Respiratory-related Unit Research Articles

Rubral modulation of breathing.

What is the topic of this review? Rubral modulation of pontomedullary respiratory rhythm and pattern generating circuitry powerfully contributes to regulation of breathing. What advances does it highlight? Studies have demonstrated extensive rubromedullary and rubrospinal projections to zones generating and organizing the respiratory rhythm and pattern. Rubral modulation of respiratory output effects inspiratory expiratory phase transitions with stimulation generating inhibitory or excitatory responses of medullary inspiratory and expiratory units. The red nucleus mediates hypoxic ventilatory depression, integrates respiratory output with oromotor and locomotor activity, and modulates respiratory output during noxious stimulation. Although normal triphasic eupnoea can be produced by the pontomedullary respiratory network after pontomesencephalic transection, the midbrain provides important modulation of respiration. Specifically, stimulation of the red nucleus elicits inspiratory inhibition, as manifest in the phrenic neurogram, in addition to excitation and inhibition of individual medullary respiratory-related units, with the majority of premotor units that receive rubral modulation being inhibited. Stimulation of the red nucleus also induces respiratory phase transitions, which appear to be pontine independent. These effects might be mediated by rubrobulbar and/or rubrospinal tracts. Although lesioning of the red nucleus does not alter respiration in normoxic conditions, it eliminates hypoxic ventilatory depression, which is the second phase of the biphasic ventilatory response to low oxygen tension. The finding that the red nucleus also plays a role in anti-nociception suggests that it might coordinate respiratory responses during noxious stimulation and, given that the red nucleus regulates upper limb flexors, it might represent one region in a distributed bulbar network contributing to respiratory-locomotor integration. Modulation of jaw opening by the red nucleus would support a model whereby it coordinates oromotor activity with breathing. Thus, the multiplicity of roles played by the red nucleus aptly position it to coordinate respiration in a variety of behavioural states. In this review, we seek to highlight the different features and regional specializations of the rubral contribution to respiratory control and underscore its vital importance to breathing in the freely behaving mammal.

4-Aminopyridine antagonizes saxitoxin- and tetrodotoxin-induced cardiorespiratory depression

Antagonism of saxitoxin- and tetrodotoxin-induced lethality by 4-aminopyridine was studied in urethane-anesthetized guinea pigs instrumented for the concurrent recordings of medullary respiratory-related unit activities (Bötzinger complex and Nu. para-Ambiguus), diaphragmatic electromyogram, electrocorticogram, Lead II electrocardiogram, blood pressure, end-tidal CO 2 and arterial O 2/CO 2/pH. The toxin (either saxitoxin or tetrodotoxin) was infused at a dose rate of 0.3 μg/kg/min (i.v.) to produce a state of progressive cardiorespiratory depression. The animals were artificially ventilated when the magnitude of integrated diaphragm activities was reduced to 50% of control. Immediately after the disappearance of the diaphragm electromyogram, the toxin infusion was terminated, and 4-aminopyridine (2 mg/kg, i.v.) was administered. The therapeutic effect of 4-aminopyridine was striking in that the toxin-induced blockade of diaphragmatic neurotransmission, vascular hypotension, myocardial anomalies, bradycardia and aberrant discharge patterns of medullary respiratory-related neurons could all be promptly restored to a level comparable to that of control condition. The animals were typically able to breathe spontaneously within minutes after 4-aminopyridine. At the dose level used to achieve the desired therapeutic responses, 4-aminopyridine produced no sign of seizure and convulsion. Although less serious side-effects such as cortical excitant/arousal and transient periods of fascicular twitch could be observed, these events were of minor concern, in our opinion, particularly in view of the remarkable therapeutic effects of 4-aminopyridine.

Central and peripheral cardio-respiratory effects of saxitoxin (STX) in urethane-anesthetized guinea-pigs

