RESUMO
We hypothesised that pentobarbital would improve upper airway mechanics based on an increase in latency to arousal and amplitude of the phasic genioglossus electromyogram (EMG), and a decrease in the active upper airway critical closing pressure (P(crit)). 12 healthy subjects received pentobarbital (100 mg) or placebo in a double-blind, crossover protocol. During wakefulness, we measured the genioglossus reflex response to negative pressure pulses. During sleep, carbon dioxide was insufflated into the inspired air. Airway pressure was then decreased in a stepwise fashion until arousal from sleep. With basal breathing during sleep: flow rate was lower in volunteers given pentobarbital; end-tidal CO(2) concentration and upper airway resistance were greater; and P(crit) was unaffected (pentobarbital mean ± SD -11.7 ± 4.5 versus placebo -10.25 ± 3.6 cmH(2)O; p = 0.11). Pentobarbital increased the time to arousal (297 ± 63s versus 232 ± 67 s; p<0.05), at which time phasic genioglossus EMG was higher (6.2 ± 4.8% maximal versus 3.1 ± 3%; p<0.05) as were CO(2) levels. The increase in genioglossus EMG after CO(2) administration was greater after pentobarbital versus placebo. Pentobarbital did not affect the genioglossus negative-pressure reflex. Pentobarbital increases the time to arousal and stimulates genioglossus muscle activity, but it also increases upper airway resistance during sleep.
Assuntos
Pentobarbital/farmacologia , Adolescente , Adulto , Estudos Cross-Over , Método Duplo-Cego , Eletromiografia/métodos , Feminino , Humanos , Hipnóticos e Sedativos/farmacologia , Pulmão/patologia , Masculino , Pessoa de Meia-Idade , Placebos , Respiração , Transtornos do Sono-VigíliaRESUMO
BACKGROUND: Cholinesterase inhibitor-based reversal agents, given in the absence of neuromuscular block, evoke a partial upper airway obstruction by decreasing skeletal upper airway muscle function. Sugammadex reverses neuromuscular block by encapsulating rocuronium. However, its effects on upper airway integrity and breathing are unknown. METHODS: Fifty-one adult male rats were anaesthetized with isoflurane, tracheostomized, and a femoral artery and vein were cannulated. First, we compared the efficacy of sugammadex 15 mg kg(-1) and neostigmine 0.06 mg kg(-1) to reverse respiratory effects of rocuronium-induced partial paralysis [train-of-four ratio (T4/T1)=0.5]. Subsequently, we compared the safety of sugammadex and neostigmine given after recovery of the T4/T1 to 1, by measuring phasic genioglossus activity and breathing. RESULTS: During partial paralysis (T4/T1=0.5), time to recovery of minute volume to baseline values was 10.9 (2), 75.8 (18), and 153 (54) s with sugammadex, neostigmine, and placebo, respectively (sugammadex was significantly faster than neostigmine and placebo, P<0.05). Recovery of T4/T1 was also faster for sugammadex than neostigmine and placebo. Neostigmine administration after complete recovery of T4/T1 decreased upper airway dilator muscle activity to 64 (30)% of baseline and decreased tidal volume (P<0.05 for both variables), whereas sugammadex had no effect on either variable. CONCLUSIONS: In contrast to neostigmine, which significantly impairs upper airway dilator muscle activity when given after recovery from neuromuscular block, a reversal dose of sugammadex given under the same conditions does not affect genioglossus muscle activity and normal breathing. Human studies will be required to evaluate the clinical relevance of our findings.
Assuntos
Inibidores da Colinesterase/farmacologia , Neostigmina/farmacologia , Respiração/efeitos dos fármacos , Músculos Respiratórios/efeitos dos fármacos , gama-Ciclodextrinas/farmacologia , Androstanóis/antagonistas & inibidores , Androstanóis/farmacologia , Período de Recuperação da Anestesia , Anestésicos Inalatórios , Animais , Eletromiografia/efeitos dos fármacos , Isoflurano , Masculino , Bloqueio Neuromuscular , Fármacos Neuromusculares não Despolarizantes/antagonistas & inibidores , Fármacos Neuromusculares não Despolarizantes/farmacologia , Ratos , Ratos Sprague-Dawley , Músculos Respiratórios/fisiologia , Rocurônio , SugammadexRESUMO
We have cloned from rat brain the cDNA encoding an 89,828-Da kinesin-related polypeptide KIF3C that is enriched in brain, retina, and lung. Immunocytochemistry of hippocampal neurons in culture shows that KIF3C is localized to cell bodies, dendrites, and, in lesser amounts, to axons. In subcellular fractionation experiments, KIF3C cofractionates with a distinct population of membrane vesicles. Native KIF3C binds to microtubules in a kinesin-like, nucleotide-dependent manner. KIF3C is most similar to mouse KIF3B and KIF3A, two closely related kinesins that are normally present as a heteromer. In sucrose density gradients, KIF3C sediments at two distinct densities, suggesting that it may be part of two different multimolecular complexes. Immunoprecipitation experiments show that KIF3C is in part associated with KIF3A, but not with KIF3B. Unlike KIF3B, a significant portion of KIF3C is not associated with KIF3A. Consistent with these biochemical properties, the distribution of KIF3C in the CNS has both similarities and differences compared with KIF3A and KIF3B. These results suggest that KIF3C is a vesicle-associated motor that functions both independently and in association with KIF3A.
