Respiratory Central Pattern Generator Research Articles

Analysis of excitatory and inhibitory interactions at high temporal resolution in core circuits of the respiratory CPG

The pre-Botzinger complex (pre-BotC) is the essential core component of the brainstem respiratory central pattern generator (rCPG) in mammals. Phasic excitatory and inhibitory synaptic inputs during the respiratory cycle are thought to dynamically shape membrane potential trajectories and spiking patterns of pre-BotC neurons. These synaptic inputs reflect the functionally interacting neuron populations involved in rhythm and inspiratory-expiratory pattern generation. The dynamic patterns of these synaptic inputs have not been systematically studied and characterized experimentally. Accurate temporal reconstruction of the synaptic input patterns is important for testing current models of the rCPG that incorporate specific excitatory and inhibitory circuit mechanisms for rhythm and pattern generation. A recent computational model of the rCPG generating a normal “three-phase” respiratory pattern [1] specified patterns of synaptic excitation and inhibition in the preBotC during the respiratory cycle required to produce the behaviorally distinct phases of inspiration (I) and two phases of expiration (E1, or post-inspiration, and E2). This model has not been rigorously tested by experimental data on dynamic changes in phasic postsynaptic excitatory and inhibitory conductances in different populations of pre-BotC inspiratory and expiratory neurons. We developed analytical techniques that allow quantitative reconstruction of the patterns of synaptic conductances at high temporal resolution from intracellular recordings of membrane potential trajectories. Our approach extends previously proposed methods for extracting dynamic patterns of synaptic input conductances [2,3] and provides nearly continuous readouts of inhibitory and excitatory synaptic conductances in electrophysiologically different cell types throughout the respiratory cycle. The membrane potential trajectories of pre-BotC respiratory neurons were obtained by sharp microelectrode current-clamp recording within in situ perfused brainstem-spinal cord preparations of mature rats, which generate a three-phase respiratory pattern and provide mechanically stable conditions for intracellular recordings in functionally intact brainstem circuits. We distinguished different types of inspiratory and expiratory pre-BotC neurons and analyzed the dynamical changes of excitatory (Ge )a nd inhibitory (Gi) conductances. Inspiratory neurons exhibited strong excitatory inputs (large dynamic increases in Ge) during their spiking phase followed by a wave(s) of inhibitory inputs during the expiratory period with large increases in Gi and temporal patterns consistent with two populations of inhibitory neurons providing inputs that coordinate inspiratory to expiratory and expiratory to inspiratory phase transitions. Expiratory neurons exhibited strong inhibition during the inspiratory phase, consistent with the rhythmic alternation of inspiration and expiration, and changes in Ge indicating reciprocal interactions between two major populations of inhibitory neurons coordinating the generation of two (E1 and E2) phases of expiration. The reconstructed patterns of Ge and Gi are generally consistent with previously proposed circuit architecture [1] and also suggest its extension. The techniques that we have developed for high temporal resolution of postsynaptic conductance changes are likely to have broad application for analysis of synaptic interactions in circuit components whenever intracellular recordings of membrane potentials can be

The effects of vestibular stimulation rate and magnitude of acceleration on central pattern generation for chest wall kinematics in preterm infants

To examine the role of vestibular inputs on respiratory and oromotor systems in healthy preterm infants. A total of 27 preterm infants were quasi-randomly assigned to either the VestibuGlide treatment or control groups. VestibuGlide infants were held in a developmentally supportive position, given a pacifier and received a series of vestibular stimuli, counterbalanced across rate and acceleration conditions, 15 min 3 times per day for 10 days. The control infants were also held in a developmentally supportive position, given a pacifier for 15 min 3 times per day for 10 days but did not receive the VestibuGlide stimulation. A multi-level regression model revealed that treatment infants increased their respiratory rate in response to vestibular stimulus, and that the highest level of vestibular acceleration delivered to the infants (0.51 ms(-2)) resulted in a significant increase in breaths per minute. Vestibular stimulation delivered to preterm infants before scheduled feeds effectively modulates respiratory rate and resets the respiratory central pattern generator.

