Inspiratory Neurons Research Articles

The augmentation of intrinsic laryngeal muscle activity by air-jet stimulation of the nasal cavity in decerebrate cats

The purpose of this study was to examine the functional roles of nasal afferents in modulating the activity of the intrinsic laryngeal muscles. The electromyographic activities of the intrinsic laryngeal muscles and major respiratory muscles were recorded in cats during nasal air-jet stimulation. The activities of brainstem respiratory neurons were also recorded to determine which neurons transmit nasal afferent signals to the intrinsic laryngeal motoneurons. These axonal projections were identified by antidromic activation evoked by stimulation to the spinal cord at C4 level and the laryngeal nerve. The length of the respiratory cycle was prolonged and the diaphragmatic activity was decreased during air-jet stimulation of the nasal cavity. In contrast, the activities of both the intrinsic laryngeal adductor and abductor muscles were increased. Examination of the laryngeal reflexes revealed increase in the activities of intrinsic laryngeal motoneurons during both respiratory phases. Most of the respiratory neurons recorded decreased their peak firing rate during air-jet stimulation, reflecting decreased diaphragmatic activity; however, the peak firing rate of the bulbospinal expiratory neurons in the portion of the ventral respiratory group caudal to the obex did not decrease during stimulation. These findings demonstrate the nasal air-jet stimulation decreases the activities of major inspiratory muscles in order to avoid inspiration of foreign bodies into the nasal cavity and augments the activities of intrinsic laryngeal muscles, enabling prompt elicitation of the laryngeal airway reflex. Our findings also suggest that the nasal afferents suppress the major inspiratory activities by way of brainstem inspiratory neurons, but that the activities of intrinsic laryngeal muscles are controled through undetermined pathway(s) other than the pathway through respiratory neurons.

AMPA receptor activation and phosphatase inhibition affect neonatal rat respiratory rhythm generation.

1. We investigated the role of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptors and their regulation in affecting respiratory-related neurones in a neonatal rat medullary slice that spontaneously generates respiratory-related rhythm and motor output in the hypoglossal (XII) nerve. 2. Bath application of the AMPA receptor antagonist 1-(4-aminophenyl)-4-methyl-7,8-methylenedioxy-5H-2, 3-benzodiazepine (GYKI) completely blocked XII nerve activity, as well as respiratory-related synaptic drives in neurones within the preBötzinger Complex (preBotC), site of rhythm generation in the slice. 3. Local application of GYKI to the preBötC blocked respiratory rhythm. Local application of AMPA to the preBötC increased rhythm frequency and depolarized respiratory-related neurones. 4. In the presence of tetrodotoxin (TTX), GYKI completely blocked the inward current induced by local application of AMPA, but not that induced by kainate. 5. Local application of okadaic acid, a membrane-permeable inhibitor of phosphatase 1 and 2A, to the preBotC increased the frequency of respiratory motor discharge. 6. Intracellular application of microcystin, a membrane-impermeable inhibitor of phosphatase 1 and 2A, enhanced endogenous inspiratory drive and exogenous AMPA-induced current (in the presence of TTX) in preBotC inspiratory neurones. Both the enhanced inspiratory drive and the increased AMPA-induced current were completely blocked by GYKI. 7. We suggest that AMPA receptor activation and AMPA receptor modulation by phosphorylation are crucial for the rhythm generation within the preBötC.

Synaptic potentials in respiratory neurones during evoked phase switching after NMDA receptor blockade in the cat.

