James P. Kiley

Stimulation by central command of locomotion, respiration and circulation during exercise

We studied the relationships between exercise (locomotion) and respiratory and circulatory responses in 19 cats that walked or ran normally on a treadmill, and in 16 paralyzed animals during fictive locomotion, i.e., locomotory activity in motor nerves to the legs. Preparations included anesthetized cats with intact brains and unanesthetized decorticate (hypothalamic) and decerebrate (mesencephalic) animals. Spontaneous actual locomotion and fictive locomotion occurred in all preparations except the mesencephalic cats. Electrical stimulation or injection of a GABA antagonist (picrotoxin) into the hypothalamic locomotor region caused locomotion to develop. In all cases when locomotion occurred, respiration and arterial pressure increased in proportion to the level of locomotor activity despite control or ablation of respiratory feedback mechanisms. Respiration and arterial pressure increased similarly during fictive locomotion despite the absence of muscular contraction or limb movement and the lack of change of metabolic rate. We conclude that the study provides experimental support for the feed-forward, or command signal, hypothesis for the genesis of proportional changes of respiration and circulation that occur during exercise. Feedback mechanisms are not required for its operation. We suggest that command signals emanating from the hypothalamus provide the primary drive for changes of respiration and circulation during exercise.

Mechanism of respiratory effects of methylxanthines

Neural respiratory responses to theophylline, aminophylline and ethylenediamine were determined in paralyzed, vagotomized and glomectomized cats whose end-tidal PCO2 and brain temperature were kept constant. Intravenous theophylline and aminophylline similarly stimulated respiration, but ethylenediamine had no effect. The following did not cause the response: muscular and mechanical factors, carotid body and vagal reflexes, spinally mediated mechanisms arising below C7, changes of arterial PCO2 or medullary ECF pH, changes of whole body metabolic rate or release of substances from the adrenal glands. Absence of suprapontine brain did not prevent the response. Pretreatment with a serotonin antagonist did not affect the response but two different dopamine antagonists caused its attenuation. When administered into the third ventricle, theophylline did not stimulate respiration, but both aminophylline and ethylenediamine, due to the latters ability to mimic the inhibitory effects on neurons of gamma-aminobutyric acid (GABA), caused significant depression of respiration. We conclude that the neural respiratory response to systemically administered theophylline is mediated at the level of the brainstem, and somehow involves the action of the neurochemical dopamine. The failure of cerebroventricularly administered theophylline to stimulate respiration must be related to its inability to reach the appropriate neurons from the cerebrospinal fluid.

Respiratory effects of a long-acting analog of adenosine

The effect on respiration, measured as integrated phrenic nerve activity, of an analog of adenosine, N6(L-2-phenylisopropyl)adenosine (PIA), was determined in 26 paralyzed, vagotomized and glomectomized cats whose end-tidal pCO2 was kept constant by means of a servocontrolled ventilator. Whether given intravenously or into the third cerebral ventricle, PIA caused a dose-related depression of respiration, involving both tidal activity and respiratory frequency. In a group of 6 cats, the effects of intraventricular PIA or its vehicle alone on medullary extracellular fluid (ECF) pH were also determined. Vehicle alone had no effect on either ECF pH or respiratory activity. PIA was associated with the development over 10-20 min of a metabolic acidosis in the medulla, but still led to marked depression of respiration, thus ruling out an increase of medullary blood flow as cause of the decreased respiration. We conclude that the adenosine analog, PIA, acts to inhibit directly neurons in the brain that are involved in the control of respiration, and we suggest that adenosine may act as a tonic modulator of respiration.

Oscillations of medullary extracellular fluid pH caused by breathing

The purpose of this study was to determine if respiratory oscillations of arterial PCO2 are transmitted to the medullary ECF. Anesthetized, paralyzed cats whose vagi and carotid sinus nerves had been cut were studied. A flat surfaced pH electrode (2 mm in diameter) and a specially built differential amplifier were used to measure pH on the ventral surface of the medulla. Elimination of noise and high sensitivity were obtained by integrating the output from the pH amplifier with a digital voltmeter . We measured ECF pH oscillations that had the same period as the ventilator. The amplitude of the oscillations was about 0.006 unit when the ventilator was slow (less than 10/min). The amplitude decreased progressively as the rate was increased and oscillations were usually undetectable at rates above 25/min. Experiments were performed that ruled out mechanical artifact and ventilator-related fluctuations in arterial pressure as being causal. We conclude that oscillations of alveolar CO2 are transmitted to the brain by the circulation and result in oscillations of ECF pH.