Assuntos
Cinesinas/metabolismo , Sequência de Aminoácidos , Animais , Encéfalo/metabolismo , Movimento Celular , Clonagem Molecular , Imuno-Histoquímica , Cinesinas/química , Cinesinas/genética , Masculino , Camundongos , Dados de Sequência Molecular , Neurônios/metabolismo , Conformação Proteica , Ratos , Ratos Sprague-Dawley , Frações Subcelulares/metabolismo , Distribuição TecidualRESUMO
Adenosine has been proposed as a homeostatic "sleep factor" that promotes the transition from waking to sleep by affecting several sleep-wake regulatory systems. In the basal forebrain, adenosine accumulates during wakefulness and, when locally applied, suppresses neuronal activity and promotes sleep. However, the neuronal phenotype mediating these effects is unknown. We used whole-cell patch-clamp recordings in in vitro rat brain slices to investigate the effect of adenosine on identified cholinergic and noncholinergic neurons of the magnocellular preoptic nucleus and substantia innominata. Adenosine (0.5-100 microM) reduced the magnocellular preoptic nucleus and substantia innominata cholinergic neuronal firing rate by activating an inwardly rectifying potassium current that reversed at -82 mV and was blocked by barium (100 microM). Application of the A1 receptor antagonist 8-cyclo-pentyl-theophylline (200 nM) blocked the effects of adenosine. Adenosine was also tested on two groups of electrophysiologically distinct noncholinergic magnocellular preoptic nucleus and substantia innominata neurons. In the first group adenosine, via activation of postsynaptic A1 receptors, reduced spontaneous firing via inhibition of the hyperpolarization-activated cation current. Blocking the H-current with ZD7288 (20 microM) abolished adenosine effects on these neurons. The second group was not affected by adenosine. These results demonstrate that, in the magnocellular preoptic nucleus and substantia innominata region of the basal forebrain, adenosine inhibits both cholinergic neurons and a subset of noncholinergic neurons. Both of these effects occur via postsynaptic A1 receptors, but are mediated downstream by two separate mechanisms.
Assuntos
Acetilcolina/metabolismo , Adenosina/metabolismo , Fibras Colinérgicas/metabolismo , Neurônios/metabolismo , Área Pré-Óptica/metabolismo , Substância Inominada/metabolismo , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Adenosina/farmacologia , Agonistas do Receptor A1 de Adenosina , Antagonistas do Receptor A1 de Adenosina , Animais , Fibras Colinérgicas/efeitos dos fármacos , Feminino , Masculino , Inibição Neural/efeitos dos fármacos , Inibição Neural/fisiologia , Neurônios/efeitos dos fármacos , Técnicas de Cultura de Órgãos , Técnicas de Patch-Clamp , Canais de Potássio/efeitos dos fármacos , Canais de Potássio/metabolismo , Área Pré-Óptica/citologia , Área Pré-Óptica/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Receptor A1 de Adenosina/metabolismo , Sono/efeitos dos fármacos , Sono/fisiologia , Substância Inominada/citologia , Substância Inominada/efeitos dos fármacos , Transmissão Sináptica/efeitos dos fármacos , Transmissão Sináptica/fisiologia , Canais de Potencial de Receptor Transitório/efeitos dos fármacos , Canais de Potencial de Receptor Transitório/metabolismoRESUMO
We examined the functional organization of the parabrachial complex (PB) by mapping the cardiovascular and respiratory responses to PB microstimulation in anesthetized rats. The PB was explored with 100 microns resolution, at threshold doses of electrical current (5 microA) and glutamate (10-500 pmols), and the locations of stimulation sites were identified by small iontophoretic or pressure injections of biocytin or Phaseolus vulgaris leucoagglutinin. Threshold doses of either L-glutamate or electrical current pulses caused pressor-tachycardic responses that mapped to the outer edge of the external lateral subnucleus while depressor bradycardic responses were elicited from stimuli near the dorsal lateral subnucleus. Pressor responses persisted in paralyzed, ventilated animals and were thus not dependent upon concomitant respiratory changes. Cardiac arrhythmias sometimes occurred during large pressor responses and during augmented breaths that occurred during or following PB stimulation. These observations indicate that the PB contains at least two distinct neuronal systems that are potently and opposingly involved in cardiovascular control. The locations of the sites giving the most potent responses implicate specific ascending and descending pathways as substrates for the cardiovascular responses.
