Kathleen S. Henderson

Peripheral chemoreceptors determine the respiratory sensitivity of central chemoreceptors to CO2

We assessed the contribution of carotid body chemoreceptors to the ventilatory response to specific CNS hypercapnia in eight unanaesthetized, awake dogs. We denervated one carotid body (CB) and used extracorporeal blood perfusion of the reversibly isolated remaining CB to maintain normal CB blood gases (normoxic, normocapnic perfusate), to inhibit (hyperoxic, hypocapnic perfusate) or to stimulate (hypoxic, normocapnic perfusate) the CB chemoreflex, while the systemic circulation, and therefore the CNS and central chemoreceptors, were exposed consecutively to four progressive levels of systemic arterial hypercapnia via increased fractional inspired CO2 for 7 min at each level. Neither unilateral CB denervation nor CB perfusion, per se, affected breathing. Relative to CB control conditions (normoxic, normocapnic perfusion), we found that CB chemoreflex inhibition decreased the slope of the ventilatory response to CNS hypercapnia in all dogs to an average of 19% of control values (range 0–38%; n= 6), whereas CB chemoreflex stimulation increased the slope of the ventilatory response to CNS hypercapnia in all dogs to an average of 223% of control values (range 204–235%; n= 4). We conclude that the gain of the CNS CO2/H+ chemoreceptors in dogs is critically dependent on CB afferent activity and that CNS–CB interaction results in hyperadditive ventilatory responses to central hypercapnia.

Contribution of the carotid body chemoreceptors to eupneic ventilation in the intact, unanesthetized dog

We used extracorporeal perfusion of the reversibly isolated carotid sinus region to determine the effects of specific carotid body (CB) chemoreceptor inhibition on eupneic ventilation (Vi) in the resting, awake, intact dog. Four female spayed dogs were studied during wakefulness when CB was perfused with 1) normoxic, normocapnic blood; and 2) hyperoxic (>500 mmHg), hypocapnic ( approximately 20 mmHg) blood to maximally inhibit the CB tonic activity. We found that CB perfusion per se (normoxic-normocapnic) had no effect on Vi. CB inhibition caused marked reductions in Vi (-60%, range 49-80%) and inspiratory flow rate (-58%, range 44-87%) 24-41 s following the onset of CB perfusion. Thereafter, a partial compensatory response was observed, and a steady state in Vi was reached after 50-76 s following the onset of CB perfusion. This steady-state tidal volume-mediated hypoventilation ( approximately 31%) coincided with a significant reduction in mean diaphragm electromyogram (-24%) and increase in mean arterial pressure (+12 mmHg), which persisted for 7-25 min until CB perfusion was stopped, despite a substantial increase in CO(2) retention (+9 Torr, arterial Pco(2)) and systemic respiratory acidosis. We interpret these data to mean that CB chemoreceptors contribute more than one-half to the total eupneic drive to breathe in the normoxic, intact, awake animal. We speculate that this CB contribution consists of both the normal tonic sensory input from the CB chemoreceptors to medullary respiratory controllers, as well as a strong modulatory effect on central chemoreceptor responsiveness to CO(2).

The effects of locomotion on respiratory muscle activity in the awake dog

Using 6 chronically-instrumented awake dogs, we characterized the response of the respiratory muscles to mild and moderate treadmill exercise in terms of (1) the magnitude and timing of the EMGs of the costal and crural diaphragm, triangularis sterni and transverse abdominal muscles, and (2) the concomitant changes in esophageal (PE) and gastric (PG) pressures during treadmill exercise. Dogs wore a bivalved breathing mask which constrained breathing frequency and prevented some of the exercise-induced hypocapnia. Inspiratory and chest expiratory muscle EMGs increased linearly with a 1.5-fold tidal volume (VT) change during exercise. Abdominal muscle recruitment occurred during most exercise levels and exhibited (1) phasic activity coincident with footplant and (2) tonic activation evidenced by a baseline shift in PG and EMG activities. During control, inspiratory and expiratory flow preceded the onset of respiratory muscle EMG activity, but during exercise, electrical activation of these muscles coincided with the onset of mechanical flow. We conclude that phasic and tonic activation of expiratory muscles during exercise in the dog is significantly influenced by both respiratory and locomotory requirements. These patterns of expiratory muscle recruitment may assist inspiration by lengthening inspiratory muscles through decrements in end-expiratory lung volume and by passive recoil of the rib cage and abdominal compartments.

Respiratory muscle recruitment during selective central and peripheral chemoreceptor stimulation in awake dogs.

