RESUMO
BACKGROUND: Environmental stress affects the gut with dysmotility being a common consequence. Although a variety of microbes or molecules may prevent the dysmotility, none reverse the dysmotility. METHODS: We have used a 1 hour restraint stress mouse model to test for treatment effects of the neuroactive microbe, L. rhamnosus JB-1™ . Motility of fluid-filled ex vivo gut segments in a perfusion organ bath was recorded by video and migrating motor complexes measured using spatiotemporal maps of diameter changes. KEY RESULTS: Stress reduced jejunal and increased colonic propagating contractile cluster velocities and frequencies, while increasing contraction amplitudes for both. Luminal application of 10E8 cfu/mL JB-1 restored motor complex variables to unstressed levels within minutes of application. L. salivarius or Na.acetate had no treatment effects, while Na.butyrate partially reversed stress effects on colonic frequency and amplitude. Na.propionate reversed the stress effects for jejunum and colon except on jejunal amplitude. CONCLUSIONS & INFERENCES: Our findings demonstrate, for the first time, a potential for certain beneficial microbes as treatment of stress-induced intestinal dysmotility and that the mechanism for restoration of function occurs within the intestine via a rapid drug-like action on the enteric nervous system.
Assuntos
Motilidade Gastrointestinal/fisiologia , Lacticaseibacillus rhamnosus , Probióticos/administração & dosagem , Estresse Psicológico/dietoterapia , Estresse Psicológico/fisiopatologia , Animais , Gastroenteropatias/dietoterapia , Gastroenteropatias/fisiopatologia , Motilidade Gastrointestinal/efeitos dos fármacos , Masculino , Camundongos , Complexo Mioelétrico Migratório/efeitos dos fármacos , Complexo Mioelétrico Migratório/fisiologia , Técnicas de Cultura de Órgãos , Restrição Física/efeitos adversosRESUMO
After a long period of inconclusive observations, the intrinsic primary afferent neurons of the intestine have been identified. The intestine is thus equipped with two groups of afferent neurons, those with cell bodies in cranial and dorsal root ganglia, and these recently identified afferent neurons with cell bodies in the wall of the intestine. The first, tentative, identification of intrinsic primary afferent neurons was by their morphology, which is type II in the terminology of Dogiel. These are multipolar neurons, with some axons that project to other nerve cells in the intestine and other axons that project to the mucosa. Definitive identification came only recently when action potentials were recorded intracellularly from Dogiel type II neurons in response to chemicals applied to the lumenal surface of the intestine and in response to tension in the muscle. These action potentials persisted after all synaptic transmission was blocked, proving the Dogiel type II neurons to be primary afferent neurons. Less direct evidence indicates that intrinsic primary afferent neurons that respond to mechanical stimulation of the mucosal lining are also Dogiel type II neurons. Electrophysiologically, the Dogiel type II neurons are referred to as AH neurons. They exhibit broad action potentials that are followed by early and late afterhyperpolarizing potentials. The intrinsic primary afferent neurons connect with each other at synapses where they transmit via slow excitatory postsynaptic potentials, that last for tens of seconds. Thus the intrinsic primary afferent neurons form self-reinforcing networks. The slow excitatory postsynaptic potentials counteract the late afterhyperpolarizing potentials, thereby increasing the period during which the cells can fire action potentials at high rates. Intrinsic primary afferent neurons transmit to second order neurons (interneurons and motor neurons) via both slow and fast excitatory postsynaptic potentials. Excitation of the intrinsic primary afferent neurons by lumenal chemicals or mechanical stimulation of the mucosa appears to be indirect, via the release of active compounds from endocrine cells in the epithelium. Stretch-induced activation of the intrinsic primary afferent neurons is at least partly dependent on tension generation in smooth muscle, that is itself sensitive to stretch. The intrinsic primary afferent neurons of the intestine are the only vertebrate primary afferent neurons so far identified with cell bodies in a peripheral organ. They are multipolar and receive synapses on their cell bodies, unlike cranial and spinal primary afferent neurons. They communicate with each other via slow excitatory synaptic potentials in self reinforcing networks and with interneurons and motor neurons via both fast and slow EPSPs.
