Bruce G. Lindsey

Functional connectivity among ventrolateral medullary respiratory neurones and responses during fictive cough in the cat

1 This study tested predictions from a network model of ventrolateral medullary respiratory neurone interactions for the generation of the cough motor pattern observed in inspiratory and expiratory pump muscles. 2 Data were from 34 mid‐collicularly decerebrated, paralysed, artificially ventilated cats. Cough‐like patterns (fictive cough) in efferent phrenic and lumbar nerve activities were elicited by mechanical stimulation of the intrathoracic trachea. Neurones in the ventral respiratory group, including the Bötzinger and pre‐Bötzinger complexes, were monitored simultaneously with microelectrode arrays. Spike trains were analysed for evidence of functional connectivity and responses during fictive cough with cycle‐triggered histograms, autocorrelograms, cross‐correlograms, and spike‐triggered averages of phrenic and recurrent laryngeal nerve activities. 3 Significant cross‐correlogram features were detected in 151 of 1988 pairs of respiratory modulated neurones. There were 59 central peaks, 5 central troughs, 11 offset peaks and 2 offset troughs among inspiratory neurone pairs. Among expiratory neurones there were 23 central peaks, 8 offset peaks and 4 offset troughs. Correlations between inspiratory and expiratory neurones included 20 central peaks, 10 central troughs and 9 offset troughs. Spike‐triggered averages of phrenic motoneurone activity had 51 offset peaks and 5 offset troughs. 4 The concurrent responses and multiple short time scale correlations support parallel and serial network interactions proposed in our model for the generation of the cough motor pattern in the respiratory pump muscles. Inferred associations included the following. (a) Excitation of augmenting inspiratory (I‐Aug) neurones and phrenic motoneurones by I‐Aug neurones. (b) Inhibition of augmenting expiratory (E‐Aug) neurones by decrementing inspiratory (I‐Dec) neurones. (c) Inhibition of I‐Aug, I‐Dec and E‐Aug neurones by E‐Dec neurones. (d) Inhibition of I‐Aug and I‐Dec neurones and phrenic motoneurones by E‐Aug neurones. The data also confirm previous results and support hypotheses in current network models for the generation of the eupnoeic pattern.

Medullary respiratory neurones and control of laryngeal motoneurones during fictive eupnoea and cough in the cat

1 This study addressed the hypothesis that ventrolateral medullary respiratory neurones participate in the control of laryngeal motoneurones during both eupnoea and coughing. 2 Data were obtained from 28 mid‐collicular decerebrated, artificially ventilated cats. Cough‐like motor patterns (fictive cough) in phrenic, lumbar and recurrent laryngeal nerves were elicited by mechanical stimulation of the intrathoracic trachea. Microelectrode arrays were used to monitor simultaneously several neurones in the ventral respiratory group, including the Bötzinger and pre‐Bötzinger complexes. Spike trains were evaluated for responses during fictive cough and evidence of functional connectivity with spike‐triggered averages of efferent recurrent laryngeal nerve activity. 3 Primary features were observed in averages triggered by 94 of 332 (28 %) neurones. An offset biphasic wave with a positive time lag was present in the unrectified average for 10 inspiratory and 13 expiratory neurones. These trigger neurones were respectively identified as inspiratory laryngeal motoneurones with augmenting, decrementing, plateau and ‘other’ discharge patterns, and expiratory laryngeal motoneurones with decrementing firing patterns. 4 Rectified averages triggered by inspiratory neurones included 37 offset peaks, 11 central peaks and one offset trough. Averages triggered by expiratory neurones had 12 offset peaks, six central peaks and four offset troughs. Relationships inferred from these features included premotor actions of inspiratory neurones with augmenting, decrementing, plateau and ‘other’ patterns on inspiratory laryngeal motoneurones, and premotor actions of decrementing and ‘other’ expiratory neurones on expiratory laryngeal motoneurones. Corresponding changes in neuronal firing patterns during fictive cough supported these inferences. 5 The data confirm and extend previous results on the control of laryngeal motoneurones during eupnoea and support the hypothesis that the same premotor neurones help to shape motoneurone firing patterns during both eupnoea and coughing.