1. Blockade of NMDA receptors by dizocilpine impairs the inspiratory off-switch (IOS) of central origin but not the IOS evoked by stimulation of sensory afferents. To investigate whether this difference was due to the effects of different patterns of synaptic interactions on respiratory neurones, we stimulated electrically the superior laryngeal nerve (SLN) or vagus nerve in decerebrate cats before and after i.v. administration of dizocilpine, whilst recording intracellularly. 2. Phrenic nerve responses to ipsilateral SLN or vagal stimulation were: at mid-inspiration, a transient inhibition often followed by a brief burst of activity; at late inspiration, an IOS; and at mid-expiration, a late burst of activity. 3. In all neurones (n = 16), SLN stimulation at mid-inspiration evoked an early EPSP during phase 1 (latency to the arrest of phrenic nerve activity), followed by an IPSP in inspiratory (I) neurones (n = 8) and by a wave of EPSPs in post-inspiratory (PI) neurones (n = 8) during phase 2 (inhibition of phrenic activity). An EPSP in I neurones and an IPSP in PI neurones occurred during phase 3 (brief phrenic burst) following phase 2. 4. Evoked IOS was associated with a fast (phase 1) activation of PI neurones, whereas during spontaneous IOS, a progressive (30-50 ms) depolarization of PI neurones preceded the arrest of phrenic activity. 5. Phase 3 PSPs were similar to those occurring during the burst of activity seen at the start of spontaneous inspiration. 6. Dizocilpine did not suppress the evoked phrenic inhibition and the late burst of activity. The shapes and timing of the evoked PSPs and the changes in membrane potential in I and PI neurones during the phase transition were not altered. 7. We hypothesize that afferent sensory pathways not requiring NMDA receptors (1) terminate inspiration through a premature activation of PI neurones, and (2) evoke a late burst of phrenic activity which might be the first stage of the inspiratory on-switch.

The hypoxic response of neurones within the in vitro mammalian respiratory network.

1. The transverse brainstem slice preparation containing the pre-Bötzinger complex (PBC) was used in mice to study developmental changes of the response of the in vitro respiratory network to hypoxia. This preparation generates at different postnatal stages (postnatal days (P) 0-22) spontaneous rhythmic activity in hypoglossal (XII) rootlets that occur in synchrony with periodic bursts of neurones in the PBC. 2. In slices from P0-4 mice, hypoxia did not significantly affect the amplitude of rhythmic synaptic drive potentials in four of five inspiratory neurones. Hypoxia reduced, but did not suppress, the amplitude of synaptic drive potentials in only one inspiratory neurone. Spike discharge and phasic 'inspiratory' hyperpolarizations of six expiratory neurones were suppressed during hypoxia revealing a phasic 'inspiratory' depolarization. 3. The coupling between rhythmic activity in PBC neurones and XII bursts occurred under control conditions in preparations from P0-4 mice in a 1:1 manner (n = 11) and from mice older than P5 in a 3:1 manner (n = 9). During hypoxia, PBC and XII activity were linked in a 1:1 manner in all slices. 4. In six of fourteen inspiratory PBC neurones, the amplitude of synaptic drive potentials of slices from mice older than P8 was increased during the period of augmentation, reduced during the period of depression and suppressed during a hypoxic response which we refer to as central apnoea. Augmentation led to a weak-to-moderate membrane depolarization which on average was 4.8 +/- 3.7 mV. This depolarization was followed by a hyperpolarization of 6.2 +/- 4.1 mV only in four inspiratory neurones. In the majority of neurones (n = 9), however, membrane depolarization remained stable and was not followed by hyperpolarization. In expiratory neurones (n = 12) from this age group hypoxia suppressed phasic hyperpolarizations that occurred in synchrony with XII bursts. As similarly seen in inspiratory neurones, membrane potentials were depolarized by 5.1 +/- 4.1 mV during the period of hypoxic augmentation. 5. The hypoxic response of respiratory neurones within the pre-Bötzinger complex resembles the response of neurones that were previously described under in vivo conditions. Thus we conclude that the 'transverse rhythmic slice' is a good model for studying the hypoxic response of the respiratory network under in vitro conditions.

Possible synaptic connections of expiratory neurons in the medulla of newborn rat in vitro.