Assuntos
Sistema Nervoso Entérico/citologia , Intestinos/inervação , Neurônios Aferentes/classificação , Animais , Sistema Nervoso Entérico/anatomia & histologiaRESUMO
BACKGROUND: The microbiome is essential for normal myenteric intrinsic primary afferent neuron (IPAN) excitability. These neurons control gut motility and modulate gut-brain signaling by exciting extrinsic afferent fibers innervating the enteric nervous system via an IPAN to extrinsic fiber sensory synapse. We investigated effects of germ-free (GF) status and conventionalization on extrinsic sensory fiber discharge in the mesenteric nerve bundle and IPAN electrophysiology, and compared these findings with those from specific pathogen-free (SPF) mice. As we have previously shown that the IPAN calcium-dependent slow afterhyperpolarization (sAHP) is enhanced in GF mice, we also examined the expression of the calcium-binding protein calbindin in these neurons in these different animal groups. METHODS: IPAN sAHP and mesenteric nerve multiunit discharge were recorded using ex vivo jejunal gut segments from SPF, GF, or conventionalized (CONV) mice. IPANs were excited by adding 5 µM TRAM-34 to the serosal superfusate. We probed for calbindin expression using immunohistochemical techniques. KEY RESULTS: SPF mice had a 21% increase in mesenteric nerve multiunit firing rate and CONV mice a 41% increase when IPANs were excited by TRAM-34. For GF mice, this increase was barely detectable (2%). TRAM-34 changed sAHP area under the curve by -77 for SPF, +3 for GF, or -54% for CONV animals. Calbindin-immunopositive neurons per myenteric ganglion were 36% in SPF, 24% in GF, and 52% in CONV animals. CONCLUSIONS & INFERENCES: The intact microbiome is essential for normal intrinsic and extrinsic nerve function and gut-brain signaling.
Assuntos
Potenciais de Ação/fisiologia , Calbindinas/metabolismo , Microbioma Gastrointestinal/fisiologia , Vida Livre de Germes , Jejuno/inervação , Plexo Mientérico/fisiologia , Neurônios Aferentes/fisiologia , Potenciais de Ação/efeitos dos fármacos , Animais , Sistema Nervoso Entérico/efeitos dos fármacos , Sistema Nervoso Entérico/metabolismo , Sistema Nervoso Entérico/fisiologia , Feminino , Jejuno/metabolismo , Jejuno/microbiologia , Camundongos , Plexo Mientérico/efeitos dos fármacos , Plexo Mientérico/metabolismo , Neurônios Aferentes/efeitos dos fármacos , Pirazóis/farmacologia , Organismos Livres de Patógenos EspecíficosRESUMO
AH neurons are intrinsic sensory neurons of the intestine that exhibit two types of slow synaptic event: slow excitatory postsynaptic potentials which increase their excitability for about 2-4 min, and sustained slow postsynaptic excitation which can persist for several hours, and may be involved in long-term changes in the sensitivity of the intestine to sensory stimuli. The effects of the neurokinin-3 tachykinin receptor antagonist, SR142801, on these two types of synaptic event in AH neurons of the myenteric ganglia of guinea-pig small intestine were compared. Slow excitatory postsynaptic potentials were evoked by stimulation of synaptic inputs at 10-20 Hz for 1s, and sustained slow postsynaptic excitation was evoked by stimulation of inputs at 1Hz for 4 min. SR142801 (1microM) reduced the amplitude of the slow excitatory postsynaptic potential to 26% of control, and also reduced the increase in input resistance and the extent of anode break excitation associated with the slow excitatory postsynaptic potential. In contrast, SR142801 did not reduce the increase in excitability, the increase in input resistance or the depolarisation that occur during the sustained slow postsynaptic excitation. SR142801 did not change the resting membrane potential or the resting input resistance. We conclude that tachykinins, acting through neurokinin-3 receptors, are involved in the generation of the slow excitatory postsynaptic potential, but not in the sustained slow postsynaptic excitation, and that the release of transmitters from synaptic inputs to AH neurons is frequency coded.