F.-C. T. Chang, B. J. Benton, R. A. Lenz and B. R. Capacio. Central and peripheral cardio-respiratory effects of saxitoxin (STX) in urethane-anesthetized guinea-pigs. Toxicon 31, 645–664, 1993.—Effects of saxitoxin (STX; 10 μg/kg; i.p.) on cardio-respiratory activites were evaluated in urethane-anesthetized guinea-pigs. Concurrent recordings were made of electrocorticogram (ECoG), bulbar respiratory-related unit activities, diaphragmatic electromyogram (DEMG), electrocardiogram (Lead II ECG), blood pressure, heart rate, end-tidal CO 2, arterial O 2/CO 2 tensions, and arterial pH. The average time to STX-induced respiratory failure was about 10 min. The most striking effect prior to apnea was a state of progressive bradypnea which emerged 5–7 min after the toxin administration. Other noteworthy responses included (i) a time-dependent decrease in ECoG amplitudes which typically began before the development of a bradypneic profile; (ii) an increasing degree of diaphragm neuromuscular blockade; (iii) a state of combined hypercapnia and uncompensated acidemia; (iv) a declining blood pressure; (v) an incrementally dysfunctional myocardial performance; and (vi) an increasingly degenerative central respiratory activity profile which ultimately culminated in a complete loss of central respiratory drive. The therapeutic effect of intratracheally administered oxygen was equivocal in that the cardio-respiratory activities, be they of central of peripheral nature, remained conspicuously dysfunctional and precarious despite 100% oxygen ventilation. What can be inferred from this study is two-fold. First, STX-induced ventilatory insufficiency can be attributed to a loss of functional integrity of both central and peripheral respiratory system components. That is, although diaphragm blockade contributes significantly to STX-induced respiratory failure, analyses of single respiratory unit activity data reveale d that the central respiratory rhythmogenic mechanism also appeared to play a pivotal role in the development of a bradypneic profile which promotes, and directly causes, a complete loss of respiratory drive. Second, a state of unabating depression of central respiratory activities, which seemed to be refractory to the effect of O 2, suggests STX has a direct and persistent action on medullary rhythmogenic mechanisms. In conclusion, these findings indicate that both central and peripheral cardio-respiratory components are critically involved in STX-induced apnea, dysfunctional cardiovascular performance, and lethality.

Modification of medullary respiratory-related discharge patterns by behaviors and states of arousal.

The modulatory influences of behaviors and states of arousal on bulbar respiratory-related unit (RRU) discharge patterns were studied in an unanesthetized, freely behaving guinea pig respiratory model system. When fully instrumented, this model system permits concurrent monitoring and recording of (i) single units from either Bötzinger complex or nucleus para-ambiguus; (ii) electrocorticogram; and, (iii) diaphragmatic EMG. In addition to being used in surveys of RRU discharge patterns in freely behaving states, the model system also offered a unique opportunity in investigating the effects of pentobarbital on RRU discharge patterns before, throughout the course of, and during recovery from anesthesia. In anesthetized preparations, a particular RRU discharge pattern (such as tonic, incrementing or decrementing) typically displayed little, if any notable variation. The most striking development following pentobarbital was a state of progressive bradypnea attributable to a significantly augmented RRU cycle duration, burst duration and an increase in the RRU spike frequencies during anesthesia. In freely behaving states, medullary RRU activities rarely adhered to a fixed, immutable discharge pattern. More specifically, the temporal organization (such as burst duration, cycle duration, and the extent of modulation of within-burst spike frequencies) of RRU discharge patterns regularly showed complex and striking variations, not only with states of arousal (sleep/wakefulness, anesthesia) but also with discrete alterations in electrocorticogram (ECoG) activities and a multitude of on-going behavioral repertoires such as volitional movement, postural modification, phonation, mastication, deglutition, sniffing/exploratory behavior, alerting/startle reflexes. Only during sleep, and on occasions when the animal assumed a motionless, resting posture, could burst patterns of relatively invariable periodicity and uniform temporal attributes be observed. RRU activities during sniffing reflex is worthy of further note in that, based on power spectrum analyses of concurrently recorded ECoG activities, this particular discharge pattern was clearly associated with the activation of a 6-10 Hz theta rhythm. These findings indicated that bulbar RRU activity patterns are subject to change by not only behaviors and sleep/wakefulness cycles, but also a variety of modulatory influences and feedback/feedforward biases from other central and peripheral physiological control mechanisms.

Antagonistic effects of somatostatin and substance P on respiratory regulation in the rat ventrolateral medulla oblongata

Substance P (SP) in the dose range 0.75–1.5 nmol exerts a potent stimulatory effect on ventilation after microinjection into the rat ventrolateral medulla oblongata (VLM; n. reticularis lateralis, n. paragigantocellularis lateralis). A significant but less pronounced effect is also seen in the dorsal medulla (DM; n. tractus solitarius). Somatostatin (0.6–1.8 nmol) inhibited ventilation and induced apnoea after microinjection into the VLM but not the DM. Serial microinjections of the two peptides showed a reciprocal antagonistic action in the VLM but not in the DM. The apnoea-inducing effect of SOM was blunted by SP while SOM reduced the ventilatory stimulation induced by SP. Extracellular single unit recordings were performed following the microiontophoretic application of SP and/or SOM to respiratory-related and non-respiratory-related neurons in the VLM and DM. Although a heterogeneous population of neurons were recorded from, the majority of respiratory-related units in the VLM responded with excitation to SP and inhibitory to SOM. A direct interaction between the peptides was seen in some respiratory-related units. The neurons not responding to either of the peptides were usually non-respiratory. Dorsal to the VLM, the type of response to the two peptides was less likely to be antagonistic and a wider distribution of response types were recorded. The results indicate a direct physiological antagonism between SP and SOM regarding their effects on respiratory regulation elicited in the VLM.

