Postinspiratory Activity Research Articles

Electrophysiological Properties of Ventral Medullary Respiratory Neurons in Rats Submitted to Chronic Intermittent Hypoxia

The central network controlling breathing exhibits plasticity in response to environmental challenges, such as chronic intermittent hypoxia (CIH). For a better understanding of the effects of CIH on the central generation of breathing, we combined single cell qRT-PCR, nerve and whole cell patch clamp recordings of Bötzinger Complex (BötC) and pre-Bötzinger Complex (pre-BötC) respiratory neurons in in situ preparations from control and CIH rats. In the BötC, we recorded 32 augmenting-expiratory (aug-E; glicinergic) and 24 post-inspiratory (post-I; 19 glicinergic and 5 glutamatergic) neurons, which present distinct intrinsic electrophysiological properties. CIH decreased the superior laryngeal nerve (SLN) post-inspiratory activity, by increasing potassium-mediated leak current in the post-I neurons, and increased the duration of expiration by changes in the aug-E neurons. In the pre-BötC, we recorded four types of neurons: pre-inspiratory (pre-I; 15 glutamatergic, 9 GABAergic and 11 glicinergic), early-inspiratory (19 glicinergic neurons), ramp-inspiratory (13 glicinergic and 9 GABAergic) and late-inspiratory (7 GABAergic and 9 glicinergic). CIH enhanced SLN pre-I activity, by increasing the inward riluzole-sensitive persistent sodium current in the intrinsically bursting glutamatergic pre-I neurons. These data show that CIH triggers a reconfiguration of synaptic interaction within the respiratory network in rats, which seems to be an adaptive process required for appropriated pulmonary ventilation and regulation of upper airway resistance. Support: FAPESP, CAPES and CNPQ.

Chapter 12 - Bidirectional Plasticity of Pontine Pneumotaxic Postinspiratory Drive: Implication for a Pontomedullary Respiratory Central Pattern Generator

The "pneumotaxic center" in the rostral dorsolateral pons as delineated by Lumsden nine decades ago is known to play an important role in promoting the inspiratory off-switch (IOS) for inspiratory-expiratory phase transition as a fail-safe mechanism for preventing apneusis in the absence of vagal input. Traditionally, the pontine pneumotaxic mechanism has been thought to contribute a tonic descending input that lowers the IOS threshold in medullary respiratory central pattern generator (rCPG) circuits, but otherwise does not constitute part of the rCPG. Recent evidence indicates that descending input from the Kölliker-Fuse nucleus (KFN) within the pneumotaxic center is essential for gating the postinspiratory phase of the three-phase respiratory rhythm to control the IOS in vagotomized animals. A critical question arising is whether such a descending pneumotaxic input from KFN that drives postinspiratory activity is tonic (null hypothesis) or rhythmic with postinspiratory phase modulation (alternative hypothesis). Here, we show that multifarious evidence reported in the literature collectively indicates that the descending pneumotaxic input may exhibit NMDA receptor-dependent short-term plasticity in the form of a biphasic neural differentiator that bidirectionally and phase-selectively modulates postinspiratory phase duration in response to vagal and peripheral chemoreceptor inputs independent of the responses in inspiratory and late-expiratory activities. The phase-selectivity property of the descending pneumotaxic input implicates a population of pontine early-expiratory (postinspiratory/expiratory-decrementing) neurons as the most likely neural correlate of the pneumotaxic mechanism that drives post-I activity, suggesting that the pontine pneumotaxic mechanism may be an integral part of a pontomedullary rCPG that underlies the three-phase respiratory rhythm.