Assuntos
Potenciais Pós-Sinápticos Excitadores/fisiologia , Plexo Mientérico/efeitos dos fármacos , Neurônios Aferentes/efeitos dos fármacos , Receptores da Neurocinina-3/antagonistas & inibidores , Receptores da Neurocinina-3/metabolismo , Transmissão Sináptica/efeitos dos fármacos , Taquicininas/metabolismo , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Animais , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Feminino , Cobaias , Intestino Delgado/efeitos dos fármacos , Intestino Delgado/inervação , Intestino Delgado/metabolismo , Masculino , Plexo Mientérico/citologia , Plexo Mientérico/metabolismo , Neurônios Aferentes/metabolismo , Piperidinas/farmacologia , Transmissão Sináptica/fisiologiaRESUMO
Previous experiments have shown that prolonged low frequency stimulation of presynaptic inputs causes excitation of AH neurones that considerably outlasts the period of stimulation in the guinea-pig small intestine. The present experiments compare the responses of S neurones (which are motor neurones and interneurones) with responses of AH neurones (intrinsic primary afferent neurones) to low frequency stimulation of synaptic inputs. Neurones in the myenteric plexus of isolated segments of guinea-pig small intestine were recorded from with intracellular microelectrodes. During their impalement, the neurones were filled with a marker dye and they were later processed to reveal their shapes and immunohistochemical properties. One group of neurones, inhibitory motor neurones to the circular muscle, was depolarised by stimulation of synaptic inputs at 1 Hz for 100 s to 4 min. With 4-min trains of stimuli, peak depolarisation was 21+/-2 mV (mean+/-S.E.M.), which was reached at about 110 s. Depolarisation was accompanied by increased excitability; before stimulation, a test intracellular pulse (500 ms) triggered 3 action potentials, at the peak of excitability this reached 16 action potentials. Depolarisation began to decline immediately at the end of stimulation. This contrasts with responses of AH neurones, in which depolarisation persisted after the end of the stimulus (peak depolarisation at 300 s). The excitation and depolarisation of inhibitory motor neurones was blocked by the neurokinin 1 tachykinin receptor antagonist, SR140333 (100 nM), but excitation of AH neurones was not affected. Small or no responses to 1 Hz stimulation were recorded from descending filamentous interneurones, longitudinal muscle motor neurones and excitatory circular muscle motor neurones. In conclusion, this study indicates that sustained slow postsynaptic excitation only occurs in AH neurones, and that one type of S neurones, inhibitory motor neurones to the circular muscle, responds substantially, but not beyond the period of stimulation, to activation of synaptic inputs at 1 Hz. This slow excitatory postsynaptic potential evoked by low frequency stimulation is mediated by tachykinins.
Assuntos
Plexo Mientérico/fisiologia , Inibição Neural/fisiologia , Vias Neurais/fisiologia , Neurônios/fisiologia , Sinapses/fisiologia , Transmissão Sináptica/fisiologia , Taquicininas/metabolismo , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Animais , Axônios/efeitos dos fármacos , Axônios/fisiologia , Axônios/ultraestrutura , Tamanho Celular/efeitos dos fármacos , Tamanho Celular/fisiologia , Estimulação Elétrica , Feminino , Bloqueadores Ganglionares/farmacologia , Cobaias , Hexametônio/farmacologia , Interneurônios/citologia , Interneurônios/efeitos dos fármacos , Interneurônios/fisiologia , Masculino , Neurônios Motores/citologia , Neurônios Motores/efeitos dos fármacos , Neurônios Motores/fisiologia , Plexo Mientérico/citologia , Plexo Mientérico/efeitos dos fármacos , Condução Nervosa/efeitos dos fármacos , Condução Nervosa/fisiologia , Inibição Neural/efeitos dos fármacos , Vias Neurais/citologia , Vias Neurais/efeitos dos fármacos , Neurônios/citologia , Neurônios/efeitos dos fármacos , Receptores de Taquicininas/antagonistas & inibidores , Receptores de Taquicininas/metabolismo , Sinapses/efeitos dos fármacos , Sinapses/ultraestrutura , Transmissão Sináptica/efeitos dos fármacos , Taquicininas/antagonistas & inibidores , Tetrodotoxina/farmacologiaRESUMO
It is commonly believed that the cell bodies of mammalian sensory neurons are contained within spinal and cranial sensory ganglia associated with the central nervous system or within the central nervous system itself. However, strong circumstantial evidence implies that some sensory neurons are contained entirely within the gastrointestinal tract. We have investigated this possibility by using intracellular methods to record the responses of myenteric neurons in the guinea-pig small intestine to physiological stimuli applied to the neighbouring mucosa. The results show that the myenteric plexus contains a population of chemosensitive sensory neurons and that these neurons correspond to neurons with AH electrophysiological properties and Dogiel type II morphology. This is the first direct evidence that some sensory neurons are contained entirely within the peripheral nervous system.
Assuntos
Intestinos/inervação , Neurônios Aferentes/fisiologia , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Animais , Eletrofisiologia , Cobaias , Mucosa Intestinal/inervação , Plexo Mientérico/citologia , Plexo Mientérico/fisiologia , Sinapses/fisiologiaRESUMO
Simultaneous intracellular electrical recordings were made from pairs of neurons separated circumferentially by 100-200 microns of the myenteric plexus of the guinea-pig ileum in vitro. The recording electrodes were filled with the dye neurobiotin which was injected into impaled nerve cells, and later revealed histochemically. Intracellular current pulses were used to evoke action potentials via the recording electrode in one type of myenteric neuron, in most cases an AH neuron, while a second electrode was used to record from a simultaneously impaled S neuron or AH neuron. AH neurons are thought to be primary sensory neurons, whereas S neurons are interneurons and motor neurons. Ninety pairs of neurons were adequately tested for interaction. From these, 17 S neurons and three AH neurons that responded to AH neuron stimulation were detected. In each case, the response was a slow depolarization that was seen only in response to a train of stimuli at 10 Hz. The slow depolarizations were enhanced by passing depolarizing current and diminished by hyperpolarization. Responses were also diminished by lowering external Ca.2+ and elevating Mg2+. In all cases in which intracellular recording indicated communication between neurons, morphological evidence of connection was seen. In no case was there communication without connection, but in four instances, morphological connections appeared to exist, although no physiological evidence of communication was obtained.
