from muscle spindle receptors during mastication through an action on serotonin2 receptors. In the motor nucleus and cell group k, serotonin may alter neuronal properties through actions on at least two receptor subtypes (serotonin2 and serotonin3).
Asunto(s)
Mesencéfalo/fisiología , Neuronas Motoras/fisiología , Serotonina/fisiología , Núcleos del Trigémino/fisiología , Potenciales de Acción/fisiología , Animales , Autorradiografía , Inmunohistoquímica , Maxilares/inervación , Mesencéfalo/citología , Mesencéfalo/inmunología , Husos Musculares/fisiología , Neuronas Aferentes/fisiología , Conejos , Coloración y Etiquetado , Núcleos del Trigémino/citología , Núcleos del Trigémino/inmunologíaRESUMEN
These experiments have shown that a group of neurons lateral to the trigeminal motor nucleus innervates the muscles of mastication. The work began to describe the location of digastric last-order interneurons, using the technique of transneuronal labeling with wheatgerm agglutinin-conjugated horseradish peroxide injected into the left digastric muscle of rabbits under general anaesthesia. Four to eight days later, the animals were killed with an overdose of anaesthetic and perfused. Coronal sections of the frozen brainstem were cut at 20 microns thickness and processed for peroxidase activity. Motoneurons in the ventral and caudal divisions of the trigeminal motor nucleus were labeled in all animals as expected. An additional population of neurons located ventrolaterally to the motor nucleus in cell group k were also found to be labeled if the survival time was five days or more. In an attempt to determine whether cell group k neurons were labeled transynaptically, two series of control experiments were carried out. In the first, crystals of fluorescein- and rhodamine-conjugated dextran amines and horseradish peroxidase were applied directly to central ends of cut digastric nerves. In the second, central ends of cut digastric nerves were enclosed in cuffs containing 40-60% horseradish peroxidase solutions. Again, neurons in both the trigeminal motor nucleus and cell group k were labeled suggesting that neurons within cell group k project to the digastric muscle. Similar experiments using dextran amines and wheatgerm peroxidase were carried out on the masseter muscle. Motoneurons in the dorsomedial and rostral half of the trigeminal motor nucleus, as well as primary afferent cell bodies in the mesencephalic nucleus of the trigeminal nerve, were labeled in all experiments. In addition, a population of neurons in cell group k, dorsal to those associated with the digastric muscle, were found to contain each one of the reaction products. Since it is thought that only the wheatgerm agglutinin-conjugated horseradish peroxidase transferred from one neuron to another, we conclude that cell group k neurons provide an additional innervation to the digastric and masseter muscles.
Asunto(s)
Tronco Encefálico/fisiología , Músculos Masticadores/inervación , Neuronas Motoras/fisiología , Nervios Periféricos/fisiología , Animales , Transporte Axonal , Tronco Encefálico/anatomía & histología , Peroxidasa de Rábano Silvestre , Músculo Masetero/inervación , Neuronas Motoras/citología , Músculos del Cuello/inervación , Nervios Periféricos/anatomía & histología , Conejos , Aglutinina del Germen de Trigo-Peroxidasa de Rábano Silvestre Conjugada , Aglutininas del Germen de TrigoRESUMEN
Neurons of several nuclei within the medial pontomedullar reticular formation are active during mastication, but their relationship with other elements of the pattern generating circuits have never been clearly defined. In this paper, we have studied the connection of this area with the trigeminal motor nucleus and with pools of last-order interneurons of the lateral brainstem. Retrograde tracing techniques were used in combination with immunohistochemistry to define populations of glutamatergic and GABAergic neurons. Injections of tracer into the Vth motor nucleus marked neurons in several trigeminal nuclei including the ipsilateral mesencephalic nucleus, the contralateral Vth motor nucleus, the dorsal cap of the main sensory nucleus and the rostral divisions of the spinal nucleus bilaterally. Many last-order interneurons formed a bilateral lateral band running caudally from Regio h (the zone surrounding the Vth motor nucleus), through the parvocellular reticular formation and Vth spinal caudal nucleus. Injections of tracer into Regio h, an area rich in last-order interneurons, marked, in addition to the areas listed above, a large number of neurons in the medial reticular formation bilaterally. The major difference between injection sites was that most neurons projecting to the Vth motor nucleus were located laterally, whereas most of those projecting to Regio h were found medially. Both populations contained glutamatergic and GABAergic neurons intermingled. Our results indicate that neurons of the medial reticular formation that are active during mastication influence Vth motoneurons output via relays in Regio h and other adjacent nuclei.