Assuntos
Mapeamento Encefálico/métodos , Sistema Cardiovascular/efeitos dos fármacos , Glutamatos/farmacologia , Animais , Arritmias Cardíacas/induzido quimicamente , Pressão Sanguínea/efeitos dos fármacos , Pressão Sanguínea/fisiologia , Fenômenos Fisiológicos Cardiovasculares , Estimulação Elétrica , Corantes Fluorescentes , Ácido Glutâmico , Frequência Cardíaca/efeitos dos fármacos , Frequência Cardíaca/fisiologia , Masculino , Microeletrodos , Microinjeções , Ratos , Ratos Sprague-Dawley , Respiração/efeitos dos fármacos , Respiração/fisiologiaRESUMO
The parabrachial complex is made up of at least 11 cytoarchitectonically distinct subnuclei which differ in their anatomical connections and neurotransmitter content, as well as the functions they subserve. To determine whether parabrachial subnuclei also express different types of glutamate receptors, we undertook a light microscopic examination of the regional distribution of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptor subunits within the parabrachial complex using antibodies directed against synthetic peptides corresponding to the C-terminal parts of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptor subunits. Antibodies that recognize GluR1 subunits stained cells mainly in the central lateral parabrachial subnucleus, whereas GluR4 antibodies selectively stained cells in the internal lateral subnucleus. In contrast, antibodies directed against the GLuR2/3 subunits stained neurons in every parabrachial subnucleus, although the most dense labelling was seen in the external lateral cell group. These differences in expression of alpha-amino-3-hydroxy-5-methyl-4- isoxazolepropionate receptor subtypes may eventually allow selective activation or inhibition of specific subsets of neurons in the parabrachial complex.
Assuntos
Núcleos Cerebelares/metabolismo , Ponte/metabolismo , Receptores de AMPA/metabolismo , Animais , Núcleos Cerebelares/citologia , Eletrofisiologia , Aminoácidos Excitatórios/farmacologia , Imuno-Histoquímica , Masculino , Neurônios/metabolismo , Fenótipo , Ponte/citologia , Ratos , Ratos Sprague-DawleyRESUMO
Hypoglossal motoneurons (HMNs), which innervate the tongue muscles, are involved in several important physiological functions, including the maintenance of upper airway patency. The neural mechanisms that affect HMN excitability are therefore important determinants of effective breathing. Obstructive sleep apnea is a disorder characterized by recurrent collapse of the upper airway that is likely due to decline of pharyngeal motoneuron activity during sleep. Because cholinergic neuronal activity is closely coupled to wake and sleep states, we tested the effects and pharmacology of nicotinic acetylcholine receptor (nAChR) activation on HMNs. We made intracellular recordings from HMNs in medullary slices from neonatal rats and found that local application of the nicotinic agonist, 1,1-dimethyl-4-phenylpiperazinium iodide, excited HMNs by a Ca(2+)-sensitive, and TTX-insensitive inward current that was blocked by dihydro-beta-erythroidine (IC(50): 19+/-3 nM), methyllycaconitine (IC(50): 32+/-7 nM), and mecamylamine (IC(50): 88+/-11 nM), but not by alpha-bungarotoxin (10 nM). This is consistent with responses being mediated by postsynaptic nAChRs that do not contain the alpha7 subunit. These results suggest that nAChR activation may contribute to central maintenance of upper airway patency and that the decline in firing rate of cholinergic neurons during sleep could potentially disfacilitate airway dilator muscle activity, contributing to airway obstruction.