Assuntos
Potenciais de Ação , Plexo Mientérico/fisiologia , Neurônios Aferentes/fisiologia , Potenciais de Ação/efeitos dos fármacos , Animais , Axônios/ultraestrutura , Cálcio/farmacologia , Comunicação Celular , Tamanho Celular , Estimulação Elétrica , Cobaias , Íleo/inervação , Interneurônios/fisiologia , Interneurônios/ultraestrutura , Magnésio/farmacologia , Neurônios Motores/fisiologia , Neurônios Motores/ultraestrutura , Plexo Mientérico/citologia , Neurônios Aferentes/efeitos dos fármacos , Neurônios Aferentes/ultraestrutura , Sinapses/ultraestruturaRESUMO
Intracellular recording, dye filling and immunohistochemistry were used to investigate neurons of the proximal duodenum of the guinea-pig. Recordings were made from neurons of the myenteric plexus in the presence of nicardipine to quell muscle contractions, using microelectrodes that contained the marker substance Neurobiotin. Preparations were subsequently processed histochemically to reveal nerve cell shapes and immunoreactivity for calbindin, calretinin or nitric oxide synthase. Neurons were distinguished by their shapes and axonal projections as Dogiel type II, Dogiel type I, filamentous descending interneurons and small filamentous neurons. Dogiel type II cells had large cell bodies and multiple axon processes. They each had a broad action potential (mean half-width, 2.9 ms) and a prominent inflection (hump) on the falling phase of the action potential. The majority (70%) of Dogiel type II cells were AH neurons, defined by their having a prolonged hyperpolarizing potential that followed a soma action potential and lasted more than 2 s. Fast excitatory postsynaptic potentials were not recorded from Dogiel type II neurons. Two thirds of Dogiel type II neurons fired phasically in response to intracellularly injected 500 ms depolarizing current pulses and one-third fired tonically. Calbindin immunoreactivity occurred in 70% of Dogiel type II neurons. Dogiel type I neurons had lamellar dendrites and a single axon. They had brief action potentials (mean half-width, 1.7 ms) with no, or a slight hump. They responded to fibre tract stimulation with fast excitatory postsynaptic potentials. Only 2/21 exhibited a prolonged hyperpolarization following action potentials. The majority of Dogiel type I neurons thus belong to the S neuron category. Nine Dogiel type I neurons fired phasically in response to 500 ms depolarizing current pulses, while 12 fired tonically. Filamentous descending interneurons had long, branching filamentous dendrites and a single anally-projecting axon which gave rise to varicose branches in myenteric ganglia. Action potential characteristics of filamentous interneurons ranged between those of Dogiel type II and type I neurons. Small neurons. Small neurons with short filamentous, or few simple dendrites were also characterized. They had single axons, which could be traced either locally to the circular muscle, or to the longitudinal muscle. None of 12 filamentous interneurons or of 10 small filamentous neurons exhibited a prolonged post-spike hyperpolarization, whereas fast excitatory postsynaptic potentials were recorded from a majority. It is concluded that the morphological types of neuron that are encountered in the ileum also occur in the duodenum, but the electrophysiological characteristics of the neurons are more variable for each morphological class. Thus, it is not always possible to predict the morphology of myenteric neurons in the duodenum from their electrophysiological properties. Part of the electrophysiological variability appears to be due to duodenal neurons being more excitable than ileal neurons.
Assuntos
Duodeno/inervação , Plexo Mientérico/citologia , Plexo Mientérico/fisiologia , Neurônios/fisiologia , Potenciais de Ação/fisiologia , Animais , Biotina/análogos & derivados , Biotina/metabolismo , Calbindinas , Dendritos/fisiologia , Duodeno/anatomia & histologia , Duodeno/fisiologia , Estimulação Elétrica , Eletrofisiologia , Cobaias , Imuno-Histoquímica , Interneurônios/fisiologia , Potenciais da Membrana/fisiologia , Proteínas do Tecido Nervoso/metabolismo , Neurônios/ultraestrutura , Técnicas de Patch-Clamp , Proteína G de Ligação ao Cálcio S100/metabolismoRESUMO
The effects of centrifugal afferents on single unit discharge in the main olfactory bulb were studied in anaesthetized rats. Recording with extracellular micropipettes revealed spontaneous firing in all bulb layers. Units were located to different laminae using evoked field-potential profiles and histological verification. Output neurons were identified by antidromic response to stimulation of the lateral olfactory tract. Single- or brief multiple-pulse stimulation in the nucleus of the horizontal limb of the diagonal band, but not in adjacent regions, facilitated 17 out of 27 mitral cells with no effect on 10, but inhibited 21 out of 33 granule cell layer units with no effect on 12. Of 13 presumed tufted cells, six were facilitated and the rest unaffected. In contrast, stimulation of olfactory cortex inhibited mitral cells and facilitated most granule layer cells. The results are consistent with an inhibition of tonic granule cell discharge by the horizontal diagonal band nucleus, with resultant disinhibition of mitral cells via the dendrodendritic synapses of granule cells on mitral cell secondary dendrites.