Asunto(s)
Tronco Encefálico/anatomía & histología , Interneuronas , Formación Reticular , Núcleos del Trigémino/anatomía & histología , Animales , Tronco Encefálico/fisiología , Interneuronas/fisiología , Masculino , Masticación/fisiología , Vías Nerviosas/anatomía & histología , Vías Nerviosas/fisiología , Conejos , Formación Reticular/fisiología , Núcleos del Trigémino/fisiologíaRESUMEN
Field EPSPs were recorded from the CA1 region of hippocampal slices under conditions in which components of the responses other than those generated by AMPA-type glutamate receptors were blocked. Laminar profile analysis indicated that the resultant potentials had separable phases: an early and fast stage followed by a late and slow stage. The location of the fast response was sensitive to stimulation position in the stratum radiatum; i.e., distal stimulation elicited maximum, negative going potentials in the distal stratum radiatum while proximal stimulation recordings were maximal in the proximal segment. The distribution of the late component was largely independent of stimulation electrode position in stratum radiatum. Current source density analysis revealed that the late response had a source in the most distal dendrites (stratum moleculare) and a sink near the cell body layer. It was not accompanied by evident changes in membrane conductance and had a decay time constant similar to the membrane time constant. Stimulation of the afferents to the basal dendrites of the pyramidal cells also resulted in a late response which again had a source in stratum moleculare. These results strongly suggest that the magnitude and waveform of the late component of the AMPA receptor mediated field potential reflects the biophysical properties of the most distal branches of the dendritic arborization. The laminar analyses also show that the late potential is minimal in the mid-stratum radiatum and thus suggest that this site is most appropriate for investigations concerned with the waveform of the fast component of AMPA receptor mediated synaptic response.
Asunto(s)
Hipocampo/fisiología , Receptores AMPA/fisiología , Animales , Estimulación Eléctrica , Electrofisiología , Masculino , Potenciales de la Membrana/fisiología , Técnicas de Cultivo de Órganos , Ratas , Ratas Sprague-Dawley , Sinapsis/fisiologíaRESUMEN
The modulatory influence of aniracetam, a drug which reversibly modifies the kinetic properties of AMPA-type glutamate receptors, on synaptic responses is reported to be detectably changed by the induction of long-term potentiation (LTP). The present study used hippocampal slices to examine three issues arising from this result. First, possible contributions of inhibitory currents and postsynaptic spiking to the aniracetam/LTP interaction were investigated with infusions of GABA receptor antagonists and topical applications of tetrodotoxin. Second, tests were carried out to determine if the altered response to aniracetam is sufficiently persistent to be a plausible substrate for the extremely stable LTP effect. Third, the nature of the change responsible for the aniracetam/LTP interaction was explored with waveform analyses and a kinetic model of the AMPA receptor. The following results were obtained. LTP reduced the effect of aniracetam on the amplitude but increased its effect on the decay time constant of field EPSPs recorded under conditions in which local spiking and inhibitory responses were blocked. The LTP-induced change in the effect of aniracetam was extremely stable in that it was still evident 75 min after induction of potentiation. Finally, the waveform distortions introduced by LTP and aniracetam could be corrected by uniform stretching of the responses, suggesting that the changes introduced by each of the manipulations are unitary in nature. These distortions and the interactions between them could be reproduced in the AMPA receptor model by representing LTP as an acceleration of channel gating kinetics.