Effects of pentobarbital on respiratory functional dynamics in chronically instrumented guinea pigs

Respiratory effects of sodium pentobarbital (35 mg/kg; IP) were studied in guinea pigs chronically instrumented to permit concurrent recordings of bulbar respiratory-related units (RRUs), diaphragmatic electromyogram (DEMG), and electrocorticogram (ECoG). RRU activities were recorded from either the Bötzinger Complex (BOT; expiratory) or Nucleus para-Ambiguus (NpA; inspiratory). Pentobarbital-induced changes in respiratory-related activities were evaluated before, throughout the course of, and during recovery from, anesthesia. The most notable development following pentobarbital was a state of progressive bradypnea which was accompanied by a variety of complex changes in the amplitude and temporal attributes of RRU, DEMG and ECoG activities. As anesthetic effects progressed, the activity profiles of both BOT and NpA units underwent striking transformations from a behavioral and state-dependent wakefulness pattern to an activity profile characterized by i) a significantly augmented RRU cycle duration, burst duration and spike frequency; and, ii) an alteration to the pattern of within-burst spike frequency modulation. Along with changes in RRU activity, pentobarbital also produced a marked attenuation of the amplitudes of diaphragmatic activity as well as a discrete, time-dependent alteration in the amplitude and spectral characters of ECoG activities. Differences in BOT and NpA unit responses to alveolar CO 2 loading (ramp; 2% and 5%) across wakefulness and anesthesia states were also considerable. In addition to a depressed responsiveness to CO 2, the temporal attributes of BOT and NpA activity profiles also indicated an asymmetrical change under pentobarbital anesthesia. Taken together, these findings indicate that pentobarbital causes not only a fundamental alteration in bulbar rhythmogenic mechanisms, but also a differential influence on bulbar respiratory system components that are involved in the definition of the shape and the amplitude of central respiratory drive. In conclusion, this study offers, for the first time, direct evidence from physiologically and structurally intact preparations that the functional dynamics of respiratory system components are profoundly altered during pentobarbital anesthesia.

Respiratory and cardiovascular effects of tetrodotoxin in urethane-anesthetized guinea pigs

Cardiorespiratory effects of tetrodotoxin (TTX) (15 μg/kg, i.p.) were investigated in urethane-anesthetized guinea pigs acutely instrumented for the recording of medullary respiratory-related units (RRUs), diaphragm electromyogram (DEMG), electrocorticogram (ECoG), electrocardiogram (ECG), blood pressure (BP), endtidal CO 2, and arterial O 2 and CO 2. Respiratory system responses showed a hyperventilatory profile during the initial stage of intoxication. This was followed by an abrupt onset of a progressive decrease in the respiratory frequency, and a respiratory rate depression-related respiratory failure. The average time to TTX-induced respiratory arrest and death was 10.3 ± 4.2 min. Concurrentlyecorded inspiratory and expiratory RRU activities indicated that respiration invariably failed in an end-expiratory. position as manifested by a sustained period of expiratory RRU discharge. The progressive rate depression prior to respiratory arrest was temporally correlated only to a concomitantly augmenting expiratory RRU discharge duration. Inspiratory RRU discharge duration, on the other hand, did not display any significant change throughout the course of intoxication. The asymmetry in RRU response patterns indicates either an expiratory network component's particular sensitivity to perturbation by TTX or a dissociative trend in some bulbar respiratory rhythmogenic mechanisms. Peripheral cardiorespiratory changes were also quite profound. These included a gradual and steadfast decline in BP, a steadily decreasing amplitude in DEMG oscillations, and a state of progressive hypercapnia and hypoxenia. Changes in heart rate and ECG waveform attributes prior to respiratory arrest were not appreciable. In conclusion, in addition to a variety of TTX-induced peripheral cardiorespiratory effects, findings from this study have revealed a central respiratory system component that appears to show an unusual sensitivity to perturbation by TTX. The significance of this unique phenomenon as it relates to the nature and extent of TTX-induced central respiratory depression is discussed.