Assuntos
Neurônios/fisiologia , Bulbo Olfatório/fisiologia , Animais , Estimulação Elétrica , Potenciais Evocados , Masculino , Bulbo Olfatório/citologia , Ratos , Ratos EndogâmicosRESUMO
The aim of this study was to identify the receptor type(s) by which 5-hydroxytryptamine applied to the intestinal mucosa excites the terminals of myenteric AH neurons. The AH neurons have been identified as the intrinsic primary afferent (sensory) neurons in guinea-pig small intestine and 5-hydroxytryptamine has been identified as a possible intermediate in the sensory transduction process. Intracellular recordings were taken from AH neurons located within 1mm of intact mucosa to which 5-hydroxytryptamine was applied. Trains of action potentials and/or slow depolarizing responses were recorded in AH neurons in response to mucosal application of 5-hydroxytryptamine (10 or 20microM) or the 5-hydroxytryptamine-3 receptor agonist, 2-methyl-5-hydroxytryptamine (1 or 3mM), and to electrical stimulation of the mucosa. The 5-hydroxytryptamine-2 receptor agonist, alpha-methyl-5-hydroxytryptamine (100microM), and the 5-hydroxytryptamine-1,2,4 receptor agonist, 5-methoxytryptamine (10microM), did not elicit such responses. The 5-hydroxytryptamine-3 receptor-selective antagonist, granisetron (typically 1microM), and the 5-hydroxytryptamine-3,4 receptor antagonist, tropisetron (typically 1microM), each reduced or abolished the responses to 5-hydroxytryptamine, while the selective 5-hydroxytryptamine-4 receptor antagonist, SB 204070 (1microM), did not. It is concluded that application of 5-hydroxytryptamine to the mucosa activates a 5-hydroxytryptamine-3 receptor that triggers action potential generation in the mucosal nerve terminals of myenteric AH neurons.
Assuntos
Íleo/inervação , Plexo Mientérico/efeitos dos fármacos , Neurônios Aferentes/efeitos dos fármacos , Terminações Pré-Sinápticas/efeitos dos fármacos , Receptores de Serotonina/efeitos dos fármacos , Serotonina/análogos & derivados , Serotonina/farmacologia , 5-Metoxitriptamina/metabolismo , 5-Metoxitriptamina/farmacologia , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Animais , Dioxanos/farmacologia , Granisetron/farmacologia , Cobaias , Íleo/efeitos dos fármacos , Íleo/fisiologia , Indóis/farmacologia , Plexo Mientérico/citologia , Plexo Mientérico/metabolismo , Neurônios Aferentes/citologia , Neurônios Aferentes/metabolismo , Piperidinas/farmacologia , Terminações Pré-Sinápticas/metabolismo , Terminações Pré-Sinápticas/ultraestrutura , Receptores de Serotonina/metabolismo , Receptores 5-HT3 de Serotonina , Serotonina/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia , TropizetronaRESUMO
The effects of centrifugal afferents on membrane potentials of identified granule cell layer using evoked field potential profiles, and trans-synaptic activation via antidromic stimulation of output cell axon collaterals. Intracellular recordings maintained for 4-30 min showed complex spontaneous spike discharges and allowed characterization of the cell's input resistance, and on some occasions its morphology following intracellular injection of Lucifer Yellow. Stimulation in the nucleus of the horizontal limb of the diagonal band, but not surrounding regions, produced hyperpolarizing responses in 13 of 27 cells in the granule cell layer; four of these were morphologically identified as granule cells of two types, in five the responses had reversal potentials more negative than the resting potential, and six were identified as granule cells by monosynaptic activation from output axon collaterals. A different set of three cells in the granule cell layer responded with depolarization. The results are consistent with the inhibition of tonic activity of granule cells by the nucleus of the horizontal limb of the diagonal band, leading to disinhibition of mitral and tufted cells via dendrodendritic synapses of granule cells on mitral/tufted cell secondary dendrites.