Assuntos
Nervo Hipoglosso/metabolismo , Lisina/análogos & derivados , Bulbo/metabolismo , Neurônios Motores/metabolismo , Receptores Nicotínicos/metabolismo , Fenômenos Fisiológicos Respiratórios/efeitos dos fármacos , Apneia Obstrutiva do Sono/metabolismo , Sono/fisiologia , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Animais , Animais Recém-Nascidos , Agonistas Colinérgicos/farmacologia , Iodeto de Dimetilfenilpiperazina/farmacologia , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Potenciais Pós-Sinápticos Excitadores/fisiologia , Nervo Hipoglosso/efeitos dos fármacos , Bulbo/citologia , Bulbo/efeitos dos fármacos , Neurônios Motores/efeitos dos fármacos , Vias Neurais/efeitos dos fármacos , Vias Neurais/fisiologia , Agonistas Nicotínicos/farmacologia , Ratos , Ratos Sprague-Dawley , Receptores Nicotínicos/efeitos dos fármacos , Apneia Obstrutiva do Sono/fisiopatologia , Transmissão Sináptica/efeitos dos fármacos , Transmissão Sináptica/fisiologia , Tetrodotoxina/farmacologia , Receptor Nicotínico de Acetilcolina alfa7RESUMO
The ventrolateral preoptic nucleus (VLPO) is a key regulator of behavioral state that promotes sleep by directly inhibiting brain regions that maintain wakefulness. Subarachnoid administration of adenosine (AD) or AD agonists promotes sleep and induces expression of Fos protein in VLPO neurons. Therefore, activation of VLPO neurons may contribute to the somnogenic actions of AD. To define the mechanism through which AD activates VLPO neurons, we prepared hypothalamic slices from 9 to 12-day-old rat pups and recorded from 43 neurons in the galaninergic VLPO cluster; nine neurons contained galanin mRNA by post hoc in situ hybridization. Bath application of AD (20 microM) to seven of these neurons had no direct effect but caused a significant decrease in the frequency of spontaneous miniature inhibitory postsynaptic currents in the presence of tetrodotoxin, indicating a presynaptic site of action. We conclude that AD-mediated disinhibition increases the excitability of VLPO neurons thus contributing to the somnogenic properties of AD.
Assuntos
Adenosina/metabolismo , Vias Aferentes/metabolismo , Área Pré-Óptica/metabolismo , Terminações Pré-Sinápticas/metabolismo , Transmissão Sináptica/fisiologia , Ácido gama-Aminobutírico/metabolismo , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Adenosina/farmacologia , Vias Aferentes/citologia , Vias Aferentes/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Feminino , Galanina/genética , Masculino , Inibição Neural/efeitos dos fármacos , Inibição Neural/fisiologia , Técnicas de Cultura de Órgãos , Área Pré-Óptica/citologia , Área Pré-Óptica/efeitos dos fármacos , Terminações Pré-Sinápticas/efeitos dos fármacos , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Sono/efeitos dos fármacos , Sono/fisiologia , Transmissão Sináptica/efeitos dos fármacosRESUMO
The inhibitory and excitatory amino acid neurotransmitter receptors in the mammalian central nervous system mediate functionally opposite synaptic responses yet appear to share certain structural features. Recent conceptual advances in this field have relied heavily on information obtained by single channel analyses, by the expression of receptors in oocytes, and by autoradiographic studies of receptor distribution among brain receptors. This article reviews the pharmacology, cellular physiology, and regional distribution of these receptors and discusses their role in several well-characterized neurological disease states. Also reviewed are the recent advances made in purifying (in some instances cloning) the receptors and uptake sites involved in synaptic transmission in the brain. Throughout, the emphasis is on synthesis and concept rather than on methodological detail.