Chapter 7 - The physiological significance of postinspiration in respiratory control

The term postinspiration is commonly used in the scientific literature concerned with neural generation and the control of breathing movements. Because postinspiration belongs functionally to the mechanical act of expiration, the physiological significance of postinspiration as a distinct phase of the breathing cycle is often underappreciated. The present review will give an overview of the physiological significance of postinspiratory motor activity in laryngeal adductor (constrictor) muscles and the crural diaphragm. The functional importance of postinspiratory motor activity is discussed for the eupneic respiratory cycle, and for various protective respiratory reflex mediations (e.g., sneeze, cough, and breath-hold). Also, the implications of recruited postinspiratory activity during nonventilatory behaviors such as vocalization, swallowing, or vomiting are underpinned. Finally, we describe the impact of absence or malfunction of postinspiratory motor function in neurological diseases.

Characterization of the relationship between inspiratory neural discharges and the micturition reflex in decerebrate and urethane‐anesthetized adult in vivo rat

Previous studies have characterized respiratory neural outputs during the micturition reflex (MR). However, these studies primarily were conducted in cats, and it is not known if the relationship seen in the cat model is maintained in rodent models. To begin to address this issue, we recorded bladder pressure and phrenic (Phr), hypoglossal (XII) and pudendal motor nerve activity in decerebrate or urethane‐anesthetized vagi‐intact adult rats during passive bladder filling and active bladder contraction. Spontaneous MRs were elicited by slow infusion of saline into the bladder. The data show increased Phr post‐inspiratory activity and a transient increase followed by a transient decrease in burst frequency (due to changes in TE). Small XII neural discharges were observed during expiration while the bladder actively contracted. In contrast to cat data, no changes in Phr or XII amplitude were observed. Finally, while severe hypercapnia and hypocapnia affected respiratory patterns, they did not affect the MR. We conclude that, while there are some similarities to the cat model, a different relationship between respiration and micturition occurs in rat. Supported by HL63175

Expiratory-modulated laryngeal motoneurons exhibit a hyperpolarization preceding depolarization during superior laryngeal nerve stimulation in the in vivo adult rat

Swallowing requires the sequential activation of tongue, pharyngeal and esophageal muscles to propel the food bolus towards the stomach. Aspiration during swallow is prevented by adduction of the vocal cords during the oropharyngeal phase. Expiratory-modulated laryngeal motoneurons (ELM) exhibit a burst of action potentials during swallows elicited by electrical stimulation of the superior laryngeal nerve (SLN). Here we sought to investigate changes in membrane potential in ELM during superior laryngeal nerve stimulation in the anaesthetised, in vivo adult rat preparation. Intracellular recordings of ELM in the caudal nucleus ambiguus (identified by antidromic activation from the recurrent laryngeal nerve) demonstrated that ELM bursting activity following SLN stimulation is associated with a depolarization that is preceded by a small hyperpolarization. During spontaneous ELM bursts, the preceding hyperpolarization separated the bursting activity from its usual post-inspiratory activity. These findings demonstrate that the in vivo adult rat preparation is suitable for the study of swallow-related activity in laryngeal motoneurons.

Neuronal Mechanisms Underlying the Laryngeal Adductor Reflex

Electromyographic studies of the laryngeal adductor reflex, glottal closure occurring in response to laryngeal stimulation, have demonstrated an early ipsilateral response (R1) and a late bilateral response (R2). To better define the physiologic properties of these responses, we recorded responses from expiratory laryngeal motoneurons (ELMs) in rats during stimulation of the superior laryngeal nerve (SLN). Single unit extracellular recordings were obtained from 5 ELMs, identified by their antidromic responses to recurrent laryngeal nerve stimulation and postinspiratory firing pattern, in 4 Sprague-Dawley rats. Unilateral stimulation of the SLN (at 20 Hz) stopped both phrenic nerve inspiratory activity and ELM postinspiratory activity. However, the ELMs displayed robust tonic firing, consisting of non-respiratory burst activity and single action potentials. The single action potentials were identified as short-latency ones (5 to 10 ms) activated by ipsilateral SLN stimulation, with an occurrence rate of 90%, and long-latency ones (20 to 50 ms) activated by bilateral SLN stimulation, with occurrence rates of 47% on the ipsilateral side and 58% on the contralateral side. The R1 response appears to be the result of the short-latency action potentials, orthodromically activated by ipsilateral stimulation of the SLN. The R2 response is likely to be a result of the long-latency action potentials that can be recorded from ELMs on both sides.