1. In four awake dogs we measured EMG activity of three inspiratory and four expiratory muscles during sustained central chemoreceptor stimulation (CO2 inhalation), and peripheral chemoreceptor stimulation (intravenous infusion of almitrine bismesylate (almitrine)). By using this selective pharmacological stimulation of the peripheral chemoreceptors and reversibly cold‐blocking pulmonary stretch receptors, we were able to determine the effects of each type of stimulation on respiratory muscle recruitment in the absence of such complicating influences as pulmonary stretch receptor feedback, cerebral hypoxia or hypocapnia, and differences in breathing pattern. 2. During 10 min of steady‐state hyperpnoea (minute ventilation VI, approximately twice eupnoea) caused by either hypercapnia or isocapnic stimulation of the carotid bodies with almitrine, all three inspiratory and all four expiratory muscles demonstrated significant and sustained elevations in EMG activity. 3. With both types of chemoreceptor stimulation, as tidal volume, VT, increased, so did the mean electrical activities of the crural diaphragm (r = 0.88), costal diaphragm (r = 0.93), parasternals (r = 0.82), triangularis sterni (r = 0.74), transversus abdominis (r = 0.77), external obliques (r = 0.68) and internal intercostals (r = 0.75). 4. In each dog, the response of ventilation and of the diaphragmatic EMG to a given level of central or peripheral chemoreceptor stimulation is highly reproducible from one test day to the next. On the other hand, accessory inspiratory and expiratory abdominal and rib cage muscles in two of the four dogs showed highly significant changes from day to day in the amount of their EMG activity at any given VT. 5. During steady‐state ventilatory stimulation, 2 min intervals were chosen during which the two types of chemoreceptor stimulation had caused hyperpnoeas with similar values for VT, total time per breath (TTOT) and inspiratory time divided by the total time (TI/TTOT). Comparison of EMG activities during these matched hyperpnoeas revealed that there were no differences in the activities of any of the muscles between the two forms of stimulation. We conclude that peripheral chemoreceptor stimulation causes significant and sustained recruitment of expiratory muscles even in the absence of pulmonary feedback and that both expiratory and inspiratory muscles are recruited to the same extent during peripheral chemoreceptor stimulation as they are during an identical hyperpnoea caused by central chemoreceptor stimulation.

Peripheral chemoreceptors determine the respiratory sensitivity of central chemoreceptors to CO2: role of carotid body CO2

The influence of specific carotid body (CB) normoxic hypocapnia, hypercapnia and normocapnia on the ventilatory sensitivity of central chemoreceptors to systemic hypercapnia was assessed in seven awake dogs with extracorporeal perfusion of the vascularly isolated CB. Chemosensitivity in this preparation was similar to that in the intact animal. Separation of CB circulation from that of the brain was confirmed. When the isolated CB was hypercapnic vs. hypocapnic and when the isolated CB was normocapnic vs. hypocapnic, the group mean central CO2 response slopes of minute ventilation ( V̇I ) (P ≤ 0.01) and mean inspiratory flow rate (VT/TI) (P ≤ 0.05) increased significantly. Tidal volume (VT), breathing frequency (fb)and rate of rise of diaphragm EMG were increased in 6 of 7 dogs but did not achieve statistical significance. We propose that hyperaddition is the dominant form of chemoreceptor interaction under conditions of quiet wakefulness in intact animals and over a wide range of CB PCO2 and PO2 .

Pulmonary-locomotory interactions in exercising dogs and horses

In exercising quadrupeds, limb movement is often coupled with breathing frequency. This finding has lead some investigators to conclude that locomotory forces, associated with foot plant, abdominal visceral displacements or lumbo-sacral flexion, are the primary determinants of airflow generation. Analysis of respiratory muscle electrical activation (EMG) and contraction profiles in chronically instrumented dogs and horses, along with measurements of esophageal pressure (Pes) changes and limb movements, provide evidence that each breath during the exercise hyperpnea is determined by respiratory neuromuscular events. Specifically: (1) Phasic diaphragmatic EMG and tidal shortening are always synchronous with decreases in Pes; (2) decrements in Pes are always associated with inspiratory flow generation; and (3) strict phase coupling between breathing and stride frequency is not obligatory. Thus, although locomotory-associated forces may minimally assist with flow generation, they are not the primary determinants of breathing during exercise.