Asunto(s)
Potenciales Postsinápticos Excitadores/efectos de los fármacos , Hipocampo/efectos de los fármacos , Potenciación a Largo Plazo/efectos de los fármacos , Nootrópicos/farmacología , Pirrolidinonas/farmacología , Receptores AMPA/efectos de los fármacos , Animales , Cinética , Modelos Neurológicos , Técnicas de Placa-Clamp , RatasRESUMEN
The catecholamine noradrenaline has been proposed to modulate the excitability of cortical neurons, and such a regulation may be mediated by specific adrenergic receptors. We characterized, using electrophysiological recordings, the types of responses of single cells in the rat visual cortex (areas 17 and 18) to the iontophoretic application of adrenergic agents. For the majority of spontaneous and visually-driven cells sampled, noradrenaline decreased the firing frequency, and in some cases of visually-driven cells could increase the signal/noise ratio. These effects were also documented after the application of the alpha-2 adrenergic agonists clonidine and oxymetazoline, and could be reduced or blocked by a previous ejection of the specific alpha-2 antagonist idazoxan. The present study supports a role for alpha-2 adrenoceptors in the modulation of sensory inputs to the visual cortex.
Asunto(s)
Agonistas alfa-Adrenérgicos/farmacología , Neuronas/fisiología , Receptores Adrenérgicos alfa/fisiología , Corteza Visual/fisiología , Antagonistas Adrenérgicos beta/farmacología , Animales , Clonidina/farmacología , Dioxanos/farmacología , Idazoxan , Masculino , Neuronas/efectos de los fármacos , Norepinefrina/farmacología , Oximetazolina/farmacología , Ratas , Ratas Endogámicas , Receptores Adrenérgicos alfa/efectos de los fármacos , Corteza Visual/efectos de los fármacosRESUMEN
In the present study we used the expression of the c-Fos-like protein as a "functional marker" to map populations of brainstem neurons involved in the generation of mastication. Experiments were conducted on urethane-anesthetized and paralyzed rabbits. In five animals (experimental group), rhythmical bouts of fictive masticatory-like motoneuron activity (cumulative duration 60-130 min) were induced by electrical stimulation of the left cortical "masticatory area" and recorded from the right digastric motoneuron pool. A control group of five animals (non-masticatory) were treated in the same way as the experimental animals with regard to surgical procedures, anesthesia, paralysis, and survival time. To detect the c-Fos-like protein, the animals were perfused, and the brainstems were cryosectioned and processed immunocytochemically. In the experimental group, the number of c-Fos-like immunoreactive neurons increased significantly in several brainstem areas. In rostral and lateral areas, increments occurred bilaterally in the borderzones surrounding the trigeminal motor nucleus (Regio h); the rostrodorsomedial half of the trigeminal main sensory nucleus; subnucleus oralis-gamma of the spinal trigeminal tract; nuclei reticularis parvocellularis pars alpha and nucleus reticularis pontis caudalis (RPc) pars alpha. Further caudally-enhanced labeling occurred bilaterally in nucleus reticularis parvocellularis and nucleus reticularis gigantocellularis (Rgc) including its pars-alpha. Our results provide a detailed anatomical record of neuronal populations that are correlated with the generation of the masticatory motor behavior.