State-dependent control of the respiratory pattern and coupled oscillators

The respiratory rhythm and motor pattern are generated by a spatially organized brain stem respiratory network with a rhythmogenic core comprising interacting neural populations within the pre-Botzinger (pre-BotC) and Botzinger (BotC) complexes controlled by drives and inputs from other brain stem compartments. Our previous large-scale computational model [3, 4] reproduced the behavior of this network under multiple natural and experimental conditions, but did not consider neural oscillations that were proposed to emerge within the retrotrapezoid nucleus / parafacial respiratory group (RTN/pFRG) and drive pre-inspiratory (or late-expiratory, late-E) oscillations in the abdominal motor output [2]. These late-E oscillations usually emerge with increasing metabolic demands (e.g., during hypercapnia or hypoxia) and couple with the respiratory oscillations generated by the BotC-pre-BotC core of the respiratory central pattern generator [1, 2]. In our experiments, performed in the arterially perfused in situ rat preparation, the respiratory motor activity was recorded simultaneously from abdominal, phrenic, hypoglossal and central vagus nerves. Under normal conditions (eupnea), the abdominal motor outflow showed only a low-amplitude post-inspiratory (post-I) activity. An expression of the abdominal late-E bursts, preceding and coupled to phrenic discharges, occurred during hypercapnia (7–10% CO2) and could be abolished by pharmacological inactivation of the RTN/pFRG region and by administration of riluzole, a persistent sodium current (I NaP ) blocker, hence confirming that the late-E oscillations originate in RTN/pFRG and involve intrinsic, I NaP -dependent mechanisms. Using these data we extended our computational model by incorporating in the RTN/pFRG compartment an additional late-E population (with neurons containing I NaP ) serving as a source of abdominal late-E activity. The proposed interactions between the BotC’s and pre-BotC’s populations and the late-E population of RTN/pFRG allowed the model to reproduce several experimentally observed behaviors, including quantal acceleration of abdominal late-E oscillations with progressive hypercapnia and quantal slowing of phrenic activity with progressive suppression of pre-BotC excitability (simulating the suppressing effects of opioids), as well as to predict a release of late-E oscillations by disinhibition of RTN/pFRG under normal conditions. The latter prediction has been confirmed in our experimentally studies. Our experimental and modeling studies suggest that under normal metabolic conditions the RTN/pFRG oscillator is inhibited by both the post-I population of BotC during inspiration and early-inspiratory (early-I) population of preBotC during inspiration. Therefore the late-E oscillations can be released by either a hypercapnia-evoked activation of chemosensitive RTN/pFRG neurons overcoming this inhibition or a hypoxia-dependent suppression of RTN/pFRG inhibition by BotC-pre-BotC circuits. Our model proposes mechanistic explanations for the emergence of RTN/pFRG oscillations and their interaction with the brain stem respiratory network. The emerging view is that the brain stem respiratory network has rhythmogenic capabilities at multiple hierarchical levels, which allows flexible, state-dependent expression of different rhythmogenic mechanisms under different physiological and metabolic conditions and enables a wide repertoire of respiratory behaviors.

Intermediate and long-term memory are different at the neuronal level in Lymnaea stagnalis (L.)

Both intermediate-term memory (ITM) and long-term memory (LTM) require novel protein synthesis; however, LTM also requires gene transcription. This suggests that the behavioural output of the two processes may be produced differently at the neuronal level. The fresh-water snail, Lymnaea stagnalis, can be operantly conditioned to decrease its rate of aerial respiration and, depending on the training procedure, the memory can last 3h (ITM) or >24h (LTM). RPeD1, one of the 3 interneurons that form the respiratory central pattern generator (CPG) that drives aerial respiration, is necessary for memory formation. By comparing RPeD1’s electrophysiological properties in naïve, ‘ITM-trained’, ‘LTM-trained’ and yoked control snails we discovered that while the behavioural phenotype of memory at 3 and 24h is identical, the situation at the neuronal level is different. When examined 3h after either the ‘ITM’ or ‘LTM’ training procedure RPeD1 activity is significantly depressed. That is, the firing rate, input resistance, excitability and the number of action potential bursts are all significantly decreased. In snails receiving the ITM-training, these changes return to normal 24h post-training. However, in snails receiving the ‘LTM-training’, measured RPeD1 properties (firing rate, excitability, membrane resistance, and the number of action potential bursts fired) are significantly different at 24h than they were at 3h. Additionally, 24h following LTM training RPeD1 appears to be functionally “uncoupled” from its control of the pneumostome as the link between RPeD1 excitation and pneumostome opening is weakened. These data suggest that the behavioural changes occurring during LTM are due to more widespread neuronal reorganization than similar behavioural changes occurring during ITM. Thus ITM and LTM are not just distinct in a chronological and transcriptional manner but are also distinct at the level of neuronal properties.