Computational modelling of 5-HT receptor-mediated reorganization of the brainstem respiratory network

Brainstem respiratory neurons express the glycine α(3) receptor (Glyα(3) R), which is a target of modulation by several serotonin (5-HT) receptor agonists. Application of the 5-HT(1A) receptor (5-HT(1A) R) agonist 8-OH-DPAT was shown (i) to depress cellular cAMP, leading to dephosphorylation of Glyα(3) R and augmentation of postsynaptic inhibition of neurons expressing Glyα(3) R (Manzke etal., 2010) and (ii) to hyperpolarize respiratory neurons through 5-HT-activated potassium channels. These processes counteract opioid-induced depression and restore breathing from apnoeas often accompanying pharmacotherapy of pain. The effect is postulated to rely on the enhanced Glyα(3) R-mediated inhibition of inhibitory neurons causing disinhibition of their target neurons. To evaluate this proposal and investigate the neural mechanisms involved, an established computational model of the brainstem respiratory network (Smith etal., 2007), was extended by (i) incorporating distinct subpopulations of inhibitory neurons (glycinergic and GABAergic) and their synaptic interconnections within the Bötzinger and pre-Bötzinger complexes and (ii) assigning the 5-HT(1A) R-Glyα(3) R complex to some of these inhibitory neuron types in the network. The modified model was used to simulate the effects of 8-OH-DPAT on the respiratory pattern and was able to realistically reproduce a number of experimentally observed responses, including the shift in the onset of post-inspiratory activity to inspiration and conversion of the eupnoeic three-phase rhythmic pattern into a two-phase pattern lacking the post-inspiratory phase. The model shows how 5-HT(1A) R activation can produce a disinhibition of inspiratory neurons, leading to the recovery of respiratory rhythm from opioid-induced apnoeas.

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.

A μ-opioid receptor agonist DAMGO induces rapid breathing in the arterially perfused in situ preparation of rat

Ventilatory responses to opioids are complex and not yet fully understood. We evaluated concentration-dependent effects of a selective μ-opioid receptor agonist [D-Ala(2),N-Me-Phe(4),Gly(5)-ol]-enkephalin (DAMGO) on respiratory output in the arterially perfused in situ rat preparation, which preserves the integrity of the ponto-medullary respiratory network. DAMGO (300-3400 nM) was added accumulatively to the perfusate. DAMGO increased inspiratory time and diminished central vagal post-inspiratory activity. At 300-500 nM DAMGO caused rapid breathing with shortening of expiratory time. The change of breathing pattern occurred within a single breath. Bilateral vagotomy did not affect the change in respiratory pattern, suggesting that it was of central origin. Additional DAMGO up to 1800 nM did not affect the rapid breathing pattern, and further elevated concentrations (up to 3400 nM) caused inconsistent results. Since the rapid breathing pattern was associated with the obliteration of vagal post-inspiratory activity, we conclude that DAMGO reconfigures the respiratory output to an inspiratory phase-dominant, rapidly alternating inspiration-expiration pattern.

Midbrain and medullary control of postinspiratory activity of the crural and costal diaphragm in vivo