Simulations of a ventrolateral medullary neural network for respiratory rhythmogenesis inferred from spike train cross-correlation

Connections among ventrolateral medullary respiratory neurons inferred from spike train analysis were incorporated into a model and simulated with the program SYSTM11 (MacGregor 1987). Inspiratory (I) and expiratory (E) neurons with augmenting (AUG) and decrementing (DEC) discharge patterns and rostral I-E/I neurons exhibited varying degrees of adaptation, but no endogenous bursting properties. Simulation parameters were adjusted so that respiratory phase durations, neuronal discharge patterns, and short-time scale correlations were similar to corresponding measurements from anesthetized, vagotomized, adult cats. Rhythmogenesis persisted when the strength of each set of connections was increased 100% over a smaller effective value. Changes in phase durations and discharge patterns caused by manipulation of connection strengths or population activity led to several predictions. (a) Excitation of the I-E/I population prolongs the inspiratory phase. (b) Rhythmic activity can be reestablished in the absence of I-E/I activity by unpatterned excitation of I-DEC and I-AUG neurons. (c) An increase in I-DEC neuron activity can cause an apneustic respiratory pattern. (d) A decrease in I-DEC neuron activity increases the slope of the inspiratory ramp and shortens inspiration, (e) Excitation of the E-DEC population prolongs the expiratory phase or produces apnea; inhibition of E-DEC neurons reduces expiratory time. (f) Excitation of E-AUG cells causes I-AUG neurons to exhibit a step rather than a ramp increase in firing rate at the onset of their active phase. The results suggest mechanisms by which the duration of each phase of breathing and neuronal discharge patterns may be regulated.

Long-term facilitation of phrenic nerve activity in cats: responses and short time scale correlations of medullary neurones.

1. Stimulation of either peripheral chemoreceptors or nucleus raphe obscurus results in long‐term facilitation of phrenic motoneurone activity. The first objective of this work was to measure the concurrent responses of neurones in the nucleus raphe obscurus, the nucleus tractus solitarii, and the regions of the retrofacial nucleus, nucleus ambiguus and nucleus retroambigualis during induction of long‐term facilitation. A second goal was to assess functional relationships of the chemoresponsive raphe neurones with neurones in the other monitored locations and with phrenic motoneurones. 2. Up to thirty single medullary neurones and phrenic nerve efferent activity were recorded simultaneously in fifteen anaesthetized, paralysed, vagotomized, artificially ventilated adult cats. Carotid chemoreceptors were stimulated by close arterial injection of 200 microliters of CO2‐saturated saline solution. Spike trains were analysed with cycle‐triggered histograms and two statistical tests for respiratory modulation. Peristimulus‐time histograms and cumulative sum histograms were used to assess responses to stimulation. Cross‐correlation was used to test for non‐random temporal relationships between spike trains. Spike‐triggered average histograms provided evidence for functional associations with phrenic motoneurones. 3. One hundred and thirteen of 348 neurones were monitored in the nucleus raphe obscurus. The firing rates of twenty‐nine raphe neurones increased during stimulation; eighteen decreased. In twenty‐one pairs of concurrently monitored raphe neurones, the firing rate of one increased its activity during stimulation then decreased, while the other showed an increase that began as the rate of the former declined. Eighteen chemoresponsive raphe neurones had short time scale features in their phrenic spike‐triggered averages. Short time scale features were found in cross‐correlograms from 184 of 1407 neurone pairs. 4. The data suggest parallel routes by which carotid chemoreceptors influence medullary raphe neurones and support the hypotheses that mid‐line respiratory‐related neuronal assemblies transform information from those receptors and regulate the gain of respiratory motor output.