Assuntos
Bulbo Olfatório/fisiologia , Potenciais de Ação , Animais , Axônios/fisiologia , Estimulação Elétrica , Potenciais Evocados , Complexo de Golgi/ultraestrutura , Masculino , Potenciais da Membrana , Bulbo Olfatório/citologia , Ratos , Ratos Endogâmicos , Coloração e Rotulagem , Sinapses/fisiologiaRESUMO
Intracellular microelectrodes were used to examine the active and passive membrane properties of neurons in the myenteric plexus of the guinea-pig small intestine. Neurons of two types were examined: S neurons, which have prominent fast excitatory postsynaptic potentials and in which action potentials are not followed by long-lasting afterhyperpolarizations, and AH neurons, which have long-lasting afterhyperpolarizations following soma action potentials. In preparations in which the myenteric ganglia and longitudinal muscle, but no mucosa, were present, most S neurons (59/64) responded to intracellular depolarizing current with brief bursts of action potentials. Regardless of the strength of a depolarizing current of 500-ms duration, these neurons never fired action potentials beyond the first 250 ms. S neurons in this state were called rapidly accommodating. In contrast, within 600 microm circumferential to the intact mucosa, 26/58 S neurons fired action potentials for most or all of the period of a 500-ms insightful depolarizing pulse. S neurons in this state were called slowly accommodating. Depolarization of S neurons in the rapidly accommodating state caused a rapidly developing reduction in membrane resistance (outward rectification; onset about 7 ms). This rectification was absent from S neurons in the slowly accommodating state. Tetraethylammonium blocked the early rectification and the changed neuronal state from rapidly accommodating to slowly accommodating. Application of tetrodotoxin to neurons in the slowly accommodating state revealed the early rectification, indicating that its absence from these neurons before tetrodotoxin was applied had been due to ongoing activity in axons providing synaptic input to the neurons. After the mucosa was disconnected from the other layers and laid back in its original position, all S neurons close to the mucosa were in the rapidly accommodating state (17/17). Slow excitatory postsynaptic potentials, evoked by electrical stimulation of nerve tracts, converted 17 of 43 S neurons from rapidly accommodating to slowly accommodating and eliminated the early outward rectification in these neurons. These results indicate that the action potential firing properties of S neurons can be changed by external influences, including the activity of synaptic inputs that release a slowly acting transmitter. Spontaneous antidromic action potentials were recorded in 8/62 AH neurons within 600 microm circumferential to the intact mucosa. It is concluded that, when the mucosa is intact, a background firing of sensory neurons occurs which leads to a state change in many S neurons innervated by the active sensory neurons. We conclude that this state change is caused by the block of a voltage-sensitive outward rectification.
Assuntos
Mucosa Intestinal/inervação , Plexo Mientérico/fisiologia , Neurônios/fisiologia , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Animais , Estimulação Elétrica , Eletrofisiologia , Potenciais Evocados/efeitos dos fármacos , Potenciais Evocados/fisiologia , Cobaias , Íleo/inervação , Íleo/fisiologia , Técnicas In Vitro , Potenciais da Membrana/efeitos dos fármacos , Potenciais da Membrana/fisiologia , Plexo Mientérico/citologia , Sinapses/efeitos dos fármacos , Sinapses/fisiologia , Compostos de Tetraetilamônio/farmacologia , Tetrodotoxina/farmacologiaRESUMO
Intracellular microelectrodes were used to record the effects of extended periods (1-30 min) of synaptic activation on AH neurons in the myenteric ganglia of the guinea-pig ileum. Low-frequency (1 Hz) stimulation gave rise to a slowly developing, sustained increase in excitability of the neurons associated with depolarization and increased input resistance. The increased excitability lasted for up to 3.5 h following the stimulus period. Successive stimulus trains (1-4 min) elicited successively greater increases in excitability. The neurons went through stages of excitation. Before stimulation, 500-ms depolarizing pulses evoked up to three action potentials (phasic response) and anode break action potentials were not observed. As excitability increased, more action potentials were evoked by depolarization (the responses became tonic), anode break action potentials were observed, prolonged after hyperpolarizing potentials that follow multiple action potentials were diminished and, with substantial depolarization of the neurons, invasion by antidromic action potentials was suppressed. It is concluded that a state of elevated excitability is induced in myenteric AH neurons by synaptic activation at low frequency and that changes in excitability can outlast stimulation by several hours.