Studies on brain stem respiratory neurons suggest that eupnea consists of three phases: inspiration, postinspiration, and expiration. However, it is not well understood how postinspiration is organized in the diaphragm, i.e., whether postinspiration differs in the crural and costal segments of the diaphragm and what the influence is of postinspiratory neurons on diaphragm function during eupnea. In this in vivo study we investigated the postinspiratory activity of the two diaphragm segments during eupnea and the changes in diaphragm function following modulation of eupnea. Postinspiratory neurons in the medulla were stereotaxically localized extracellularly and neurochemically stimulated. We used three types of preparations: precollicularly decerebrated unanesthetized cats and rats and anesthetized rats. In all preparations, during eupnea, postinspiratory activity was found in the crural but not in the costal diaphragm. When eupnea was discontinued in decerebrate cats in which stimulation in the nucleus retroambiguus induced activation of laryngeal or abdominal muscles, all postinspiratory activity in the crural diaphragm was abolished. In decerebrate rats, stimulation of the midbrain periaqueductal gray abolished postinspiration in the crural diaphragm but induced activation in the costal diaphragm. In anesthetized rats, stimulation of medullary postinspiratory neurons abolished the postinspiratory activity of the crural diaphragm. Vagal nerve stimulation in these rats increased the intensity of postinspiratory neuronal discharge in the solitary nucleus, leading to decreased activity of the crural diaphragm. These data demonstrate that three-phase breathing in the crural diaphragm during eupnea exists in vivo and that postinspiratory neurons have an inhibitory effect on crural diaphragm function.

Respiratory Muscle Activity Related to Flow and Lung Volume in Preterm Infants Compared With Term Infants

Infants with chronic lung disease (CLD) have a capacity to maintain functional lung volume despite alterations to their lung mechanics. We hypothesize that they achieve this by altering breathing patterns and dynamic elevation of lung volume, leading to differences in the relationship between respiratory muscle activity, flow and lung volume. Lung function and transcutaneous electromyography of the respiratory muscles (rEMG) were measured in 20 infants with CLD and in 39 healthy age-matched controls during quiet sleep. We compared coefficient of variations (CVs) of rEMG and the temporal relationship of rEMG variables, to flow and lung volume [functional residual capacity (FRC)] between these groups. The time between the start of inspiratory muscle activity and the resulting flow (tria)--in relation to respiratory cycle time--was significantly longer in infants with CLD. Although FRC had similar associations with tria and postinspiratory activity (corrected for respiratory cycle time), the CV of the diaphragmatic rEMG was lower in CLD infants (22.6 versus 31.0%, p = 0.030). The temporal relationship of rEMG to flow and FRC and the loss of adaptive variability provide additional information on coping mechanisms in infants with CLD. This technique could be used for noninvasive bedside monitoring of CLD.

Essential Role of Phox2b-Expressing Ventrolateral Brainstem Neurons in the Chemosensory Control of Inspiration and Expiration

Phox2b-expressing neurons of the retrotrapezoid nucleus (RTN), located in the ventrolateral brainstem, are sensitive to changes in PCO(2)/pH, have excitatory projections to the central respiratory rhythm/pattern generator, and their activation enhances central respiratory drive. Using in vivo (conscious and anesthetized rats) and in situ (arterially perfused rat brainstem-spinal cord preparations) models, we evaluated the functional significance of this neuronal population for both resting respiratory activity and the CO(2)-evoked respiratory responses by reversibly inhibiting these neurons using the insect peptide allatostatin following transduction with a lentiviral construct to express the G-protein-coupled Drosophila allatostatin receptor. Selective inhibition of the Phox2b-expressing neurons in the ventrolateral brainstem, including the RTN, using allatostatin was without effect on resting respiratory activity in conscious rats, but decreased the amplitude of the phrenic nerve discharge in anesthetized rats and the in situ rat preparations. Postinspiratory activity was also reduced in situ. In the absence or presence of the peripheral chemoreceptor input, inhibiting the Phox2b-expressing neurons during hypercapnia abolished the CO(2)-evoked abdominal expiratory activity in anesthetized rats and in situ preparations. Inspiratory responses evoked by rising levels of CO(2) in the breathing air were also reduced in anesthetized rats with denervated carotid bodies and conscious rats with peripheral chemoreceptors intact (by 28% and 60%, respectively). These data indicate a crucial dependence of central expiratory drive upon Phox2b-expressing neurons of the ventrolateral brainstem and support the hypothesis that these neurons contribute in a significant manner to CO(2)-evoked increases of inspiratory activity.