To elucidate synaptic interactions between expiratory (Exp) and other respiratory neurons in the ventrolateral medulla of brain stem-spinal cord preparations from newborn rats, we analyzed postsynaptic potentials in Exp and a subclass of inspiratory (Insp) neurons using whole-cell recordings. About 72% of the Exp neurons (Exp-p-i) showed Cl--dependent synaptic inhibition during the Insp and pre/post-Insp phases, corresponding to the active phase of the Insp and pre-inspiratory (Pre-I) neurons, respectively. The other 28% neurons (Exp-i) received Cl--dependent inhibition during the Insp phase only. Some Insp neurons showed reversed IPSPs during the active phase of Exp-p-i after Cl-loading. We suggest the existence of inhibitory connections from Pre-I and Insp to Exp and from Exp to Insp neurons. Basic synaptic connections among respiratory neurons similar to those in intact adult mammals may already exist in newborn rats.

Identification and connections of inspiratory premotor neurons in songbirds and budgerigar

Identification and connections of inspiratory premotor neurons in songbirds and budgerigar

Identification and connections of inspiratory premotor neurons in songbirds and budgerigar

Recordings of extracellular unit activity in the ventrolateral medulla and of electromyographic activity in either the M. scalenus, a principal inspiratory muscle, or the abdominal expiratory muscles, were used to identify inspiratory related (IR) neurons. IR neurons extended from levels caudal to the obex through the caudal level of the descending vestibular nucleus. This distribution was found to correspond to that of a subset of cells retrogradely labeled from injections of neuronal tracers into the upper thoracic spinal cord, where motoneurons innervating the M. scalenus were located by retrograde transport. Injections of biotinylated dextran amine at the recording sites resulted in projections to the spinal cord and brainstem. Bulbospinal axons traveled in the lateral funiculus, predominantly contralaterally, and terminated in relation to the dendrites and cell bodies of motoneurons innervating the M. scalenus. Brainstem nuclei receiving projections from injections at IR loci included the retroambigualis, tracheosyringeal motor nucleus, ventrolateral nucleus of the rostral medulla, infraolivaris superior, ventrolateral parabrachial nucleus, and the dorsomedial nucleus of the intercollicular complex. In the finches, there were also bilateral projections to nucleus uvaeformis of the posterior thalamus. The spinal and brainstem projections are similar to those found in pigeon (Reinke and Wild, [1997] J. Comp. Neurol. 379:347-362), and probably mediate the intricate coordination of the vocal (syringeal) and respiratory systems for the control of vocalization. The distribution of IR neurons in birds is similar to that of the rostral ventral respiratory group (rVRG) in mammals.

Effects of NMDA and non-NMDA receptor antagonists on the medullary inspiratory neuronal activity during spontaneous augmented breaths in anesthetized rats

To elucidate whether there is a difference between the effects of iontophoretically applied N-methyl- d-aspartate (NMDA) and non-NMDA receptor antagonists on the activity of inspiratory neurons during spontaneous augmented breaths, extracellular single unit recording of inspiratory neurons (I-augmenting, I-decrementing and I-other) was performed in pentobarbital anesthetized rats. The spontaneous augmented breath was divided into two different phases; the first phase (phase I) resembled a normal inspiration, but the second phase (phase II) consisted of a marked increase in diaphragm electromyogram activity. The mean firing frequency of I-aug type neurons was significantly decreased after 50 nA application of both d-2-amino-5-phosphonopentanoic acid (AP-5) (NMDA receptor antagonist) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) (non-NMDA receptor antagonist). The mean firing frequency of both I-dec and I-other neurons was significantly decreased by both AP-5 and CNQX applications (70 nA). After AP-5 application, relative changes in the discharge rates during inspiratory phases I and II of spontaneous augmented breaths were significantly suppressed in all types of neurons, but CNQX application had no significant effect on the response changes during phase II. In all cell types of neurons, a significant difference between the iontophoretic AP-5 and CNQX applications in the relative mean firing rate was observed. These results suggested that activation of the NMDA receptor-induced neurotransmission can modify the discharge rate of medullary inspiratory neurons, irrespective of the cell types, during the inspiratory phase II of spontaneous augmented breaths, but that non-NMDA receptor blockade may not significantly influence their discharge rate.