Assuntos
Potenciais de Ação/fisiologia , Neurônios/fisiologia , Sinapses/fisiologia , Animais , Estimulação Elétrica , Cobaias , Íleo/fisiologia , Tempo de Reação/fisiologiaRESUMO
Nerve circuits within the proximal duodenum were investigated using a combination of immunohistochemistry for individual neuron markers and lesion of intrinsic nerve pathways to determine axon projections. Cell shapes and axonal projections were also studied in cells that had been injected with a marker substance. Several major neuron populations were identified. Calbindin immunoreactivity occurred in a population of myenteric nerve cells with Dogiel type II morphology. These had axons that projected to other myenteric ganglia, to the circular muscle and to the mucosa. All were immunoreactive for the synthesizing enzyme for acetylcholine, choline acetyltransferase, and some were also immunoreactive for calretinin. Myenteric neurons with nitric oxide synthase immunoreactivity projected anally to the circular muscle. These were also immunoreactive for vasoactive intestinal peptide, and proportions of them had enkephalin and/or neuropeptide Y immunoreactivity. It is suggested that they are inhibitory motor neurons to the circular muscle. A very few (about 2%) of nitric oxide synthase-immunoreactive neurons had choline acetyltransferase immunoreactivity. Tachykinin (substance P)-immunoreactive nerve cells were numerous in the myenteric plexus. Some of these projected orally to the circular muscle and are concluded to be excitatory motor neurons. Others projected to the tertiary plexus which innervates the longitudinal muscle and others provided terminals in the myenteric plexus. Two groups of descending interneurons were identified, one with somatostatin immunoreactivity and one with vasoactive intestinal peptide immunoreactivity. The two most common nerve cells in submucous ganglia were neuropeptide Y- and vasoactive intestinal peptide-immunoreactive nerve cells. Both provided innervation of the mucosa. There was also a population of calretinin-immunoreactive submucous neurons that innervated the mucosal glands, but not the villi. Comparison with the ileum reveals similarities in the chemistries and projections of neurons. Differences include the almost complete absence of nitric oxide synthase immunoreactivity from vasoactive intestinal peptide-immunoreactive interneurons in the duodenum, the projection of calbindin-immunoreactive Dogiel type II neurons to the circular muscle and the absence of tachykinin-immunoreactivity from these neurons.
Assuntos
Duodeno/inervação , Mucosa Intestinal/inervação , Plexo Mientérico/citologia , Plexo Mientérico/fisiologia , Neurônios/citologia , Neuropeptídeos/análise , Animais , Axônios/fisiologia , Axônios/ultraestrutura , Calbindina 2 , Calbindinas , Colina O-Acetiltransferase/análise , Cobaias , Imuno-Histoquímica/métodos , Proteínas do Tecido Nervoso/análise , Neurônios/fisiologia , Óxido Nítrico Sintase/análise , Óxido Nítrico Sintase Tipo I , Proteína G de Ligação ao Cálcio S100/análiseRESUMO
We have developed methods to examine the neurochemistry of synaptic inputs to individual myenteric neurons labeled by dye injection through intracellular recording electrodes. Myenteric neurons of the guinea-pig ileum were filled with Neurobiotin, fixed, washed in 50% ethanol, exposed to sodium cyanoborohydride, incubated in avidin-biotin-horseradish peroxidase and incubated in antisera to calretinin or calbindin. The Neurobiotin-filled cells were revealed using the diaminobenzidine (DAB) reaction. The tissue was examined at the light microscope level to determine the morphology and projections of the Neurobiotin-filled neurons, and then incubated in 1 nm gold-labeled secondary antibodies. Following osmication, the gold probes were silver-intensified and the tissue embedded flat in resin. The tissue was re-examined at the light microscope level. Neurons containing a DAB reaction product could be distinguished from neurons containing a silver-intensified gold reaction product using oblique or epipolarized illumination. Ultrathin sections were taken through the injected neurons and examined. At the ultrastructural level, Neurobiotin-filled cell bodies and their processes (labeled with DAB) were easily distinguished from the structures labeled by silver-intensified gold. Gold-labeled terminals of enteric interneurons made synapses and close contacts with Neurobiotin-filled nerve cell bodies and their processes. This technique is valuable for the neurochemical identification of synaptic inputs to morphologically and/or functionally characterized myenteric neurons and could be easily applied to other preparations, such as brain slices.