Synchronization of presynaptic input to motor units of tongue, inspiratory intercostal, and diaphragm muscles

The respiratory central pattern generator distributes rhythmic excitatory input to phrenic, intercostal, and hypoglossal premotor neurons. The degree to which this input shapes motor neuron activity can vary across respiratory muscles and motor neuron pools. We evaluated the extent to which respiratory drive synchronizes the activation of motor unit pairs in tongue (genioglossus, hyoglossus) and chest-wall (diaphragm, external intercostals) muscles using coherence analysis. This is a frequency domain technique, which characterizes the frequency and relative strength of neural inputs that are common to each of the recorded motor units. We also examined coherence across the two tongue muscles, as our previous work shows that, despite being antagonists, they are strongly coactivated during the inspiratory phase, suggesting that excitatory input from the premotor neurons is distributed broadly throughout the hypoglossal motoneuron pool. All motor unit pairs showed highly correlated activity in the low-frequency range (1-8 Hz), reflecting the fundamental respiratory frequency and its harmonics. Coherence of motor unit pairs recorded either within or across the tongue muscles was similar, consistent with broadly distributed premotor input to the hypoglossal motoneuron pool. Interestingly, motor units from diaphragm and external intercostal muscles showed significantly higher coherence across the 10-20-Hz bandwidth than tongue-muscle units. We propose that the lower coherence in tongue-muscle motor units over this range reflects a larger constellation of presynaptic inputs, which collectively lead to a reduction in the coherence between hypoglossal motoneurons in this frequency band. This, in turn, may reflect the relative simplicity of the respiratory drive to the diaphragm and intercostal muscles, compared with the greater diversity of functions fulfilled by muscles of the tongue.

Interacting oscillations in neural control of breathing: modeling and qualitative analysis

In mammalian respiration, late-expiratory (late-E, or pre-inspiratory) oscillations emerge in abdominal motor output with increasing metabolic demands (e.g., during hypercapnia, hypoxia, etc.). These oscillations originate in the retrotrapezoid nucleus/parafacial respiratory group (RTN/pFRG) and couple with the respiratory oscillations generated by the interacting neural populations of the Bötzinger (BötC) and pre-Bötzinger (pre-BötC) complexes, representing the kernel of the respiratory central pattern generator. Recently, we analyzed experimental data on the generation of late-E oscillations and proposed a large-scale computational model that simulates the possible interactions between the BötC/pre-BötC and RTN/pFRG oscillations under different conditions. Here we describe a reduced model that maintains the essential features and architecture of the large-scale model, but relies on simplified activity-based descriptions of neural populations. This simplification allowed us to use methods of dynamical systems theory, such as fast-slow decomposition, bifurcation analysis, and phase plane analysis, to elucidate the mechanisms and dynamics of synchronization between the RTN/pFRG and BötC/pre-BötC oscillations. Three physiologically relevant behaviors have been analyzed: emergence and quantal acceleration of late-E oscillations during hypercapnia, transformation of the late-E activity into a biphasic-E activity during hypercapnic hypoxia, and quantal slowing of BötC/pre-BötC oscillations with the reduction of pre-BötC excitability. Each behavior is elicited by gradual changes in excitatory drives or other model parameters, reflecting specific changes in metabolic and/or physiological conditions. Our results provide important theoretical insights into interactions between RTN/pFRG and BötC/pre-BötC oscillations and the role of these interactions in the control of breathing under different metabolic conditions.