The effects of chemical versus locomotory stimuli on respiratory muscle activity in the awake dog

Using 6 chronically instrumented awake dogs, we contrasted the ventilatory and respiratory muscle EMG responses to steady-state normoxic hypercapnia with those occurring at similar levels of ventilation during steady-state treadmill exercise. During hypercapnia, increases in ventilatory output were primarily due to increments in VT whereas during exercise, increases in minute ventilation were due primarily to increases in frequency. With either hyperpnea, augmentation of both inspiratory and expiratory muscle EMG activity occurred but only diaphragmatic EMG activity was strongly correlated with tidal volume changes in both conditions. Using both EMGs and thoraco-abdominal pressures, we found that with increasing chemical or locomotory stimuli, during inspiration (1) diaphragmatic EMG activation occurred sooner relative to the onset of mechanical flow and (2) that respiratory muscles other than the diaphragm contributed significantly to the generation of inspiratory flow and VT either actively or passively through recoil. Post-inspiratory inspiratory activity of the crural diaphragm shortened relative to control in hypercapnia but did not change in exercise. Finally, during mild hyperpneas, end-expiratory lung volume did not change significantly in either hypercapnia or exercise but decreased at the higher levels of hyperpnea only during exercise. This may reflect differences not in the magnitude of phasic expiratory muscle activity, but rather in the degree of tonic abdominal muscle activation between the two conditions.

The influence of carotid body chemoreceptors on expiratory muscle activity

Respiratory muscle EMG responses to transient, specific carotid body excitation (NaCN, hypoxia) and inhibition (dopamine, hyperoxia) were studied in 7 unanesthetized standing dogs while intact (N = 7) and during bilateral cold block of the vagi (N = 3). In all dogs carotid body excitation augmented the EMG activities of both expiratory muscles (triangularis sterni, TS; transversus abdominis, TA) and the (inspiratory) crural diaphragm (CR); carotid body inhibition significantly reduced phasic EMG activities in all muscles. With vagal blockade the response of the TS to carotid body stimulation was increased; that of the TA was essentially unchanged from the intact state. In all dogs expiratory muscle recruitment in response to carotid body excitation/inhibition could either precede or follow that of the CR. We conclude that in response to specific, transient carotid body excitation or inhibition: (1) Changes in CR, TS and TA EMGs are qualitatively similar. (2) Vagal feedback is strongly inhibitory to TS and excitatory to TA. (3) Changes in expiratory muscle EMGs do not require preceding changes in the CR. Carotid body stimulation contributes significantly to the generation of phasic expiratory activity during eupnea.

Ventilatory instability induced by selective carotid body inhibition in the sleeping dog

There is a considerable evidence that hypocapnia is a major contributor to the genesis of central apnea and periodic breathing (PB) in humans during sleep. Studies using mechanical ventilation to lower the arterial carbon dioxide pressure (PaCO2) have shown that during non-rapid eye movement sleep (NREM), when respiration is under predominantly metabolic control, there is a highly sensitive apneic threshold (AT) induced by reduction in PaCO2 that were only 2 to 4 mm Hg less than eupneic PaCO2.1, 2, 3, 4 If there is a small difference between the AT for CO2 (PATCO2) and the eupneic PaCO2 (a narrow “CO2 reserve”), then a relatively small increase in ventilation (ventilatory overshoot), from whatever cause, could result in apnea. Conversely, if there is a large difference between eupneic PaCO2 and PATCO2 (a wide “CO2 reserve”) then a larger increment of ventilation is required to produce apnea.

Interactive Ventilatory Effects of Carotid Body Hypoxia and Hypocapnia in the Unanesthetized Dog

In a previous study (5) we demonstrated that moderate normoxic hypocapnia, when limited to the carotid sinus region (carotid body), could markedly inhibit ventilation and VT in the awake or sleeping (Non-REM and REM) dog. TI and TE were not changed significantly, thus there was no tendency for carotid body hypocapnia to produce apnoeas. The ventilatory inhibition was graded over the range of carotid body hypocapnia (ΔPCBCO2 = -7 to -15 Torr relative to eupnoea) but the stimulus: response slope was most sensitive over the eupnoea to -7 Torr range at least during wakefulness and Non-REM sleep. In other words, in these conditions there was clearly no “dog-leg” present in the ventilatory response to carotid body hypocapnia (2). During REM sleep there was no consistent ventilatory response until ΔPCBCO2 was > -7 Torr (Fig. 1). During wakefulness and Non-REM sleep the inhibitory effect was probably maximal between about -10 and -15 Torr as perfusion of the carotid body with normocapnic and hyperoxic blood (PCBO2>500 Torr) produced ventilatory inhibition that was essentially identical in time course, magnitude and ventilatory pattern to that produced by -10 to -15 Torr carotid body hypocapnia. Given the well known inhibitory effects of hyperoxia on carotid sinus nerve output (3,4) we concluded that carotid body hypocapnia in the -10 to -15 range must virtually silence chemoreceptor output in the carotid body.