Assuntos
Biotina/análogos & derivados , Ouro , Plexo Mientérico/metabolismo , Neurônios/metabolismo , Prata , Inclusão do Tecido , Animais , Biotina/administração & dosagem , Biotina/metabolismo , Eletrofisiologia , Feminino , Cobaias , Imuno-Histoquímica , Injeções , Masculino , Plexo Mientérico/citologia , Plexo Mientérico/ultraestrutura , Sinapses/ultraestrutura , p-DimetilaminoazobenzenoRESUMO
The patterns of innervation of the mucosa by axons of individual primary afferent neurones with cell bodies in the myenteric plexus were studied by mapping sites from which electrical stimulation of the mucosa elicited action potentials (APs) in their cell bodies. Segments of guinea-pig ileum were dissected to reveal the myenteric plexus over half of the intestinal circumference, leaving the mucosa intact over the other half. Intracellular recordings were taken from myenteric neurones located within 1 mm of the intact mucosa. Focal electrical stimuli were applied to the mucosa at multiple locations separated by about 1 mm. Neurones that responded had round or oval cell bodies with several long processes (Dogiel type II) and APs that had an inflection on the falling phase (AH-neurones). Responses consisted of single APs or bursts of APs. Maps of the mucosal projections of 30 neurones were generated. The maximum distances from which individual neurones responded were 7 mm circumferential and 2 mm oral or anal to the cell body with a higher proportion of responses from the oral regions. The areas of intact mucosa calculated to be innervated ranged from 1 mm2 up to approximately 15 mm2 (mean 3.9 mm2; median 2.5 mm2). It is estimated that the areas innervated would be two to three times larger under conditions where part of the mucosa is not removed. Some neurones also responded to a chemical or a mechanical stimulus applied to the mucosa within the electrically mapped area. It is concluded that intrinsic primary afferent neurones have overlapping receptive fields with 230-350 neurones innervating the same region of mucosa.
Assuntos
Mucosa Intestinal/inervação , Intestino Delgado/inervação , Neurônios Aferentes/fisiologia , Transmissão Sináptica/fisiologia , Animais , Estimulação Elétrica , Eletrofisiologia , Feminino , Cobaias , Masculino , Neurônios Aferentes/citologia , Estimulação Física , Estimulação QuímicaRESUMO
We present intracellular data which demonstrates a unique facilitatory centrifugal influence on the output cells of the olfactory bulb; the source being the lateral component of the nucleus of the horizontal limb of the diagonal band (HDB), part of the basal forebrain magnocellular complex. Damage to this facilitatory HDB influence may explain the loss of olfactory sensitivity seen early in Alzheimer's disease in which pathological changes occur in the basal forebrain.
Assuntos
Neurônios/fisiologia , Bulbo Olfatório/fisiologia , Prosencéfalo/fisiologia , Potenciais de Ação , Animais , Estimulação Elétrica , Potenciais Evocados , Lobo Frontal/fisiologia , Masculino , Ratos , Ratos EndogâmicosRESUMO
Extracellular and intracellular recordings were made from antidromically identified spinocervical tract (SCT) cells in the medial part of the lumbosacral dorsal horn in anesthetized cats. The low threshold mechanoreceptive fields (RFs) of 18 cells were mapped during extracellular recording, and for 15 the RF included an area of glabrous skin. Intracellular recordings were made from 6 of these during electrical microstimulation of glabrous skin and all showed excitatory postsynaptic potentials at latencies consistent with conduction over group II primary afferent fibers. The distance from the medial border of the dorsal horn of the recording loci of 12 cells, 4 of which were injected with horseradish peroxidase, was measured. It is concluded that some medially situated SCT cells receive excitatory input from low threshold group II primary afferent fibers from glabrous skin.
Assuntos
Pé/inervação , Pele/inervação , Medula Espinal/fisiologia , Tato/fisiologia , Animais , Gatos , Potenciais Evocados , Membro Posterior , Medula Espinal/citologiaRESUMO
A theoretical method is described for estimating the distance between a spike recording-site, possibly axonal, and the corresponding cell body of unknown location. The method requires that an orthodromic spike be recorded following an antidromic spike, with estimation of a collision interval analogous to that used for establishing antidromicity. To calculate the distance between recording-site and cell body, values are needed for the collision interval between antidromic and succeeding orthodromic spikes, the refractory period of the spike, and the antidromic conduction speed. Problems may arise in determining the last value. The method is illustrated with antidromic spikes recorded in the medial thalamus of the cat upon stimulating the caudate nucleus.
Assuntos
Axônios/fisiologia , Mapeamento Encefálico/métodos , Condução Nervosa , Neurônios/fisiologia , Animais , Gatos , Núcleo Caudado/fisiologia , Estimulação Elétrica , Vias Neurais/fisiologia , Tálamo/fisiologiaRESUMO
Extracellular recordings from the globus pallidus of awake, unrestrained rats showed a distinctive bursting activity during grooming behaviour and in periods of stereotyped jaw movements induced by amphetamine (3 or 5 mg/kg IP) or apomorphine (2 mg/kg SC). During stereotyped licking, there was one burst for each outward movement of the tongue. The bursts were shown to consist of several separate unit spikes firing so as to produce a fusiform envelope of amplitudes, suggesting an ordered recruitment of pallidal neurons related to licking.