Chronic nicotinic exposure decreases the synaptic strength of nAChR‐mediated glutamatergic input onto neonatal hypoglossal motoneurons

Although nicotinic acetylcholine receptors (nAChRs) are widespread in the CNS their role in the control of breathing is still unclear. This study aims to test the hypothesis that alterations in nAChRs, evoked by chronic nAChR excitation, impairs respiratory‐related neurotransmission onto hypoglossal motoneurons (HMN) that control the tongue. To test our hypothesis we used rhythmic medullary slice preparations and simultaneously recorded from both single respiratory modulated HMN and whole hypoglossal nerve rootlets on day 1–4 in rat pups chronically exposed to nicotine or saline in utero and during the first week of life. Results show that synaptic strength, predominately glutamatergic, was decreased but without changes in the frequency of respiratory rhythm determined by the respiratory central pattern generator (CPG). The average number of spontaneous excitatory postsynaptic potentials (EPSPs), measured during the “expiratory” period, was decreased 42% (P < 0.05), but their magnitude was unchanged. Furthermore, EPSPs could be abolished by bath application of AMPAergic antagonist 6‐cyano‐7‐nitroquinoxaline‐2, 3‐dione (CNQX). These results show that the chronic in utero exposure to nicotine decreases excitatory neurotransmission at HMN synapses and suggests that this change results from frequency of nAChR‐mediated presynpatic release of glutamate. Funded by NIH #1R03HD061613‐01.

Extent of shared presynaptic input to motor units of tongue and inspiratory intercostal muscles

Synchronous activation of motor units is attributed to common presynaptic input to motoneurons, and this is thought to increase the rate of rise of muscle force, and/or facilitate co‐activation of muscles. The inspiratory‐related drive to protrudor and retractor muscles of the tongue is associated with a rapid rate of rise of both EMG activity and force. Here, we examine the magnitude of synchronous discharge of co‐active motor unit pairs within genioglossus (GG, tongue protrudor), hyoglossus (HG, tongue retractor), and inspiratory intercostal (IC, rib cage expander) muscles in urethane‐anesthetized rats. We recorded from pairs of motor units that were spontaneously bursting under command of the respiratory central pattern generator. The strength of synchronous input to each pair was measured with cross correlation followed by the calculation of common input strength (CIS; frequency of synchronous action potentials in excess of those expected due to chance). CIS values for motor units within IC (N=9 pairs, 0.93 +/− 0.27) and tongue muscles (GG, 32 pairs, 0.83 +/− 0.11; HG, 41 pairs 0.67 +/− 0.13) were statistically equivalent. Across‐muscle CIS for tongue muscles was the same as the within muscle values (34 pairs, 0.55 +/− 0.1). CIS across GG and IC (10 pairs, 0.05 +/− 0.1) was less than within muscle values (P<0.01), while CIS between IC and HG was not (28 pairs, 0.29 +/− 0.1, P<0.05). In conclusion, the proportion of shared presynaptic inputs within and across the protrudor and retractor muscles of the rodent tongue is similar. In contrast, shared input to motor units of tongue and rib cage muscles is relatively low, which likely reflects drive from a different pool of premotor neurons.

Activation of the retrotrapezoid nucleus by posterior hypothalamic stimulation.