The roles of medullary extracellular and cerebrospinal fluid pH in control of respiration

To determine the effective stimulus to the central chemoreceptors, we measured CSF and medullary extracellular fluid (ECF) pH and phrenic activity in 11 anesthetized, paralyzed, vagotomized and glomectomized cats. Flat-tipped pH electrodes (2 mm diam.) were used to measure ECF pH on the ventral surface of the medulla and CSF pH 2 mm above the surface. Changes in alveolar/arterial PCO2 were produced by airway occlusions of 10-20 sec durations. Changes in CSF PCO2 and pH were made by infusing 100% CO2 or an acid buffer into the CSF. Airway occlusion caused an increase of alveolar/arterial PCO2. ECF pH began to fall 6-10 sec later, with a maximum decrease of 0.032 pH unit at 21.9 sec. Phrenic activity increased as ECF pH decreased, the greatest activity occurring when ECF pH was most acid. CSF pH decreased after a longer delay. Its maximum decrease at 54.1 sec was smaller (0.026 pH unit) than ECF pH and did not correlate with the increase of phrenic activity. Addition of 100% CO2 or an acid buffer into the CSF produced an acid shift in the CSF pH but no change in ECF pH or phrenic activity. Prolonged (greater than 30 min) increase of acidity of CSF did not alter phrenic activity until ECF pH developed a delayed acid shift. Even then, the change of ECF pH was much smaller than that of CSF. We conclude that medullary chemoreceptors do not respond to changes of CSF pH or PCO2 and that change of pH of CSF minimally affects ECF pH. On the other hand, respiratory responses are closely linked to changes in ECF pH.

Effects of hyperoxia on medullary ECF pH and respiration in chemodenervated cats.

The effects of transitions from air-breathing to hyperoxia (100% O2), and the reverse, on respiration (phrenic activity) and on medullary extracellular fluid (ECF) pH, or hydrogen ion concentration [H+], were studied in 8 anesthetized, paralyzed, vagotomized and glomectomized cats whose end-tidal PCO2 was kept constant. The transition from air to hyperoxia (7 cats) led to a small (1.23 nmol/L [H+], 0.010 pH unit) acidic shift of medullary ECF and a 24% increase of neural tidal and minute respiratory activity with no significant change of frequency. Opposite changes of approximately equal magnitude followed the transition from hyperoxia to air (8 cats). We show that the slopes of the respiratory responses to changing ECF [H+] in the present study are not different than the slopes with CO2-induced changes of ECF [H+] in 25 other cats. Our findings indicate that most, if not all, of the respiratory increase after hyperoxia is due to accumulation of CO2 and H+ in medullary ECF that act on the central chemoreceptors. We suggest that decreased medullary blood flow and the Haldane effect are the main mechanisms causing the rise of medullary PCO2 and stimulation of breathing.

Brain, blood and rectal temperature during whole body cooling.

To determine if rectal temperature is an adequate index of brain temperature during changing thermal conditions, we measured rectal, cerebral cortical, and carotid arterial blood temperatures simultaneously during whole body cooling in adult cats. The mean steady state rectal, brain and carotid arterial temperatures at the onset of cooling were: 39.2 +/- 0.2, 38.5 +/- 0.2, and 38.3 +/- 0.3 degrees C, respectively. Rectal temperature decreased faster than both brain and arterial blood, while only a small temperature difference was observed between brain and arterial blood, brain always exceeding blood. Rectal temperature cannot be considered an adequate index of brain temperature. Carotid arterial temperature is a better estimate of brain temperature.

Effect of graded cooling of intermediate areas on respiratory response to vagal input.

The present study was undertaken to determine if the phrenic responses to vagally mediated inputs are also affected by focal cooling of the intermediate areas (IA) of the ventral medulla. Anesthetized, paralyzed cats whose vagi and carotid sinus nerves had been cut were studied. The IA were cooled focally with a thermostatically controlled thermode. When the IA were 40 degrees C, low intensity vagal stimulation caused inhibition of phrenic activity. The stimulus was also applied when IA were cooled to 30 and 20 degrees C. The magnitude of the inhibition was unaffected by the cooling. In another series of experiments, high intensity vagal stimulation was used. This led to an hyperpnea when IA were 40 degrees C. The magnitude of the response was much smaller when the test stimulus was given at lower IA temperatures. The effect of cooling IA on the phrenic response to mechanical stimulation of pulmonary stretch receptors and airway irritant receptors were also studied in cats with intact vagi. We found that the response to irritant receptor, but not to stretch receptor, stimulation was abolished by the cooling. We conclude that the intermediate areas are involved in the integration of afferent input from airway irritant receptors that reaches the respiratory controller via high threshold vagal afferents, but not involved in processing signals from pulmonary stretch receptors.

Excitatory and inhibitory effects of morphine on the intercostal-to-phrenic respiratory reflex

The purpose of this study was to determine the effect of intravenously administered morphine on the intercostal-to-phrenic reflex in spinal (C1) cats. The carotid and vertebral arteries were ligated. In addition a metal clamp was placed around the neck and tightened in order to occlude all blood flow to the brain. The animals were vagotomized, paralyzed and ventilated artificially. End-tidal PCO2 and body temperature were kept constant by means of servocontrollers. The intercostal-to-phrenic reflex was activated by rhythmic tapping of the lower thorax anteriorly with a metal bar. Three different doses of morphine were used. The smallest dose (1 mg/kg) caused a marked stimulation of phrenic activity that lasted for more than 5 min. A larger dose (10 mg/kg) had only a mild excitatory effect. A much larger dose (50 mg/kg), on the other hand, caused inhibition of evoked phrenic activity. Possible mechanisms involved in mediating the dose dependent effects of morphine on the intercostal-to-phrenic reflex are discussed.