Involvement of glycine receptors in coordination between inspiratory and expiratory motor activities in the newborn rat in vitro

Nociceptin/orphanin FQ (N/OFQ) is the endogenous agonist of the N/OFQ peptide receptor, an inhibitory G protein-coupled receptor. N/OFQ acts as a neuromodulator to depress respiratory rhythm in the brainstem. Although the mechanisms of respiratory rhythm generation remain poorly understood, the pre-inspiratory neuron (Pre-I) and the pre-Bötzinger complex (preBötC) inspiratory neuron (Insp) network in the rostral ventrolateral medulla (RVLM) have been proposed to be essential for respiratory rhythm generation. Opioids presumably cause quantal slowing via selective depression of preBötC Insps. However, it is unclear whether N/OFQ depresses respiratory rhythm via the same mechanism. In this study, using in vitro newborn rat en bloc preparations, we examined the slowing pattern of N/OFQ (quantal or non-quantal) and the effects of N/OFQ on the extracellularly recorded discharge of Pre-Is and Insps in the RVLM. N/OFQ caused non-quantal slowing with a synchronous decrease in burst rates of Insps and of C4 discharge whereas the intraburst spike number in Insps remained unchanged. It also caused a significant decrease in burst rates and intraburst spike numbers in Pre-Is, while the 1:1 coupling of Pre-Is bursts to C4 bursts was preserved. When superfusate K+ was elevated from 6.2 to 11.2 mM, Pre-I activity was increasingly uncoupled from C4 bursts. After the application of N/OFQ in a high [K+] superfusate, the 1:1 coupling of Pre-Is to C4 bursts was restored. We conclude that N/OFQ suppresses burst and spike generation of Pre-Is, and that suppression of Pre-Is activity with synchronous coupling to the Insps network contributes to N/OFQ-induced non-quantal slowing.

Synaptic connections to phrenic motoneurons in the decerebrate rat.

Phrenic motoneuron membrane potential trajectories in decerebrate rats exhibit three stages; depolarisation during inspiration, a decreased depolarisation during early expiration and hyperpolarization during late expiration. These trajectories are a result of excitation by ventral-group medullary inspiratory neurons and upper-cervical inspiratory neurons during inspiration and the early part of expiration, and inhibition from Bötzinger-complex expiratory neurons during the late part of expiration.

The site of the antiemetic action of tachykinin NK 1 receptor antagonists may exist in the medullar area adjacent to the semicompact part of the nucleus ambiguus

Nociceptin/orphanin FQ (N/OFQ) is the endogenous agonist of the N/OFQ peptide receptor, an inhibitory G protein-coupled receptor. N/OFQ acts as a neuromodulator to depress respiratory rhythm in the brainstem. Although the mechanisms of respiratory rhythm generation remain poorly understood, the pre-inspiratory neuron (Pre-I) and the pre-Bötzinger complex (preBötC) inspiratory neuron (Insp) network in the rostral ventrolateral medulla (RVLM) have been proposed to be essential for respiratory rhythm generation. Opioids presumably cause quantal slowing via selective depression of preBötC Insps. However, it is unclear whether N/OFQ depresses respiratory rhythm via the same mechanism. In this study, using in vitro newborn rat en bloc preparations, we examined the slowing pattern of N/OFQ (quantal or non-quantal) and the effects of N/OFQ on the extracellularly recorded discharge of Pre-Is and Insps in the RVLM. N/OFQ caused non-quantal slowing with a synchronous decrease in burst rates of Insps and of C4 discharge whereas the intraburst spike number in Insps remained unchanged. It also caused a significant decrease in burst rates and intraburst spike numbers in Pre-Is, while the 1:1 coupling of Pre-Is bursts to C4 bursts was preserved. When superfusate K+ was elevated from 6.2 to 11.2 mM, Pre-I activity was increasingly uncoupled from C4 bursts. After the application of N/OFQ in a high [K+] superfusate, the 1:1 coupling of Pre-Is to C4 bursts was restored. We conclude that N/OFQ suppresses burst and spike generation of Pre-Is, and that suppression of Pre-Is activity with synchronous coupling to the Insps network contributes to N/OFQ-induced non-quantal slowing.