The retrotrapezoid nucleus (RTN) contains chemically defined neurons (ccRTN neurons) that provide a pH-regulated excitatory drive to the central respiratory pattern generator. Here we test whether ccRTN neurons respond to stimulation of the perifornical hypothalamus (PeF), a region that regulates breathing during sleep, stress and exercise. PeF stimulation with gabazine increased blood pressure, phrenic nerve discharge (PND) and the firing rate of ccRTN neurons in isoflurane-anaesthetized rats. Gabazine produced an approximately parallel upward shift of the steady-state relationship between ccRTN neuron firing rate and end-tidal CO(2), and a similar shift of the relationship between PND and end-tidal CO(2). The central respiratory modulation of ccRTN neurons persisted after gabazine without a change in pattern. Morphine administration typically abolished PND and reduced the discharge rate of most ccRTN neurons (by 25% on average). After morphine administration, PeF stimulation activated the ccRTN neurons normally but PND activation and the central respiratory modulation of the ccRTN neurons were severely attenuated. In the same rat preparation, most (58%) ccRTN neurons expressed c-Fos after exposure to hypercapnic hyperoxia (6-7% end-tidal CO(2); 3.5 h; no hypothalamic stimulation) and 62% expressed c-Fos under hypocapnia (approximately 3% end-tidal CO(2)) after PeF stimulation. Under baseline conditions (approximately 3% end-tidal CO(2), hyperoxia, no PeF stimulation) few (11%) ccRTN neurons expressed c-Fos. In summary, most ccRTN neurons are excited by posterior hypothalamic stimulation while retaining their normal response to CNS acidification. ccRTN neurons probably contribute both to the chemical drive of breathing and to the feed-forward control of breathing associated with emotions and or locomotion.

What to do about apnea of prematurity?

apnea of prematurity and resultant intermittent hypoxic episodes are universal in preterm infants. These events range from presumably benign “periodic” breathing to clinically significant events that may require mechanical ventilation. Persistence of these events is a major factor in prolonging

Abdominal expiratory activity in the rat brainstem–spinal cord in situ: patterns, origins and implications for respiratory rhythm generation

We studied respiratory neural activity generated during expiration. Motoneuronal activity was recorded simultaneously from abdominal (AbN), phrenic (PN), hypoglossal (HN) and central vagus nerves from neonatal and juvenile rats in situ. During eupnoeic activity, low-amplitude post-inspiratory (post-I) discharge was only present in AbN motor outflow. Expression of AbN late-expiratory (late-E) activity, preceding PN bursts, occurred during hypercapnia. Biphasic expiratory (biphasic-E) activity with pre-inspiratory (pre-I) and post-I discharges occurred only during eucapnic anoxia or hypercapnic anoxia. Late-E activity generated during hypercapnia (7-10% CO(2)) was abolished with pontine transections or chemical suppression of retrotrapezoid nucleus/ventrolateral parafacial (RTN/vlPF). AbN late-E activity during hypercapnia is coupled with augmented pre-I discharge in HN, truncated PN burst, and was quiescent during inspiration. Our data suggest that the pons provides a necessary excitatory drive to an additional neural oscillatory mechanism that is only activated under conditions of high respiratory drive to generate late-E activity destined for AbN motoneurones. This mechanism may arise from neurons located in the RTN/vlPF or the latter may relay late-E activity generated elsewhere. We hypothesize that this oscillatory mechanism is not a necessary component of the respiratory central pattern generator but constitutes a defensive mechanism activated under critical metabolic conditions to provide forced expiration and reduced upper airway resistance simultaneously. Possible interactions of this oscillator with components of the brainstem respiratory network are discussed.

The basis of vagal efferent control of heart rate in a neotropical fish,the pacu,Piaractus mesopotamicus

The role of the parasympathetic nervous system, operating via the vagus nerve, in determining heart rate (f(H)) and cardiorespiratory interactions was investigated in the neotropical fish Piaractus mesopotamicus. Motor nuclei of branches of cranial nerves VII, IX and X, supplying respiratory muscles and the heart, have an overlapping distribution in the brainstem, while the Vth motor nucleus is more rostrally located. Respiration-related efferent activity in the cardiac vagus appeared to entrain the heart to ventilation. Peripheral stimulation of the cardiac vagus with short bursts of electrical stimuli entrained the heart at a ratio of 1:1 over a range of frequencies, both below and sometimes above the intrinsic heart rate. Alternatively, at higher bursting frequencies the induced f(H) was slower than the applied stimulus, being recruited by a whole number fraction (1:2 to 1:6) of the stimulus frequency. These effects indicate that respiration-related changes in f(H) in pacu are under direct, beat-to-beat vagal control. Central burst stimulation of respiratory branches of cranial nerves VII, IX and X also entrained the heart, which implies that cardiorespiratory interactions can be generated reflexly. Central stimulation of the Vth cranial nerve was without effect on heart rate, possibly because its central projections do not overlap with cardiac vagal preganglionic neurons in the brainstem. However, bursts of activity recorded from the cardiac vagus were concurrent with bursts in this nerve, suggesting that cardiorespiratory interactions can arise within the CNS, possibly by irradiation from a central respiratory pattern generator, when respiratory drive is high.