Sympathetic overactivity coupled with active expiration in rats submitted to chronic intermittent hypoxia

It is well known that the respiration modulates sympathetic outflow in basal conditions. Recordings of sympathetic nerve activity demonstrated that central respiratory activity produces rhythmical oscillations in sympathetic discharge that appear mainly during inspiratory phase. This led us to hypothesize that changes in the mechanisms regulating the central entrainment between respiratory and sympathetic activities may contribute to sympathetic overactivity and hypertension. This issue was addressed using rats submitted to chronic intermittent hypoxia (CIH), in which we evaluated whether or not the sympathetic overactivity and hypertension observed in these animals were linked to changes in respiratory pattern. We verified that under baseline conditions, CIH rats exhibited a reduction in post-inspiratory activity of vagus nerve and an enhanced late-expiratory activity in abdominal motor nerve. As a consequence of this altered expiratory pattern, we observed that CIH rats showed an additional burst in sympathetic activity phase-locked with the enhanced late-E expiratory activity. These findings pointed out that the entrainment between pontine-medullary expiratory and sympathetic neurons of CIH rats is strengthened, indicating for the first time in this experimental model that changes in the coupling of respiratory and sympathetic activities may contribute to hypertension. Subsequent studies performed in other models of hypertension also demonstrated similar changes, supporting the concept that alterations in central mechanisms of respiratory-sympathetic coupling is a novel and important mechanism to be considered in the pathogenesis of hypertension.

Glutamatergic Antagonism in the NTS Decreases Post-Inspiratory Drive and Changes Phrenic and Sympathetic Coupling During Chemoreflex Activation

For a better understanding of the processing at the nucleus tractus solitarius (NTS) level of the autonomic and respiratory responses to peripheral chemoreceptor activation, herein we evaluated the role of glutamatergic neurotransmission in the intermediate (iNTS) and caudal NTS (cNTS) on baseline respiratory parameters and on chemoreflex-evoked responses using the in situ working heart-brain stem preparation (WHBP). The activities of phrenic (PND), cervical vagus (cVNA), and thoracic sympathetic (tSNA) nerves were recorded before and after bilateral microinjections of kynurenic acid (Kyn, 5 nmol/20 nl) into iNTS, cNTS, or both simultaneously. In WHBP, baseline sympathetic discharge markedly correlated with phrenic bursts (inspiration). However, most of sympathoexcitation elicited by chemoreflex activation occurred during expiration. Kyn microinjected into iNTS or into cNTS decreased the postinspiratory component of cVNA and increased the duration and frequency of PND. Kyn into iNTS produced no changes in sympathoexcitatory and tachypneic responses to peripheral chemoreflex activation, whereas into cNTS, a reduction of the sympathoexcitation, but not of the tachypnea, was observed. The pattern of phrenic and sympathetic coupling during the chemoreflex activation was an inspiratory-related rather than an expiratory-related sympathoexcitation. Kyn simultaneously into iNTS and cNTS produced a greater decrease in postinspiratory component of cVNA and increase in frequency and duration of PND and abolished the respiratory and autonomic responses to chemoreflex activation. The data show that glutamatergic neurotransmission in the iNTS and cNTS plays a tonic role on the baseline respiratory rhythm, contributes to the postinspiratory activity, and is essential to expiratory-related sympathoexcitation observed during chemoreflex activation.