Modulation of respiratory rhythm by 5-HT in the brainstem-spinal cord preparation from newborn rat.

Effects of 5-hydroxytryptamine (5-HT) on inspiration-related nerve activity and membrane potential of respiratory neurons in the ventrolateral medulla were studied in brainstem-spinal cord preparations isolated from newborn rats. Bath application of 5-100 microM 5-HT induced a biphasic response in inspiratory nerve activity: a transient increase in respiratory frequency followed by a decrease in the rate of discharge. The excitatory effect of 5-HT was particularly prominent in preparations with a respiratory rate of less than 3 min-1, whereas the inhibitory effect was more pronounced in preparations with a higher respiratory rate. In pre-inspiratory (Pre-I) and inspiratory (Insp) neurons, 20 microM 5-HT induced a membrane depolarization of up to 10 mV accompanied by a significant decrease in the input resistance. Membrane depolarization by 5-HT was also evident in the presence of tetrodotoxin. In Pre-I neurons, 5-HT caused an increase in the burst rate, which was followed by a decrease in the intraburst firing frequency and burst amplitude, although the burst rate remained high. The burst rate in Insp neurons first increased and subsequently decreased without significant change in the intraburst firing frequency. Simultaneous intra- and extracellular recordings (in the contralateral medulla) of Pre-I/Pre-I neuron or Pre-I/Insp neuron pairs revealed that 5-HT disturbed the correlation between these neuron bursts. Increase in the respiratory rate induced by 20 microM 5-HT was completely blocked by pretreatment (5-15 min) with 5 microM ketanserin or 1 microM methysergide, but not by 10 microM propranolol. None of these antagonists blocked the inhibitory effects of 5-HT. A 5-HT2 agonist, 1-(2, 5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI, 10-100 microM) increased the respiratory rate. Perfusion with a 5-HT1A agonist, 8-hydroxy-dipropylaminotetralin hydrobromide (8-OH-DPAT, 20-100 microM) induced an increase or a decrease in the respiratory rate. A 5-HT2C agonist, 1-(3-chlorophenyl)piperazine (m-CPP, 2-10 microM) induced an initial decrease in the respiratory rate followed by a further long- lasting decrease. Burst activity of Pre-I neurons was suppressed upon administration of 10 microM m-CPP and enhanced with 20 microM DOI. The results suggest that changes in the bursting properties of Pre-I and Insp neurons induced by 5-HT lead to modulation of the respiratory network, thus causing biphasic modulation of the respiratory rhythm. In addition to effects via 5-HT1A receptors, activation of 5-HT2A and 5-HT2C receptor subtypes might be involved in excitatory effects and inhibitory effects of 5-HT respectively.

Axonal projections from the pontine parabrachial-Kölliker-Fuse nuclei to the Bötzinger complex as revealed by antidromic stimulation in cats.