Novel neural correlates of operant conditioning in normal and differentially reared Lymnaea

The aerial respiratory behaviour of the mollusc Lymnaea stagnalis is an important homeostatic behaviour that can be operantly conditioned. The central pattern generator underlying this behaviour, as well as motorneurons innervating the respiratory orifice, the pneumostome, have been identified and their activity can be monitored in the semi-intact preparation using electrophysiological recordings. In this study, we used both intact animals and semi-intact preparations to identify novel changes in the respiratory central pattern generator following operant conditioning. In addition, we reared animals in the absence of this respiratory behaviour throughout development, to investigate whether previous experience and activity-dependent plasticity during development are essential to allow neural plasticity in the adult. We found that animals raised normally (allowed to perform aerial respiratory behaviour) exhibited the expected reduction in aerial respiratory behaviour following operant conditioning. Then, using the semi-intact preparation, we identified novel neural changes within the network as a result of the conditioning. These included specific changes at the level of the central pattern generator interneurons, as well as the motor output. In the differentially reared intact animals, there was no behavioural reduction as a result of operant conditioning, although their baseline respiratory behaviour was already significantly reduced as a result of their differential rearing. There were, however, significant differences found in the network parameters in the semi-intact preparation, similar to those observed in normally reared animals. We thus provide evidence for neural plasticity within the network in the absence of significant behavioural changes in differentially reared animals, and show that plasticity was not dependent on previous activity of the network during development.

Swallow Pattern Generator Reconfiguration of the Respiratory Neural Network

Abstract When the nasal and oral passages converge and a bolus enters the pharynx, it is critical that breathing and swallow motor patterns become integrated to allow safe passage of the bolus through the pharynx. Breathing patterns must be reconfigured to inhibit inspiration, and upper airway muscle activity must be recruited and reconfigured to close the glottis and laryngeal vestibule, invert the epiglottis, and ultimately protect the lower airways. Failure to close and protect the glottal opening to the lower airways, or loss of the integration and coordination of swallow and breathing, increases the risk of penetration or aspiration. A neural swallow central pattern generator (CPG) controls the pharyngeal swallow phase and is located in the medulla. We propose that this swallow CPG is functionally organized in a holarchical behavioral control assembly (BCA) and is recruited with pharyngeal swallow. The swallow BCA holon reconfigures the respiratory CPG to produce the stereotypical swallow breathing pattern, consisting of swallow apnea during swallowing followed by prolongation of expiration following swallow. The timing of swallow apnea and the duration of expiration is a function of the presence of the bolus in the pharynx, size of the bolus, bolus consistency, breath cycle, ventilatory state and disease.

Calcium-activated nonspecific cation current and synaptic depression promote network-dependent burst oscillations

Central pattern generators (CPGs) produce neural-motor rhythms that often depend on specialized cellular or synaptic properties such as pacemaker neurons or alternating phases of synaptic inhibition. Motivated by experimental evidence suggesting that activity in the mammalian respiratory CPG, the preBötzinger complex, does not require either of these components, we present and analyze a mathematical model demonstrating an unconventional mechanism of rhythm generation in which glutamatergic synapses and the short-term depression of excitatory transmission play key rhythmogenic roles. Recurrent synaptic excitation triggers postsynaptic Ca(2+)-activated nonspecific cation current (I(CAN)) to initiate a network-wide burst. Robust depolarization due to I(CAN) also causes voltage-dependent spike inactivation, which diminishes recurrent excitation and thus attenuates postsynaptic Ca(2+) accumulation. Consequently, activity-dependent outward currents-produced by Na/K ATPase pumps or other ionic mechanisms-can terminate the burst and cause a transient quiescent state in the network. The recovery of sporadic spiking activity rekindles excitatory interactions and initiates a new cycle. Because synaptic inputs gate postsynaptic burst-generating conductances, this rhythm-generating mechanism represents a new paradigm that can be dubbed a 'group pacemaker' in which the basic rhythmogenic unit encompasses a fully interdependent ensemble of synaptic and intrinsic components. This conceptual framework should be considered as an alternative to traditional models when analyzing CPGs for which mechanistic details have not yet been elucidated.