Somatostatin selectively ablates post-inspiratory activity after injection into the Bötzinger complex

Somatostatin (SST) neurons in the ventral respiratory column (VRC) are essential for the generation of normal breathing. Little is known about the neuromodulatory role of SST on ventral respiratory neurons other than that local administration induces apnoea. Here, we describe the cardiorespiratory effects of microinjecting SST into the preBötzinger and Bötzinger complexes which together elaborate a normal inspiratory augmenting and expiratory respiratory pattern, and on spinally projecting respiratory subnuclei (rostral ventral respiratory group; rVRG). Microinjections (20–50 nl) of SST (0.15, 0.45, 1.5 mM) were made into respiratory subnuclei of urethane-anaesthetized, paralysed, vagotomized and artificially ventilated Sprague–Dawley rats (n=46). Unilateral microinjection of SST into the Bötzinger complex converted the augmenting activity of phrenic nerve discharge into a square-wave apneustic pattern associated with a lengthening of inspiratory period and shortening of expiratory time. Following bilateral microinjection the apneusis became pronounced and was associated with a dramatic variability in inspiratory duration. Microinjection of SST into the Bötzinger complex also abolished the post-inspiratory (post-I) motor activity normally observed in vagal and sympathetic nerves. In the preBötzinger complex SST caused bradypnoea and with increasing dose, apnoea. In the rVRG SST reduced phrenic nerve amplitude, eventually causing apnoea. In conclusion, SST powerfully inhibits respiratory neurons throughout the VRC. Of particular interest is the finding that chemical inhibition of the Bötzinger complex with SST ablates the post-I activity that is normally seen in respiratory activity and leads to apneusis. This loss of post-I activity is a unique feature of inhibition with SST and is not seen following inhibition with other agents such as galanin, GABA and endomorphin. The effect seen on post-I activity is similar to the effect of inhibiting the Kölliker–Fuse nucleus in the pons. The mechanism by which SST exerts this effect on Bötzinger neurons remains to be determined.

Chronic blockade of 5‐HT and NE reuptake alters the timing and patterning of phrenic nerve discharge during eupnea, but not gasping, in arterially‐perfused adult mouse

Previous in vivo and in vitro studies have demonstrated that a long‐term increase in either 5‐HT or NE exerts complex modulatory effects on central respiratory neural control, resulting in a depression of respiratory motor output. The influence of excess endogenous 5‐HT and NE chronically on respiratory motor output, however, requires clarification. Therefore, in an arterially‐perfused adult mouse preparation, we examined phrenic nerve discharge following intraperitoneal (ip) administration for 28‐d of the 5‐HT and NE reuptake inhibitor venlafaxine hydrochloride (VHCL). The results from these experiments were compared with those obtained from a separate group of mice that received ip injections of 0.9% NaCl (vehicle). We found that in mice receiving VHCL, basal phrenic burst frequency was slower by ~45% (P<0.05), with the effects on TI and TE being somewhat variable. Further, post‐inspiratory activity and spurious expiratory bursts were noted. In response to hypoxia/ischemia, we found that both groups exhibited similar acute hypoxic ventilatory responses and ischemia‐induced gasping responses, with gasps exhibiting similar timing and patterning characteristics. These results demonstrate that chronic excess endogenous 5‐HT and NE depress basal respiratory motor output, but exert minimal effects on hypoxic responses. Supported by NS045321

Abdominal expiratory muscle activity in anesthetized vagotomized neonatal rats

The pattern of respiratory activity in abdominal muscles was studied in anesthetized, spontaneously breathing, vagotomized neonatal rats at postnatal days 0-3. Anesthesia (2.0% isoflurane, 50% O(2)) depressed breathing and resulted in hypercapnia. Under this condition, abdominal muscles showed discharge late in the expiratory phase (E2 activity) in most rats. As the depth of anesthesia decreased, the amplitude of discharges in the diaphragm and abdominal muscles increased. A small additional burst frequently occurred in abdominal muscles just after the termination of diaphragmatic inspiratory activity (E1 or postinspiratory activity). Since this E1 activity is not often observed in adult rats, the abdominal respiratory pattern likely changes during postnatal development. Anoxia-induced gasping after periodic expiratory activity without inspiratory activity, and in most rats, abdominal expiratory activity disappeared before terminal apnea. These results suggest that a biphasic abdominal motor pattern (a combination of E2 and E1 activity) is a characteristic of vagotomized neonatal rats during normal respiration.