MEDULLARY RAPHE NEURONES AND BARORECEPTOR MODULATION OF THE RESPIRATORY MOTOR PATTERN IN THE CAT

1 Perturbations of arterial blood pressure change medullary raphe neurone activity and the respiratory motor pattern. This study sought evidence for actions of baroresponsive raphe neurones on the medullary respiratory network. 2 Blood pressure was perturbed by intravenous injection of an α1‐adrenergic receptor agonist, unilateral pressure changes in the carotid sinus, or occlusion of the descending aorta in thirty‐six Dial‐urethane‐anaesthetized, vagotomized, paralysed, artificially ventilated cats. Neurones were monitored with microelectrode arrays in two or three of the following domains: nucleus raphe obscurus‐nucleus raphe pallidus, nucleus raphe magnus, and rostral and caudal ventrolateral medulla. Data were analysed with cycle‐triggered histograms, peristimulus time and cumulative sum histograms, cross‐correlograms and spike‐triggered averages of efferent phrenic nerve activity. 3 Prolongation of the expiratory phase and decreased peak integrated phrenic amplitude were most frequently observed. Of 707 neurones studied, 310 had altered firing rates during stimulation; changes in opposite directions were monitored simultaneously in fifty‐six of eighty‐seven data sets with at least two baroresponsive neurones. 4 Short time scale correlations were detected between neurones in 347 of 3388 pairs. Seventeen pairs of baroresponsive raphe neurones exhibited significant offset correlogram features indicative of paucisynaptic interactions. In correlated raphe‐ventrolateral medullary neurone pairs with at least one baroresponsive neurone, six of seven ventrolateral medullary decrementing expiratory (E‐Decr) neurones increased their firing rate during baroreceptor stimulation. Thirteen of fifteen ventrolateral medullary inspiratory neurones correlated with raphe cells decreased their firing rate during baroreceptor stimulation. 5 The results support the hypothesis that raphe neuronal assemblies transform and transmit information from baroreceptors to neurones in the ventral respiratory group. The inferred actions both limit and promote responses to sensory perturbations and match predictions from simulations of the respiratory network.

Changes in cat medullary neurone firing rates and synchrony following induction of respiratory long-term facilitation

1 Long‐term facilitation is a respiratory memory expressed as an increase in motor output lasting more than an hour. This change is induced by repeated hypoxia, stimulation of carotid chemoreceptors, or electrical stimulation of the carotid sinus nerve or brainstem mid‐line. The present work addressed the hypothesis that persistent changes in medullary respiratory neural networks contribute to long‐term facilitation. 2 Carotid chemoreceptors were stimulated by close arterial injection of CO2‐saturated saline solution. Phrenic nerve efferent activity and up to 30 single medullary neurones were recorded simultaneously in nucleus tractus solitarii (NTS) including the dorsal respiratory group (DRG), Bötzinger‐ventral respiratory group (Böt‐VRG), and nucleus raphe obscurus of nine adult cats, anaesthetized, injected with a neuromuscular blocking agent, vagotomized and artificially ventilated. 3 The firing rates of 87 of 105 neurones (83 %) changed following induction of long‐term facilitation. Nine of eleven DRG and Böt‐VRG putative premotor inspiratory neurones had increased firing rates with long‐term facilitation. Fourteen of twenty‐one raphe obscurus neurones with control firing rates less than 4 Hz had significant long‐term increases in activity. 4 Cross‐correlogram analysis suggested that there were changes in effective connectivity of neuron pairs with long‐term facilitation. Joint peristimulus time histograms and pattern detection methods used with ‘gravity’ analysis also detected changes in short time scale correlations associated with long‐term facilitation. 5 The results suggest that changes in firing rates and synchrony of VRG and DRG premotor neurones and altered effective connectivity among other functionally antecedent elements of the medullary respiratory network contribute to the expression of long‐term facilitation.

Inspiratory drive and phase duration during carotid chemoreceptor stimulation in the cat: medullary neurone correlations.

1. This study addressed the hypothesis that there is a parallel processing of input from carotid chemoreceptors to brainstem neurones involved in inspiratory phase timing and control of inspiratory motor output amplitude. Data were from fifteen anaesthetized, bilaterally vagotomized, paralysed, artificially ventilated cats. Carotid chemoreceptors were stimulated by close arterial injection of 200 microliters of CO2‐saturated saline solution. 2. Planar arrays of tungsten microelectrodes were used to monitor simultaneously up to twenty‐two neurones in the nucleus tractus solitarii (NTS) and ventral respiratory group (VRG). Spike trains were analysed with two statistical tests of respiratory modulation, cycle‐triggered histograms, peristimulus‐time histograms, cumulative sum histograms and cross‐correlograms. 3. In NTS, 16 of 26 neurones with respiratory and 12 of 27 without respiratory modulation changed firing rate during carotid chemoreceptor stimulation. In the VRG 72 of 112 respiratory and 14 of 48 non‐respiratory neurones changed firing rate during stimulation. 4. The spike trains of 85 of 1276 pairs (6.7%) of cells exhibited short time scale correlations indicative of paucisynaptic interactions. Ten pairs of neurones were each composed of a rostral VRG phasic inspiratory neurone that responded to carotid chemoreceptor stimulation with a decline in firing rate and a caudal VRG phasic inspiratory neurone that increased its firing rate. Cross‐correlograms from two of the pairs had features consistent with excitation of the caudal neurones by the rostral cells. A decrease in the duration of activity of the rostral VRG neurones was paralleled by the decrease in inspiratory time of phrenic nerve activity. Caudal VRG inspiratory neurones increased their activity as phrenic amplitude increased. Spike‐triggered averages of all four neurones indicated post‐spike facilitation of phrenic motoneurones. 5. The results support the hypothesis that unilateral stimulation of carotid chemoreceptors results in parallel actions. (a) Inhibition of rostral VRG I‐Driver neurones decreases inspiratory duration. (b) Concurrent excitation of premotor VRG and dorsal respiratory group inspiratory neurones increases inspiratory drive to phrenic motoneurones. Other data suggest that responsive ipsilateral neurones act to regulate contralateral neurones.