Asunto(s)
Tronco Encefálico/metabolismo , Masticación/fisiología , Neuronas/metabolismo , Proteínas Proto-Oncogénicas c-fos/metabolismo , Anestesia , Animales , Interpretación Estadística de Datos , Vías Eferentes/citología , Vías Eferentes/fisiología , Estimulación Eléctrica , Inmunohistoquímica , Masculino , Músculos Masticadores/inervación , Músculos Masticadores/fisiología , Microelectrodos , Corteza Motora/fisiología , Neuronas Motoras/fisiología , Parálisis/fisiopatología , ConejosRESUMEN
A population of neurons in the trigeminal principal sensory nucleus (NVsnpr) fire rhythmically during fictive mastication induced in the in vivo rabbit. To elucidate whether these neurons form part of the central pattern generator (CPG) for mastication, we performed intracellular recordings in brainstem slices taken from young rats. Two cell types were defined, nonbursting (63%) and bursting (37%). In response to membrane depolarization, bursting cells, which dominated in the dorsal part of the NVsnpr, fired an initial burst followed by single spikes or recurring bursts. Non-bursting neurons, scattered throughout the nucleus, fired single action potentials. Microstimulation applied to the trigeminal motor nucleus (NVmt), the reticular border zone surrounding the NVmt, the parvocellular reticular formation or the nucleus reticularis pontis caudalis (NPontc) elicited a postsynaptic potential in 81% of the neurons tested for synaptic inputs. Responses obtained were predominately excitatory and sensitive to glutamatergic antagonists DNQX and/or APV. Some inhibitory and biphasic responses were also evoked. Bicuculline methiodide or strychnine blocked the IPSPs indicating that they were mediated by GABA(A) or glycinergic receptors. About one-third of the stimulations activated both types of neurons antidromically, mostly from the masseteric motoneuron pool of NVmt and dorsal part of NPontc. In conclusion, our new findings show that some neurons in the dorsal NVsnpr display both firing properties and axonal connections which support the hypothesis that they may participate in masticatory pattern generation. Thus, the present data provide an extended basis for further studies on the organization of the masticatory CPG network.
Asunto(s)
Potenciales de Acción/fisiología , Neuronas/clasificación , Neuronas/fisiología , Transmisión Sináptica/fisiología , Núcleos del Trigémino/citología , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/efectos de la radiación , Animales , Animales Recién Nacidos , Biotina/análogos & derivados , Biotina/metabolismo , Relación Dosis-Respuesta en la Radiación , Interacciones Farmacológicas , Estimulación Eléctrica/métodos , Potenciales Evocados/efectos de los fármacos , Potenciales Evocados/fisiología , Potenciales Evocados/efectos de la radiación , Antagonistas de Aminoácidos Excitadores/farmacología , Glicinérgicos/farmacología , Técnicas In Vitro , Neuronas/efectos de los fármacos , Neuronas/efectos de la radiación , Técnicas de Placa-Clamp/métodos , Quinoxalinas/farmacología , Ratas , Ratas Sprague-Dawley , Estricnina/farmacología , Transmisión Sináptica/efectos de los fármacos , Transmisión Sináptica/efectos de la radiaciónRESUMEN
Recent studies suggest that the nucleus pontis caudalis (nPontc) plays a role in patterning mastication through interactions with the adjacent lateral tegmentum. In this study, we used in vitro intracellular recording and staining to describe the basic membrane properties and morphology of nPontc neurones and to further explore interactions with adjacent structures, using coronal sections of the brainstem of 78 rats, aged 9-28 days. Neurones were large, with dendrites that spread in all directions, and about 64% fired tonically even in the absence of synaptic inputs. Tonic neurones were predominant in the centre of the nucleus. Electrical stimulation of all regions of the nPontc produced mixed excitatory and inhibitory effects on interneurones of lateral tegmental nuclei. Focal inactivation of the dorsal nPontc with injections of tetrodotoxin also had mixed effects on the spontaneous firing of both interneurones and motoneurones but similar injections in the ventral nPontc produced mostly increases of firing. Sixty-five percent of nPontc neurones received synaptic inputs from the lateral tegmental areas and most of these (68%) were excitatory and mediated by glutamatergic receptors. Inhibitory postsynaptic potentials were mediated by GABA(A) or glycinergic receptors. Although most responses occurred at relatively long latencies (> 2 ms), they could follow relatively high-frequency stimulation (> 50 Hz). Excitatory and inhibitory connections between ipsi- and contralateral nPontc neurones were also documented, which could contribute to bilateral coordination of jaw movements. This study provides evidence that the nPontc exerts both tonic and phasic influences on the premotor components of the masticatory central pattern generator.