The pontine parabrachial and Kölliker-Fuse nuclear complex (NPB-KF) has been assumed to be the anatomical location of the pneumotaxic center (11). It contains high density of respiratory neurons with various discharge patterns (3, 6). Destruction or electrical stimulation of this area is known to produce profound changes in the respiratory rhythm (5, 24, 25). It is suggested that the NPB-KF exerts its effects by specifically modulating the activity of the medullary inspiratory ‘off-switch’ mechanism, which terminates inspiration and ensures the phase transition from inspiration to expiration (8, 9). However, axonal projections from the NPB-KF to the medullary structures specifically involved in the inspiratory ‘off-switch’ effects have not been well delineated. Previous studies by us demonstrated that the nucleus raphe magnus, a structure involved in the inspiratory ‘off-switch’, received strong axonal projections from the NPB-KF area (1, 21, 22). Another possible neuron group which may transmit the function of the NPB-KF is the Bötzinger complex (Böte). The Böt. c. is known as a group of expiratory neurons in the vicinity of the retrofacial nucleus (14, 19). Most of those neurons have an augmenting firing pattern and widespread inhibitory connections to the medullary inspiratory premotor neurons (10, 12, 18). Recent studies by Smith et al. (20) and us (23) with the retrograde WGA-HRP tracing method revealed that the Böt. c. received strong projections from the lateral NPB and the KF. These studies suggest that these projections are composed of axons from the pontine respiratory and/or non-respiratory neurons which would modulate the activities of expiratory neurons in the Böt. c.KeywordsAxonal ProjectionNucleus ParabrachialisRespiratory NeuronAxonal ArborizationVentral Respiratory GroupThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Entrainment, instability, quasi-periodicity, and chaos in a compound neural oscillator.

We studied the dynamical behavior of a class of compound central pattern generator (CPG) models consisting of a simple neural network oscillator driven by both constant and periodic inputs of varying amplitudes, frequencies, and phases. We focused on a specific oscillator composed of two mutually inhibiting types of neuron (inspiratory and expiratory neurons) that may be considered as a minimal model of the mammalian respiratory rhythm generator. The simulation results demonstrated how a simple CPG model--with a minimum number of neurons and mild nonlinearities--may reproduce a host of complex dynamical behaviors under various periodic inputs. In particular, the network oscillated spontaneously only when both neurons received adequate and proportionate constant excitations. In the presence of a periodic source, the spontaneous rhythm was overridden by an entrained oscillation of varying forms depending on the nature of the source. Stable entrained oscillations were inducible by two types of inputs: (1) anti-phase periodic inputs with alternating agonist-antagonist drives to both neurons and (2) a single periodic drive to only one of the neurons. In-phase inputs, which exert periodic drives of similar magnitude and phase relationships to both neurons, resulted in varying disruptions of the entrained oscillations including magnitude attenuation, harmonic and phase distortions, and quasi-periodic interference. In the absence of significant phasic feedback, chaotic motion developed only when the CPG was driven by multiple periodic inputs. Apneic episodes with repetitive alternation of active (intrinsic oscillation) and inactive (cessation of oscillation) states developed when the network was driven by a moderate periodic input of low frequency. Similar results were demonstrated in other, more complex oscillator models (that is, half-center oscillator and three-phase respiratory network model). These theoretical results may have important implications in elucidating the mechanisms of rhythmogenesis in the mature and developing respiratory CPG as well as other compound CPGs in mammalian and invertebrate nervous systems.

Brainstem neurons projecting to the rostral ventral respiratory group (rVRG) in the medulla oblongata of the rat revealed by co-application of NMDA and biocytin

Groups of neurons in the medulla and pons are essential for the rhythm generation, pattern formation and modulation of respiration. The rostral Ventral Respiratory Group (rVRG) is thought to be a crucial area for rhythm generation. Here we co-applied biocytin and NMDA in the rVRG to label retrogradely brainstem neurons reciprocally connected to a population of inspiratory neurons in the rat rVRG. The procedure excited rVRG neurons in multi-unit recordings and led to a Golgi-like labelling of distant cells presumably excited by efferents from the rVRG. Injection of biocytin without NMDA did not label neurons in distant structures. Several brainstem ipsi- and contralateral structures were found to project to the rVRG, but three major respiratory-related structures, the nucleus of the solitary tract (NTS), the parabrachialis medialis and Kölliker–Fuse nuclei (PB/KF) and the caudal VRG, which are known to project bilaterally to the rVRG, were exclusively labelled ipsilaterally, suggesting an ipsilateral excitation of these structures by the rVRG. The pathways of efferent axons from labelled neurons in the rVRG were traced rostrally towards the pons and caudally to the spinal cord. Terminal axonal arborizations were seen in the same regions where retrogradely filled neurons were found as well as in a few other motor nuclei (the dorsal vagal motor nucleus and XII nucleus). Moreover, in the NTS and the PB/KF, efferent terminal varicosities were seen closely apposed to the soma and proximal dendrites of labelled neurons, suggesting monosynaptic connections between the rVRG and these nuclei.