Retrotrapezoid nucleus, respiratory chemosensitivity and breathing automaticity.

Breathing automaticity and CO(2) regulation are inseparable neural processes. The retrotrapezoid nucleus (RTN), a group of glutamatergic neurons that express the transcription factor Phox2b, may be a crucial nodal point through which breathing automaticity is regulated to maintain CO(2) constant. This review updates the analysis presented in prior publications. Additional evidence that RTN neurons have central respiratory chemoreceptor properties is presented, but this is only one of many factors that determine their activity. The RTN is also regulated by powerful inputs from the carotid bodies and, at least in the adult, by many other synaptic inputs. We also analyze how RTN neurons may control the activity of the downstream central respiratory pattern generator. Specifically, we review the evidence which suggests that RTN neurons (a) innervate the entire ventral respiratory column and (b) control both inspiration and expiration. Finally, we argue that the RTN neurons are the adult form of the parafacial respiratory group in neonate rats.

GABAA- and AMPA-like receptors modulate the activity of an identified neuron within the central pattern generator of the pond snail Lymnaea stagnalis

To examine the neurochemistry underlying the firing of the RPeD1 neuron in the respiratory central pattern generator of the pond snail, Lymnaea stagnalis, we examined electrophysiologically and pharmacologically either "active" or "silent" preparations by intracellular recording and pharmacology. GABA inhibited electrical firing by hyperpolarizing RPeD1, while picrotoxin, an antagonist of GABA(A) receptors, excited silent cells and reversed GABA-induced inhibition. Action potential activity was terminated by 1 mM glutamate (Glu) while silent cells were depolarized by the GluR agonists, AMPA, and NMDA. Kainate exerted a complex triphasic effect on membrane potential. However, only bath application of AMPA desensitized the firing. These data indicate that GABA inhibits RPeD1 via activation of GABA(A) receptors, while Glu stimulates the neuron by activating AMPA-sensitive GluRs.

Multiple Rhythmic States in a Model of the Respiratory Central Pattern Generator

The three-phase respiratory pattern observed during normal breathing changes with alterations in metabolic or physiological conditions. A recent study using in situ perfused rat brain preparations demonstrated a reorganization of the respiratory pattern with sequential reduction of the brain stem respiratory network. Specifically, with removal of the pons, the normal three-phase pattern transformed to a two-phase inspiratory-expiratory pattern and, with more caudal transections, to one-phase, intrinsically generated inspiratory oscillations. A minimal neural network proposed to reproduce these transformations includes 1) a ringlike mutually inhibitory network composed of the postinspiratory, augmenting expiratory, and early-inspiratory neurons and 2) an excitatory preinspiratory neuron, with persistent sodium current (I(NaP))-dependent intrinsic bursting properties, that dynamically participates in the expiratory-inspiratory phase transition and inspiratory phase generation. We used activity-based single-neuron models and applied numerical simulations, bifurcation methods, and fast-slow decomposition to describe the behavior of this network in the functional states corresponding to the three-, two-, and one-phase oscillatory regimes, as well as to analyze the transitions between states and between respiratory phases within each state. We demonstrate that, although I(NaP) is not necessary for the generation of three- and two-phase oscillations, it contributes to control of the oscillation period in each state. We also show that the transitions between states can be produced by progressive changes of drives to particular neurons and proceed through intermediate regimes, featuring high-amplitude late-expiratory and biphasic-expiratory activities or ectopic burst generation. Our results provide important insights for understanding the state-dependent mechanisms for respiratory rhythm generation and control.