Assuntos
Sistema Nervoso Central/metabolismo , Receptores de Superfície Celular , Animais , Sistema Nervoso Central/fisiologia , Sistema Nervoso Central/fisiopatologia , Humanos , Canais Iônicos/efeitos dos fármacos , Canais Iônicos/fisiologia , Doenças do Sistema Nervoso/metabolismo , Inibição Neural , Receptores de Aminoácido , Receptores de Superfície Celular/efeitos dos fármacos , Receptores de Superfície Celular/metabolismo , Receptores de Superfície Celular/fisiologiaRESUMO
Intracellular recordings from CA3b/c neurons in rat hippocampal slices showed that reduction of the extracellular Cl- concentration from 136 to 53 mM produced a positive (+10 mV) shift in the reversal potential of GABAergic inhibitory postsynaptic potentials (IPSPs). This shift was not significantly different from the shift produced by raising K+ from 3.5 to 8.5 mM. Spontaneous interictal bursting occurred in both low Cl- and high K+. Extracellular recordings from the pyramidal cell layer in the CA3b/c region of hippocampal slices showed that bursts in 56 mM Cl- were of the same waveform and intensity as bursts produced by high K+. However the frequency of spontaneous bursting was much lower (6.6 +/- 1.2/min, n = 10) in low Cl- compared to high K+ (42.2 +/- 3.0/min, n = 33). Burst frequency was a linear function of the shift in IPSP reversal potential produced by high K+, but not low Cl-. Replacing 60% of the Cl- with methylsulfate or isethionate was sufficient to produce spontaneous bursting, whereas it was necessary to replace 80% of the Cl- when propionate was used as a substitute. All 3 Cl- substitutes lowered the ionized Ca2+ concentration, but raising the extracellular Ca2+ concentration back to normal did not change the burst frequency. Since the amplitude of IPSPs is reduced to a similar extent in low Cl- and high K+ solutions, whereas bursting is much faster in high K+, we suggest that impaired GABAergic inhibition is insufficient to fully account for spontaneous interictal bursting that is produced in hippocampal slices by raised extracellular K+.
Assuntos
Cloretos/farmacologia , Epilepsia/fisiopatologia , Hipotálamo/fisiologia , Neurônios/fisiologia , Potássio/farmacologia , Ácido gama-Aminobutírico/fisiologia , Animais , Hipotálamo/efeitos dos fármacos , Hipotálamo/fisiopatologia , Técnicas In Vitro , Potenciais da Membrana/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Tratos Piramidais/efeitos dos fármacos , Tratos Piramidais/fisiologia , Ratos , Sinapses/efeitos dos fármacos , Sinapses/fisiologiaRESUMO
Spontaneous interictal bursts occurred at a high rate in rat hippocampal slices bathed in raised [K]o (7-10.5 mM) and at a slower rate in slices bathed in low [Cl]o (53 mM) solutions. Intracellular recordings from CA3b/c neurons bathed in high [K]o showed that the amplitude of the afterhyperpolarization that followed each spontaneous epileptiform burst varied -50 +/- 7 mV per decade change in [K]o (n = 7). Afterhyperpolarization duration was highly correlated with the interburst interval in 7-10.5 mM [K]o but not in low [Cl]o (3.5 mM [K]o). The duration of the afterhyperpolarization accounted for 79 +/- 2% of the interburst interval (1.4 +/- 0.1 s) in 8.5 mM [K]o but only 51 +/- 8% in low [Cl]o (interval 6.6 +/- 1.2 s). Treatment of slices bathed in 8.5 mM [K]o with phorbol 12,13-diacetate (2-20 microM) caused a decrease in the amplitude and duration of the afterhyperpolarization and a concomitant 41% increase in burst rate. In addition, spontaneous discharges that resembled the tonic phase of electrographic seizures appeared in the CA3 region. The GABAB receptor blocker phaclofen (0.2-1 mM) had no effect on the burst rate. We suggest that the epileptiform burst afterhyperpolarization limits the maximum rate of spontaneous interictal-like bursting and that a decrease in its duration allowed the subsequent expression of electrographic seizure activity in area CA3.
Assuntos
Epilepsia/fisiopatologia , Hipocampo/fisiopatologia , Potenciais de Ação/efeitos dos fármacos , Animais , Baclofeno/análogos & derivados , Baclofeno/farmacologia , Hipocampo/efeitos dos fármacos , Técnicas In Vitro , Potenciais da Membrana/efeitos dos fármacos , Ésteres de Forbol/farmacologia , Potássio/fisiologia , RatosRESUMO
The parabrachial nucleus (PB) is a major relay of noxious and non-noxious visceral sensory information from the nucleus of the solitary tract, spinal cord, and spinal trigeminal nucleus to the forebrain. The nucleus of the solitary tract, spinal cord, and trigeminal dorsal horns contain many enkephalin- and dynorphin-immunoreactive neurons that project to the PB. To study the role of mu-opioid receptors in relaying these inputs, we examined the distribution of mu-opioid receptor immunoreactivity in the PB. The most intense staining was in the external lateral parabrachial subnucleus (PBel), including dendrites extending from the PBel into the lateral crescent subnucleus. Because the Pbel is a major source of projections to the amygdala, we combined retrograde tracing from the central nucleus of the amygdala with immunohistochemistry for mu-opioid receptors. These experiments showed that mu-opioid receptors are expressed by Pbel neurons that project to the amygdala, including those Pbel neurons whose dendrites extend into the lateral crescent subnucleus. These results indicate that mu-opioid receptors in the PB may mediate or modulate nociceptive information relayed to the amygdala from medullary or spinal cord neurons that terminate not only in the Pbel, but also in the adjacent lateral crescent parabrachial subnucleus.