Asunto(s)
Vías Nerviosas/citología , Neuronas/fisiología , Puente/citología , Tegmento Mesencefálico/anatomía & histología , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/fisiología , Potenciales de Acción/efectos de la radiación , Factores de Edad , Anestésicos Locales/farmacología , Animales , Animales Recién Nacidos , Bicuculina/farmacología , Interacciones Farmacológicas , Estimulación Eléctrica/métodos , Antagonistas de Aminoácidos Excitadores/farmacología , 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 , Lisina/metabolismo , Potenciales de la Membrana/efectos de los fármacos , Potenciales de la Membrana/fisiología , Potenciales de la Membrana/efectos de la radiación , Inhibición Neural/efectos de los fármacos , Inhibición Neural/fisiología , Inhibición Neural/efectos de la radiación , Vías Nerviosas/fisiología , Neuronas/efectos de los fármacos , Neuronas/efectos de la radiación , Puente/metabolismo , Puente/fisiología , Quinoxalinas/farmacología , Ratas , Tiempo de Reacción/fisiología , Tiempo de Reacción/efectos de la radiación , Tegmento Mesencefálico/fisiología , Tetrodotoxina/farmacologíaRESUMEN
Because of their many inputs and bilateral projections, interneurons surrounding the trigeminal motor nucleus (MotV) are thought to be very important in control of jaw movements and reflexes. However, their interactions with the trigeminal motoneurons are almost unknown. In the present study an in vitro slice preparation was used to investigate this relationship in rat. The zone bordering MotV has been subdivided into four regions: the supra-, juxta-, and intertrigeminal areas (SupV, JuxtV, and IntV, respectively) and the parvocellular reticular formation ventral and caudal to MotV. Stimulation of all areas evoked short-latency excitatory postsynaptic potentials (EPSPs) in masseteric motoneurons. Frequently the EPSPs masked inhibitory postsynaptic potentials (IPSPs) or were followed by long-lasting inhibitory potentials. Only responses obtained from stimulation of JuxtV and IntV seemed devoid of inhibitory components. The EPSPs were mediated through kainate/alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors, whereas the IPSPs appear to be due to gamma-aminobutyric acid and glycine. EPSPs and IPSPs were also recorded in SupV premotor interneurons after stimulation of IntV and MotV, respectively, thus suggesting that reciprocal connections exist between premotor areas and also between premotor interneurons of SupV and inhibitory interneurons located within MotV. It is concluded that the preparation used here will doubtless prove useful for further investigation of the circuitry involved in the bilateral coordination of the jaw.
Asunto(s)
Interneuronas/fisiología , Músculo Masetero/inervación , Neuronas Motoras/fisiología , Sinapsis/fisiología , Núcleos del Trigémino/fisiología , Animales , Bicuculina/farmacología , Carbocianinas , Estimulantes del Sistema Nervioso Central/farmacología , Estimulación Eléctrica , Antagonistas de Aminoácidos Excitadores/farmacología , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Antagonistas del GABA/farmacología , Histocitoquímica , Técnicas In Vitro , Interneuronas/efectos de los fármacos , Músculo Masetero/efectos de los fármacos , Músculo Masetero/fisiología , Neuronas Motoras/efectos de los fármacos , Quinoxalinas/farmacología , Ratas , Estricnina/farmacología , Sinapsis/efectos de los fármacos , Núcleos del Trigémino/citología , Núcleos del Trigémino/efectos de los fármacosRESUMEN
Numerous evidence suggests that interneurons located in the lateral tegmentum at the level of the trigeminal motor nucleus contribute importantly to the circuitry involved in mastication. However, the question of whether these neurons participate actively to genesis of the rhythmic motor pattern or simply relay it to trigeminal motoneurons remains open. To answer this question, intracellular recordings were performed in an in vitro slice preparation comprising interneurons of the peritrigeminal area (PeriV) surrounding the trigeminal motor nucleus (NVmt) and the parvocellular reticular formation ventral and caudal to it (PCRt). Intracellular