Postnatal emergence of synaptic plasticity associated with dynamic adaptation of the respiratory motor pattern

The shape of the three-phase respiratory motor pattern (inspiration, postinspiration, late expiration) is controlled by a central pattern generator (CPG) located in the ponto-medullary brainstem. Synaptic interactions between and within specific sub-compartments of the CPG are subject of intensive research. This review addresses the neural control of postinspiratory activity as the essential determinant of inspiratory/expiratory phase duration. The generation of the postinspiratory phase depends on synaptic interaction between neurones of the nucleus tractus solitarii (NTS), which relay afferent inputs from pulmonary stretch receptors, and the pontine Kölliker-Fuse nucleus (KF) as integral parts of the CPG. Both regions undergo significant changes during the first three postnatal weeks in rodents. Developmental changes in glutamatergic synaptic functions and its modulation by brain-derived neurotrophic factor may have implications in synaptic plasticity within the NTS/KF axis. We propose that dependent on these developmental changes, the CPG becomes permissive for short- and long-term plasticity associated with environmental, metabolic and behavioural adaptation of the breathing pattern.

GABAA mediated inhibition and post-inspiratory pattern of laryngeal constrictor motoneurons in rat

Laryngeal constrictor motoneurons (LCMN) are activated during post-inspiration and act to slow expiratory airflow. However, little is known about how this phasic activity is generated. Here, we investigated the electrophysiological responses of identified LCMN to local application of GABA and bicuculline methiodide (BIC) in 14 anaesthetised Sprague-Dawley rats. During extracellular recordings, GABA iontophoresis (0.5M) strongly inhibited LCMN (n=6). Interestingly, BIC iontophoresis (5 mM) reduced, rather than increased, LCMN post-inspiratory activity (5 out of 6). Furthermore, intracellular recording revealed that BIC reduced not only the hyperpolarisation of the LCMN during inspiration (2.5+/-1.4 mV before and 1.5+/-0.4 mV after the BIC, P=0.05, n=5), but also the depolarisation during post-inspiration (3.0+/-1.3 mV before and 1.6+/-0.4 mV after the BIC, P=0.02, n=5). Our results demonstrate for the first time that the inspiratory inhibition of LCMN is primarily mediated by GABA(A) receptors. A possible involvement of a post-inhibitory rebound mechanism is discussed to explain how blockade of an inspiratory inhibition would affect LCMN excitability during post-inspiration.

Activation of ionotropic excitatory amino acid (EAA) receptors modulates both eupnea and gasping in the arterially‐perfused adult rat preparation

Previous work from our laboratory in an arterially‐perfused adult rat preparation has shown that blockade of ionotropic EAA receptors reduces or abolishes basal phrenic nerve discharge and markedly enhances inspiratory neural network complexity. Further, we have demonstrated that during EAA receptor blockade, gasping can still be elicited. The effect of activation of ionotropic glutamate receptors on phrenic motor discharge requires clarification. We examined the effects of perfusion with the glutamate analog DLH on phrenic burst timing and inspiratory neural network complexity in the arterially‐perfused adult rat preparation. DLH (500 μM, n=8) transiently increased phrenic burst frequency by 41% (P=0.005) mediated by decreasing the duration between bursts by 55% (p<0.001), followed by a 36% increased stabilization of burst frequency (P=0.074). Post‐inspiratory activity was also increased. DLH decreased the duration of the peripheral chemoreflex from 23.8±1.7 to 13.2±1.4 s (P<0.001), the duration of ischemia‐induced gasping from 58.6±5.3 to 33.4±2.3 s (P<0.001), and the onset latency to gasping by 79% (P<0.001). These findings support a role for ionotropic EAA neurotransmission in establishing the temporal dynamics of basal phrenic nerve discharge and suggest that it also has a role in determining the onset and duration of the peripheral chemoreflex and gasping. Supported by NS045321 and NS049310