Hypoxia-induced modulation of the respiratory CPG

Despite recent advances in our understanding of the neural control of breathing, the precise cellular, synaptic, and molecular mechanisms underlying the generation and modulation of respiratory rhythm remain largely unknown. This lack of fundamental knowledge in the field of neural control of respiration is likely due to the complexity of the mammalian brain where synaptic connectivity between central respiratory neurons, motor neurons and their peripheral counterparts cannot be mapped reliably. We have therefore developed an invertebrate model system wherein the essential elements of the central pattern generator (CPG), the motor neurons and the peripheral chemosensory cells involved in respiratory control have been worked out both in vivo and in vitro. We discuss our recent identification of peripheral, hypoxia sensitive chemoreceptor elements in a sensory organ of the pulmonate freshwater pond snail Lymnaea stagnalis, which provide an excitatory drive to the respiratory CPG neuron RPeD1 via direct chemical synaptic connections. Further studies using this unique invertebrate model system may reveal highly conserved principles of CPG neuromodulation that will remain relevant to more complex mammalian systems.

A non-selective cationic channel regulating respiratory CPG activity of Lymnaea stagnalis

Event Abstract Back to Event A non-selective cationic channel regulating respiratory CPG activity of Lymnaea stagnalis Tom Z. Lu1* and Zhong-Ping Feng1 1 University of Toronto, Department of Physiology, Faculty of Medicine, Canada NALCN (Na+ leak current, nonselective) is a newly described cation channel characterized as voltage-independent, nonselective, and noninactivating. The channels conduct Na+ current at rest, and potentially contribute toward neuronal excitability and synaptic transmission. Targeted deletion of NALCN gene in mice resulted in a fatal postnatal phenotype characterized by an abnormal respiratory rhythmic activity. However, the fundamental question of whether the NALCN channels is involved in generating rhythmic activity of respiratory central pattern generator (rCPG) neurons has not been addressed. Currently, a lack of a suitable model hinders directly addressing this question in mature respiratory network activity of mammalian system. Lymnaea stagnalis is a bimodal breather: its respiratory activity is controlled by a well-described CPG network consisting of 3 identified neurons. Thus L. stagnalis serves as a convenient model to study the mechanisms of respiratory rhythm in identified CPG network. In this study, we determined the role of U-type channel (GenBank: AF484086, AF484085; an NALCN orthologue) in the membrane excitability of an identified rCPG neuron in adult L. stagnalis. We first determined the ganglial expression of the U-type gene in the snail, and subsequently used a dsRNA gene silencing approach to acutely knockdown the expression of U-type channel gene, using real-time PCR analysis. Respiratory behavioural study showed a reduction of basal aerial respiratory activity following partial acute U-type gene knockdown. Intracellular sharp recordings from the pacemaker rCPG neuron, RPeD1, in isolated ganglial preparation showed a decrease in action potential frequency and resting membrane potential in the U-type dsRNA treatment group. Whole-cell (ruptured) voltage-clamp recordings showed U-type channel dsRNA treated cells exhibited a reduced inward hyperpolarizing cation current component likely contributed by the U-type channel. Taken together, our results provided the first evidence of the functional channel expression in the respiratory CPG neurons of L. stagnalis and the involvement of the channel in the respiratory rhythmic activity via regulation of intrinsic membrane properties. Conference: B.R.A.I.N. platform in Physiology poster day 2009, Toronto, ON, Canada, 16 Dec - 16 Dec, 2009. Presentation Type: Poster Presentation Topic: Poster presentations Citation: Lu TZ and Feng Z (2009). A non-selective cationic channel regulating respiratory CPG activity of Lymnaea stagnalis. Front. Neurosci. Conference Abstract: B.R.A.I.N. platform in Physiology poster day 2009. doi: 10.3389/conf.neuro.03.2009.17.037 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 17 Dec 2009; Published Online: 17 Dec 2009. * Correspondence: Tom Z Lu, University of Toronto, Department of Physiology, Faculty of Medicine, Toronto, Canada, 2008tomlu@gmail.com Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Tom Z Lu Zhong-Ping Feng Google Tom Z Lu Zhong-Ping Feng Google Scholar Tom Z Lu Zhong-Ping Feng PubMed Tom Z Lu Zhong-Ping Feng Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.