and extracellular injections of anterograde tracers were also used to examine the local connections established by these neurons. In 97% of recordings, electrical stimulation of adjacent areas evoked a postsynaptic potential (PSP). These PSPs were primarily excitatory, but inhibitory and biphasic responses were also induced. Most occurred at latencies longer than those required for monosynaptic transmission and were considered to involve oligosynaptic pathways. Both the anatomical and physiological findings show that all divisions of PeriV and PCRt are extensively interconnected. Most responses followed high-frequency stimulation (50 Hz) and showed little variability in latency indicating that the network reliably distributes inputs across all areas. In all neurons but one, excitatory postsynaptic potentials (EPSPs) or inhibitory postsynaptic potentials (IPSPs) were also elicited by stimulation of NVmt, suggesting the existence of excitatory and inhibitory interneurons within the motor nucleus. In a number of cases, these PSPs were reproduced by local injection of glutamate in lieu of the electrical stimulation. All EPSPs induced by stimulation of PeriV, PCRt, or NVmt were sensitive to ionotropic glutamate receptor antagonists 6-cyano-7-dinitroquinoxaline and D,L-2-amino-5-phosphonovaleric acid, while IPSPs were blocked by bicuculline and strychnine, antagonists of GABA(A) and glycine receptors. Examination of PeriV and PCRt intrinsic properties indicate that they form a fairly uniform network. Three types of neurons were identified on the basis of their firing adaptation properties. These types were not associated with particular regions. Only 5% of all neurons showed bursting behavior. Our results do not support the hypothesis that neurons of PeriV and PCRt participate actively to rhythm generation, but suggest instead that they are driven by rhythmical synaptic inputs. The organization of the network allows for rapid distribution of this rhythmic input across premotoneuron groups.
Asunto(s)
Interneuronas/fisiología , Puente/fisiología , Formación Reticular/fisiología , Núcleos del Trigémino/fisiología , Vías Aferentes/fisiología , Animales , Vías Eferentes/fisiología , Estimulación Eléctrica , Electrofisiología , Antagonistas de Aminoácidos Excitadores/farmacología , Potenciales Postsinápticos Excitadores/efectos de los fármacos , Potenciales Postsinápticos Excitadores/fisiología , Ácido Glutámico/farmacología , Técnicas In Vitro , Interneuronas/efectos de los fármacos , Inhibición Neural/fisiología , Puente/citología , Ratas , Tiempo de Reacción/fisiología , Formación Reticular/citología , Transmisión Sináptica/efectos de los fármacos , Núcleos del Trigémino/citologíaRESUMEN
The catecholamines noradrenaline and dopamine have been proposed as neuromodulators of cortical neuron excitability, and such a regulation could be mediated by specific adrenergic and dopaminergic receptors. We characterized electrophysiologically some of the types of responses to the iontophoretic application of adrenergic and dopaminergic agonists and antagonists on single cells in the rat visual cortex (areas occipital 1 monocular or Oc1M and occipital 1 binocular or Oc1B). For the majority of spontaneously active and visual cortical cells, noradrenaline and dopamine decreased the firing frequency. In the case of visually driven (synaptically activated) neurons, background firing was the main component of the response to be inhibited by the administration of noradrenaline, clonidine, and oxymetazoline, leading to an enhancement of the signal-to-noise ratio. Since these effects could be reduced or blocked by a previous ejection of the specific alpha 2-antagonist idazoxan, the findings support a role for alpha 2-adrenergic receptors in the transmission of sensory inputs to the visual cortex. These effects were not found with the mixed alpha-adrenergic agonist phenylephrine nor with the beta-agonist isoproterenol. Finally, the use of the inhibitory amino acid GABA rules out a simple hyperpolarizing response as the mechanism underlying noradrenaline modulatory effects in the cerebral cortex.