Synchronization of ventral-group, bulbospinal inspiratory neurons in the decerebrate rat.

We examined the synaptic connections between ventral-group, bulbospinal inspiratory neurons in 27 vagotomized, paralyzed, ventilated, and decerebrated rats using cross-correlation and spike-triggered averaging of intracellular potentials. The neurons were recorded in the medulla about the level of the obex and identified by their inspiratory firing pattern and antidromic activation from the spinal cord at C7. Whole C5 phrenic nerve recordings were made using bipolar electrodes from the central cut ends of the nerve. Most (108/137, 79%) inspiratory neurons discharged only during inspiration but some (29/137, 21%) also discharged during early expiration. Their intracellular membrane potentials displayed a pattern of depolarization during inspiration, repolarization during early expiration, and hyperpolarization during late expiration. Intracellular chloride iontophoresis changed the inspiratory membrane potential trajectories from augmenting to decrementing in 11 of 19 neurons tested (58%), and demonstrated the presence of both early-decrementing and late-augmenting waves of inhibitory postsynaptic potentials during expiration in 11 of 19 neurons tested (58%). Cross-correlation histograms were computed between pairs of extracellularly recorded neurons to detect short time scale synchronizations indicative of synaptic connections (26 ipsilateral; 23 contralateral). While none of the cross-correlation histograms for contralateral pairs showed peaks, most (23, 88%) of those for ipsilateral pairs showed peaks (mean half-amplitude width +/- SD = 1.3 +/- 0.4 ms) at time zero suggestive of common activation. Some of the latter (6, 23%) showed troughs superimposed on the central peaks (mean half-amplitude width +/- SD = 0.9 +/- 0.2 ms) at short latencies (mean latency +/- SD = 1.8 +/- 1.9 ms) suggestive of inhibition; others (8, 31%) had asymmetrical central peaks and two had bilateral peaks suggesting more complex interconnections. Averages of intracellular membrane potentials of inspiratory neurons (n = 24), triggered by action potentials of a nearby extracellularly recorded inspiratory neuron, were computed to detect synchronized postsynaptic potentials. Over half (16, 67%) showed postsynaptic potentials (mean amplitude +/- SD = 201 +/- 176 microV; mean half-amplitude width +/- SD = 2.3 +/- 0.8 ms) confirming the cross-correlation findings of common excitation. We conclude that in decerebrated rats, ventral-group inspiratory neurons projecting to the C7 spinal segment share powerful, ipsilaterally distributed excitatory inputs which enhance their synchronous activity during inspiration. They also receive inhibition during inspiration and early-decrementing and late-augmenting inhibitory inputs during expiration.

Respiratory rhythm generation in mammals: synaptic and membrane properties

Respiratory rhythm generation depends on a complex interaction between synaptic and membrane properties of functionally defined neurons. To gain a better understanding of how inhibitory and excitatory synaptic inputs lead to the generation of the respiratory rhythm we analyzed the depolarization pattern of respiratory neurons that were recorded in the transverse slice preparation of mice (P8–22) and the in vivo adult cat. Using voltage-clamp recordings from respiratory neurons and specific antagonists for inhibitory synaptic transmission we demonstrate under in vitro conditions, that inspiratory ( n=7) and post-inspiratory neurons ( n=13) received concurrent glycinergic and glutamatergic synaptic input during inspiration. A similar conclusion was gained with chloride injections into in vivo respiratory neurons. The inhibitory input was essential not only for generating the characteristic depolarization pattern of respiratory neurons, but also for switching the respiratory rhythm between inspiration and post-inspiration. The generation of the depolarization pattern depends also on intrinsic membrane properties. Negative current injections reveal that excitatory synaptic input was amplified by intrinsic bursting properties in some inspiratory neurons ( n=4) recorded in vitro. Although such properties have not been described under in vivo conditions our findings suggest that with respect to inspiratory, post-inspiratory and late-inspiratory neurons, the principle network organization is similar under both in vitro and in vivo conditions.