Assuntos
Tonsila do Cerebelo/citologia , Neurônios/química , Ponte/citologia , Receptores Opioides mu/análise , Animais , Anticorpos , Pressão Sanguínea , Dendritos/química , Imunofluorescência , Masculino , Vias Neurais , Neurônios/ultraestrutura , Dor/fisiopatologia , Ratos , Receptores Opioides mu/imunologiaRESUMO
We used a recombinant adeno-associated virus vector (AAV) to deliver a foreign gene, green fluorescent protein (GFP), into mature neurons in adult rat CNS in vivo. Microinjections of AAV as small as 50 nl transduced hundreds of neurons at the injection site. There was virtually no retrograde transport as fewer than one neuron per brain was found distant from the injection site that exhibited GFP immunoreactivity. The gene product, GFP, filled the entire neuronal cytoplasmic compartment; GFP immunoreactivity was robust in cell bodies, axons, and nerve terminals. There was no tissue damage at the injection sites or pathogenicity indicated by changes in astrocytic or microglial markers. There was no inflammatory response as judged by leukocytic invasion. Gene expression in transduced cells was robust and apparently permanent: there was no evidence of phenotypic reversion up to 12 weeks following infection. AAV is an excellent vector for introducing foreign genes into mature CNS neurons. Not only might it be an ideal vehicle for gene therapy, but also the GFP-containing AAV presents a new strategy for tracing long axonal pathways in the CNS, which is difficult with current tracers (PHAL, biotinylated dextrans).
Assuntos
Transporte Axonal/fisiologia , Encéfalo/fisiologia , Dependovirus/genética , Expressão Gênica , Vetores Genéticos/farmacologia , Vias Neurais/fisiologia , Transgenes/genética , Animais , Transporte Axonal/genética , Linhagem Celular , Genes Reporter , Proteínas de Fluorescência Verde , Humanos , Proteínas Luminescentes/análise , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Recombinação GenéticaRESUMO
The parabrachial complex, also known as the pneumotaxic center or pontine respiratory group, has long been recognized as an important participant in respiratory control. One line of evidence supporting this idea is the demonstration of changes in breathing pattern following injection of neuroactive substances into or near the parabrachial complex. However, it is not yet known exactly which cell groups and projections mediate those responses. In order to address this issue, we explored the topographic organization of respiratory responses to chemical stimulation of the parabrachial complex of the rat and examined the descending projections of the most sensitive sites. Injection of glutamate (5-100 pmol) at specific sites in or near the parabrachial nucleus produced three distinct site-specific response patterns. First, hyperpnea followed glutamate injection into far rostral and midcaudal areas of the Kölliker-Fuse nucleus and most of the lateral parabrachial nucleus, including the external lateral, central lateral, dorsal lateral, and superior lateral subnuclei. Threshold hyperpneic effects were manifested as single, deepened breaths of premature onset. Suprathreshold doses of glutamate at these locations produced tachypnea. Neurons in these sites projected to the ventral respiratory group in the ventrolateral medulla. Second, the most intense inspiratory facilitatory responses were seen at mid to rostral levels of the Kölliker-Fuse nucleus, near the ventrolateral tip of the superior cerebellar peduncle. Even at threshold doses of glutamate, exhalation was incomplete, resulting in a breathing pattern that resembled apneusis (an inspiratory cramp). This site contained an especially dense cluster of neurons that projected either to the ventrolateral medulla or to the dorsal respiratory group in the nucleus of the solitary tract, but not to both areas. The third type of response, decreases in respiratory rate, occurred following glutamate injection at the most lateral and ventral boundaries of the Kölliker-Fuse nucleus. The most sensitive apneic sites were not found in the parabrachial nucleus but along the dorsal and medial edge of the principal sensory trigeminal nucleus and extending ventrally between the sensory and motor trigeminal nuclei. Scattered neurons in these sites were retrogradely labeled from the ventral but not the dorsal respiratory group. These results indicate that there are anatomically and functionally distinct cell populations in and near the parabrachial complex that, when chemically stimulated, can produce specific and sometimes opposing effects on respiration. The predominant effect of lateral parabrachial stimulation is respiratory facilitation, while inhibitory effects are elicited by trigeminal injections of glutamate.