Cardio-respiratory coupling depends on the pons

Cardio-respiratory coupling is reciprocal; it is expressed as respiratory-modulated sympathetic nerve activity and pulse-modulated respiratory motor activity. In the brainstem, the neuraxis controlling cardio-respiratory functions forms a ventrolateral cell column which extends to the dorsolateral (dl) pons. Our general working hypothesis is that these control systems converge at points with the common purpose of gas exchange and that neural activity along this axis coordinates both arterial pulse pressure and breathing. Here, we review the data showing that pontine nuclei modulate heart rate, blood pressure and breathing, and present new results demonstrating a vagal influence on pontine activity modulated with both arterial pulse pressure and phrenic nerve activity in the decerebrate cat. Generally with the vagi intact, dl pontine activity was weakly modulated by both arterial pulse pressure and respiratory pattern. After bilateral vagotomy, the strength and consistency of respiratory modulation increased significantly, although the strength and consistency of arterial pulse pressure modulation did not change significantly for the group; a decrease in some (62%) was offset by an increase in others (36%) neurons. Thus, the vagus shapes the envelope of the cycle-triggered averages of neural activity for both the respiratory and cardiac cycles. These data provide insight into the neural substrate for the prominent vagal effect on the cardio-respiratory coupling pattern, in particular respiratory sinus arrhythmia. While these results support convergence of inputs to neural populations controlling breathing and cardiovascular functions, the physiologic role of balancing ventilation, vascular resistance, heart rate and blood flow for the benefit of tissue oxygenation, remains hypothetical.

Respiratory neural activity during long-term facilitation

Intermittent hypoxia results in a long-term facilitation (LTF) of respiratory efferent activity. The studies reviewed here presented data from both anesthetized and decerebrate, paralyzed, vagotomized, artificially ventilated adult cats. Multiple arrays of tungsten microelectrodes were used to record the concurrent responses of brain stem neurons that contribute to respiratory motor pattern generation. Spike trains were analyzed with firing rate histograms, peristimulus time histograms, cycle triggered histograms, spike triggered averages with multiunit phrenic efferent activity, cross correlation histograms, joint peristimulus time histograms and the gravity method. These studies addressed several hypotheses. (1) There is parallel processing of input from carotid chemoreceptors to the brain stem. (2) Respiratory related midline neurons are involved in the induction and maintenance of LTF. (3) There is a change in effective connectivity of brain stem neurons with LTF. (4) Neural networks involved in the induction and maintenance of LTF have patterns of synchrony that recur with a frequency greater than expected by chance.

Coordination of cough and swallow: a meta-behavioral response to aspiration.

Airway protections is the prevention and/or removal of material by behaviors such as cough and swallow. We hypothesized these behaviors are coordinated to respond to aspiration. Anesthetized animals were challenged with simulated aspiration that induced both coughing and swallowing. Electromyograms of upper airway and respiratory muscles together with esophageal pressure were recorded to identify and evaluate cough and swallow. During simulated aspiration, both cough and swallow intensity increased and swallow duration decreased consistent with rapid pharyngeal clearance. Phase restriction between cough and swallow was observed; swallow was restricted to the E2 phase of cough. These results support three main conclusions: 1) the cough and swallow pattern generators are tightly coordinated so as to generate a protective meta-behavior; 2) the trachea provides feedback on swallow quality, informing the brainstem about aspiration incidences; and 3) the larynx and upper esophageal sphincter act as two separate valves controlling the direction of positive and negative pressures from the upper airway into the thorax.