Neurophysiological concomitants of soman-induced respiratory depression in awake, behaving guinea pigs

Soman-induced respiratory failure was investigated in awake, behaving guinea pigs chronically instrumented to allow concurrent recordings of medullary respiratory-related unit (RRU) activity, diaphragm electromyogram (DEMG), and electrocorticogram. Responses to soman typically began with hyperpnea. Loss of consciousness, as indicated by the development of seizure activities, took place shortly after the onset of hyperpnea. This was followed by dyspnea, hypopnea, and, finally, respiratory failure. The most profound respiratory dysfunctions were seen during the development of dyspnea characterized by a progressively degenerative RRU-DEMG phase relationship (phase anomalies) and mixed patterns of ataxic breathing. Electrophysiographic records indicated that the anomalous RRU-DEMG phase phenomenon is attributable to a state of functional dissociation in some brainstem mechanisms that are normally involved in the orchestration of a synchronous respiratory drive. The failure of bulbar rhythmogenic mechanisms to maintain an orderly and synchronous recruitment of respiratory drive, which led to untimely and chaotic activations of respiratory muscles, was apparently the underlying cause of various ataxic breathing patterns and a reduced ventilatory efficiency. Spectral analyses of DEMG activities showed that, despite episodic muscle fasciculations and signs of fatigue, the functional integrity of the diaphragm was not significantly compromised by soman at a dose sufficient to produce respiratory failure. These findings not only support the notion of a relatively more important involvement of central respiratory mechanisms in soman-induced respiratory failure, but also identify a state of functional dissociation of central respiratory timing mechanisms as being a significant component in soman intoxication.

Control of discharge patterns of medullary respiratory neurons by pulmonary vagal afferent inputs.

To provide a better understanding of the central mechanisms by which pulmonary afferents reflexly control breathing, the responses of single respiratory neurons to vagal afferent patterns were analyzed. Respiratory-related unit (RRU) recordings were obtained from inspiratory (I), expiratory (E), and phase-spanning neurons in the ventral medulla of halothane-anesthetized, paralyzed, ventilated, vagotomized, mongrel dogs. Electrical stimulation of the largest vagal fibers was used to reflexly alter I and E durations (TI and TE) and to present various temporal input patterns to RRU. The net response was quantified by taking the difference between cycle-triggered histograms (CTH) of activity obtained during an input and the spontaneous control (no input) CTH. For step frequency patterns confined to either the I or E phase, 127 responses in 41 neurons were analyzed. The average step response time was greater than 500 ms. In general the time courses of the control and test-input discharge patterns were linearly related to one another. For I neurons the slopes (beta) of these relationships were linear functions of the vagal step frequency (Fv). Linear relationships were also obtained for 1/TI vs. Fv and 1/beta vs. TI. These results suggest that the vagal control of the discharge patterns of these neurons and phase timing is mediated via a process similar to gain modulation.

Firing patterns of bulbar respiratory neurones during sniffing in the conscious, non-paralyzed rabbit

On the basis of their firing patterns, 143 of 189 cells recorded in the respiratory centres of conscious, non-paralyzed rabbits could be identified as respiratory related units (RRUs). The activities of 45 RRUs were analyzed during sniffing in response to an odour stimulus. During sniffing as well as during normal breathing neuronal activity and respiratory movements were synchronized. Synchrony was lost, however, at the start of sniffing, indicating an action of the olfactory system on the respiratory generating mechanisms.

Phrenic afferent input to the lateral medullary reticular formation of the cat

The afferent inputs from phrenic nerve stimulation to the lateral reticular formation of the lower brain stem were studied in anesthetized spontaneously breathing cats. The activity of reticular neurons was recorded by means of extracellular tungsten microelectrodes. Electrical stimulation of the central end of the right phrenic nerve evoked excitatory or inhibitory responses in the lateral reticular nucleus (LRN), in the nucleus ambiguus (AMB) and in a region dorsal to the AMB of ipsi- and contralateral sides. Phrenic afferents belonging to the flexor reflex afferent group were involved in these responses. The discharge pattern of the respiratory related units (RRU) of the AMB were exceptionally affectec by phrenic nerve stimulations. It is concluded that high threshold phrenic relay in the LRN before projecting to the cerebellar cortex. The overlapping of respiratory and non-respiratory afferents in the reticular formation may participate to the adaptations of respiratory and somatomotor functions during specific behaviors.

Respiratory neurons of the pneumotaxic center during sleep and wakefulness

Extracellular recordings were made from respiratory related units (RRUs) in the pneumotaxic center (PNC) of unanesthetized cats across states of consciousness: wakefulness, NREM and REM sleep. Contrary to expectations from results in acute preparations, well-modulated respiratory activity was detected in chronic animals with vagi intact. Pontine RRU activity decreased during sleep in 75% of the cells studied. The greatest reduction occurred during REM sleep.

Quantitative study of anatomical distribution of respiration related neurons in the pons

The upper and middle pons and the lower mesencephalon of the cat were explored with extracellular microelectrodes in a search for respiratory related units (RRU). The preparation used, the unanesthetized normocapnic cat, resulted in an “isolated respiratory center” by eliminating input from, as well as output to, lung and thorax. Observations were done in standard experimental conditions which allowed quantitative comparisons of anatomical structures with respect to RRU density (RRUD), RRUD to total unit density (UD) ratio, and a respiratory modulation index (RMI).