Assuntos
Ácido Glutâmico/farmacologia , Ponte/fisiologia , Respiração/fisiologia , Animais , Pressão Sanguínea/efeitos dos fármacos , Mapeamento Encefálico , Injeções , Masculino , Bulbo/citologia , Bulbo/fisiologia , Ponte/citologia , Ponte/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Respiração/efeitos dos fármacosRESUMO
Apnea is an important protective response to upper airway irritation, but the central mechanisms responsible for eliciting sensory-induced apnea are not well understood. Recent studies have emphasized the Kölliker-Fuse nucleus in producing apnea and proposed a trigeminoparabrachial pathway for mediating these reflexes. However, in our earlier study of apneic responses produced by glutamate stimulation in the dorsolateral pons, we found that apnea was elicited from the area just ventral to the Kölliker-Fuse nucleus, rather than within it. Because this region was not known to be involved in respiratory control, we combined chemical microstimulation with both anterograde and retrograde axonal tracing to characterize the sites in the pons that produce apneic responses. We found that apneic sites were consistently associated with the intertrigeminal region, between the principal sensory and motor trigeminal nuclei. Injections of anterograde tracer at these sites labeled terminals in the ventral respiratory group, in the ventrolateral medulla. Injection of retrograde tracer into this target region in the ventrolateral medulla disclosed a previously unrecognized population of neurons among the trigeminal motor rootlets. Injection of retrograde tracer into this intertrigeminal region demonstrated inputs from portions of the spinal trigeminal nucleus and the nucleus of the solitary tract that have been associated with producing sensory apnea. Our observations suggest that the intertrigeminal region receives a convergence of sensory inputs capable of driving apneic responses and that it may represent a common link between input from different portions of the airway and the respiratory neurons that mediate apneic reflexes.
Assuntos
Tronco Encefálico/fisiologia , Rede Nervosa , Reflexo Anormal/fisiologia , Síndromes da Apneia do Sono/fisiopatologia , Núcleos do Trigêmeo/fisiologia , Animais , Toxina da Cólera/farmacologia , Ácido Glutâmico/farmacologia , Masculino , Bulbo/fisiopatologia , Ponte/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Centro Respiratório/fisiopatologia , Estimulação Química , Conjugado Aglutinina do Germe de Trigo-Peroxidase do Rábano SilvestreRESUMO
1. Spontaneous discharges that resemble interictal spikes arise in area CA3 b/c of rat hippocampal slices bathed in 8.5 mM [K+]o. Excitatory postsynaptic potentials (EPSPs) also appear at irregular intervals in these cells. The role of local synaptic excitation in burst initiation was examined with intracellular and extracellular recordings from CA3 pyramidal neurons. 2. Most (70%) EPSPs were small (less than 2 mV in amplitude), suggesting that they were the product of quantal release or were evoked by a single presynaptic action potential in another cell. It is unlikely that most EPSPs were evoked by a presynaptic burst of action potentials. Indeed, intrinsic burst firing was not prominent in CA3 b/c pyramidal cells perfused in 8.5 mM [K+]o. 3. The likelihood of occurrence and the amplitude of EPSPs were higher in the 50-ms interval just before the onset of each burst than during a similar interval 250 ms before the burst. This likely reflects increased firing probability of CA3 neurons as they emerge from the afterhyperpolarization (AHP) and conductance shunt associated with the previous burst. 4. Perfusion with 2 microM 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), a potent quisqualate receptor antagonist, decreased the frequency of EPSPs in CA3 b/c neurons from 3.6 +/- 0.9 to 0.9 +/- 0.3 (SE) Hz. Likewise, CNQX reversibly reduced the amplitude of evoked EPSPs in CA3 b/c cells. 5. Spontaneous burst firing in 8.5 mM [K+]o was abolished in 11 of 31 slices perfused with 2 microM CNQX.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Hipocampo/fisiologia , Neurônios/fisiologia , Potássio/farmacologia , 6-Ciano-7-nitroquinoxalina-2,3-diona , Animais , Eletrofisiologia , Hipocampo/citologia , Masculino , Quinoxalinas/farmacologia , Ratos , Ratos Endogâmicos , Tempo de ReaçãoRESUMO