Modulation of the synaptic drive to respiratory premotor and motor neurons

The characteristics of GABAergic inhibitory modulation of respiratory bulbospinal neuronal activity and short-term potentiation (STP) of phrenic motoneuronal activity were studied. Extracellular unit recording and picoejection techniques in anesthetized dogs showed that both the spontaneous rhythmic and reflexly induced discharge patterns of inspiratory (I) and expiratory (E) premotor neurons were proportionately amplified by the localized application of picomole amounts of bicuculline (Bic), a competitive GABA A antagonist. Intracellular recording and paired-pulse stimulation techniques in anesthetized rats demonstrated an STP of phrenic motor output that appears to be mediated by NMDA receptors and is associated with facilitation of EPSPs and prolonged depolarization of individual phrenic motoneurons. We speculate that both GABAergic gain modulation of premotor neuronal activity and NMDA-mediated STP of phrenic activity may be neural substrates which are involved with the optimization of respiratory and non-respiratory behaviors, via adaptive and/or differential control of breathing.

Vestibular inputs to bulbar respiratory interneurons in the cat.

Vestibular inputs to medullary respiratory interneurons were studied in decerebrated and artificially ventilated cats. Extracellular recordings were made from 40 neurons located in the area of pre-Bötzinger complex and activated antidromically from the contralateral ventral respiratory group. Neuronal populations analyzed included inspiratory and expiratory neurons with augmenting, constant and decrementing firing patterns, and a late inspiratory neuron. Seventeen neurons responded to ipsilateral and/or contralateral vestibular nerve electrical stimulation. These responses were observed in all seven cell types. Most neuronal reflex responses consisted of inhibition, while a few consisted of either excitation or a combination of both inhibition and excitation. These results indicate that pre-Bötzinger respiratory interneurons, which may be involved in respiratory rhythmogenesis, also participate in vestibulorespiratory responses.

Effect of cardiopulmonary C fibre activation on the firing activity of ventral respiratory group neurones in the rat.

1. Cardiopulmonary C fibre receptor stimulation elicits apnoea and rapid shallow breathing, but the effects on the firing activity of central respiratory neurones are not well understood. This study examined the responses of ventral respiratory group neurones: decrementing expiratory (Edec), augmenting expiratory (Eaug), and inspiratory (I) neurones during cardiopulmonary C fibre receptor-evoked apnoea and rapid shallow breathing. 2. Extracellular neuronal activity, phrenic nerve activity and arterial pressure were recorded in urethane-anaesthetized rats. Cardiopulmonary C fibre receptors were stimulated by right atrial injections of phenylbiguanide. Neurones were tested for antidromic activation from the contra- and ipsilateral ventral respiratory group (VRG), spinal cord and cervical vagus nerve. 3. Edec neurones discharged tonically during cardiopulmonary C fibre-evoked apnoea and rapid shallow breathing, displaying increased burst durations, number of impulses per burst, and mean impulse frequencies. Edec neurones recovered either with the phrenic nerve activity (25 s) or much later (3 min). 4. By contrast, the firing activity of Eaug and most I neurones was decreased, featuring decreased burst durations and number of impulses per burst and increased interburst intervals. Eaug activity recovered in approximately 3 min and inspiratory activity in approximately 1 min. 5. The results indicate that cardiopulmonary C fibre receptor stimulation causes tonic firing of Edec neurones and decreases in Eaug and I neuronal activity coincident with apnoea or rapid shallow breathing.