Reduction of the threshold of cardiovascular and neuronal responses to facial and intracranial stimulation reflects central sensitization and cutaneous allodynia in a rat model of migraine. Current theories propose that migraine pain is caused by chemical activation of meningeal perivascular fibers. We previously found that chemical irritation of the dura causes trigeminovascular fibers innervating the dura and central trigeminal neurons receiving convergent input from the dura and skin to respond to low-intensity mechanical and thermal stimuli that previously induced minimal or no responses. One conclusion of these studies was that when low- and high-intensity stimuli induce responses of similar magnitude in nociceptive neurons, low-intensity stimuli must be as painful as the high-intensity stimuli. The present study investigates in anesthetized rats the significance of the changes in the responses of central trigeminal neurons (i.e., in nucleus caudalis) by correlating them with the occurrence and type of the simultaneously recorded cardiovascular responses. Before chemical stimulation of the dura, simultaneous increases in neuronal firing rates and blood pressure were induced by dural indentation with forces >/= 2.35 g and by noxious cutaneous stimuli such as pinching the skin and warming > 46 degrees C. After chemical stimulation, similar neuronal responses and blood pressure increases were evoked by much smaller forces for dural indentation and by innocuous cutaneous stimuli such as brushing the skin and warming it to >/= 43 degrees C. The onsets of neuronal responses preceded the onsets of depressor responses by 1.7 s and pressor responses by 4.0 s. The duration of neuronal responses was 15 s, whereas the duration of depressor responses was shorter (5.8 s) and pressor responses longer (22.7 s) than the neuronal responses. We conclude that the facilitated cardiovascular and central trigeminal neuronal responses to innocuous stimulation of the skin indicate that when dural stimulation induces central sensitization, innocuous stimuli are as nociceptive as noxious stimuli had been before dural stimulation and that a similar process might occur during the development of cutaneous allodynia during migraine.
Assuntos
Sistema Cardiovascular/fisiopatologia , Cabeça/fisiologia , Hiperalgesia/fisiopatologia , Transtornos de Enxaqueca/fisiopatologia , Neurônios/fisiologia , Animais , Pressão Sanguínea/fisiologia , Sistema Cardiovascular/inervação , Modelos Animais de Doenças , Dura-Máter/fisiologia , Masculino , Medição da Dor , Estimulação Física , Ratos , Ratos Sprague-Dawley , Pele/inervação , Estimulação Química , Fatores de Tempo , Núcleo Inferior Caudal do Nervo Trigêmeo/fisiologia , Nervo Trigêmeo/fisiologiaRESUMO
Intracellular and extracellular recordings were made from pyramidal neurons in hippocampal slices in order to study spontaneous paroxysmal bursting induced by raising the extracellular potassium concentration from 3.5 to 8.5 mM. Extracellular recordings from all hippocampal subfields indicated that spontaneous bursts appeared to originate in region CA3c or CA3b as judged by burst onset. Burst intensity was also greatest in regions CA3b and CA3c and became progressively less toward region CA2. Intracellular recordings indicated that in 8.5 mM potassium, large spontaneous excitatory postsynaptic potentials (EPSPs), large burst afterhyperpolarizations, and rhythmic hyperpolarizing-depolarizing waves of membrane potential were invariably present in CA3c neurons. High potassium (8.5 mM) induced a positive shift (+9 mV) in the reversal potential of GABAergic inhibitory postsynaptic potentials (IPSPs) in CA3c neurons without changing input resistance or resting potential. This resulted in a drastic reduction in amplitude of the IPSP. Reduction of IPSP amplitude occurred before the onset of spontaneous bursting and was reversible upon return to normal potassium. A new technique to quantify the relative intensity of interictal-like burst discharges is described. Pentobarbital, diazepam, and GABA uptake inhibitors, which enhance GABA-mediated synaptic inhibition, reduced the intensity of potassium-induced bursts, whereas the GABA antagonist bicuculline increased burst intensity. Diphenylhydantoin and phenobarbital, anticonvulsants that have little effect on GABAergic inhibition, were without effect on spontaneous bursts. Burst frequency was reduced by bicuculline and 4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol but was unaffected by other drugs. Reduction of slice temperature from 35 to 19 degrees C dramatically reduced burst intensity but did not markedly affect burst frequency. We hypothesize that high potassium induces a rise in intracellular chloride concentration, possibly by activating an inward KCl pump or by a passive Donnan effect, which results in a decreased IPSP amplitude. With inhibition suppressed, the large spontaneous EPSPs that appear in high potassium cause individual CA3c neurons to fire. A combination of synaptic and electrical interactions among CA3c cells then synchronizes discharges into interictal spike bursts.