Motor control of the costal and crural diaphragm--insights from transcranial magnetic stimulation in man

The costal and crural parts of the diaphragm differ in their embryological development and physiological function. It is not known if this is reflected in differences in their motor cortical representation. We compared the response of the costal and crural diaphragms using varying intensities of transcranial magnetic stimulation of the motor cortex at rest and during submaximal and maximal inspiratory efforts. The costal and crural motor evoked potential recruitment curves during submaximal inspiratory efforts were similar. The response to stimulation before, during and at 10 and 30 min after 44 consecutive maximal inspiratory efforts was also the same. Using paired stimulations to investigate intra-cortical facilitatory and inhibitory circuits we found no difference between the costal and crural response with varying interstimulus intervals, or when conditioning and test stimulus intensity were varied. We conclude that supraspinal control of the costal and crural diaphragm is identical during inspiratory tasks.     

Comparison of the motor discharge to the crural and costal diaphragm in the rat

We compared the efferent innervation of the crural and costal regions of the mammalian diaphragm with regard to axonal motor discharge patterns and conduction speeds. Recordings were obtained from single crural (233) and costal (133) phrenic motoneurones. Median conduction speeds, calculated by spike triggered averaging (13.7 ms(-1) crural and 11.8 ms(-1) costal), and frequency histograms of conduction speed were not statistically significantly different between the two populations (p=0.27: Mann-Whitney test and p=0.9: Kolmogorov-Smirnov test, respectively). There was no difference in the proportions of inspiratory, post-inspiratory or non-respiratory units encountered in the crural and costal phrenic branches. Units that lacked respiratory rhythm did not express cardiac rhythm and were insensitive to ganglion blockade. In conclusion, there were few differences noted between the two motor pools and this may be related to the fact that the rat does not differentially regulate its diaphragm during swallowing and is not an emetic species.     

急性高碳酸血症对膈肌功能的影响

本文用电刺激犬膈神经（ＰＮＳ）、记录经膈压（Ｐｄｉ）－刺激频率（Ｆ）曲线、分析自主呼吸膈肌肌电频谱（ＥＭＧｄｉ）的方法，研究急性ＣＯ２潴留对膈肌功能的影响，并探讨ｐＨ值在其中的作用。发现：１．急性ＣＯ２潴留使Ｐｄｉ进行性下降、Ｐｄｉ－Ｆ曲线右移，ＥＭＧｄｉ频谱改变；２．维持正常的ｐＨ值可减轻Ｐｄｉ、ＥＭＧｄｉ的改变。提示急性ＣＯ２潴留严重损害膈肌功能，ｐＨ的变化于其中起重要作用。     

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.     

Motor control of the diaphragm in anesthetized rabbits

Diaphragmatic regions are recruited in a specialized manner either as part of a central motor program during non-respiratory maneuvers, e.g. vomiting, or because of reflex responses, e.g. esophageal distension. Some studies in cats and dogs suggest that crural and costal diaphragm may be differentially activated also in response to respiratory stimuli from chemoreceptors or lung and chest wall mechanoreceptors. To verify whether this could occur also in other species, the EMG activity from the sternal, costoventral, costodorsal, and crural diaphragm was recorded in 42 anesthetized rabbits in response to various respiratory maneuvers, such as chemical stimulation, mechanical loading, lung volume and postural changes before and after vagotomy, or a non-respiratory maneuver such as esophageal distension. Regional activity was evaluated from timing of the raw EMG signal, and amplitude and shape of the moving average EMG. In all animals esophageal distension caused greater inhibition of the crural than sternal and costal diaphragm, whereas under all the other conditions differential diaphragmatic activation never occurred. These results indicate that in response to respiratory stimuli the rabbit diaphragm behaves as a single unit under the command of the central respiratory control system.     

Role of the diaphragm in trunk rotation in humans

The objectives of the present study were to test the hypothesis that the costal diaphragm contracts during ipsilateral rotation of the trunk and that such trunk rotation increases the motor output of the muscle during inspiration. Monopolar electrodes were inserted in the right costal hemidiaphragm in six subjects, and electromyographic (EMG) recordings were made during isometric rotation efforts of the trunk to the right ("ipsilateral rotation") and to the left ("contralateral rotation"). EMG activity was simultaneously recorded from the parasternal intercostal muscles on the right side. The parasternal intercostals were consistently active during ipsilateral rotation but silent during contralateral rotation. In contrast, the diaphragm was silent in the majority of rotations in either direction, and whenever diaphragm activity was recorded, it involved very few motor units. In addition, whereas parasternal inspiratory activity substantially increased during ipsilateral rotation and decreased during contralateral rotation, inspiratory activity in the diaphragm was essentially unaltered and the discharge frequency of single motor units in the muscle remained at 13-14 Hz in the different postures. It is concluded that 1) the diaphragm makes no significant contribution to trunk rotation and 2) even though the diaphragm and parasternal intercostals contract in a coordinated manner during resting breathing, the inspiratory output of the two muscles is affected differently by voluntary drive during trunk rotation.     

Effect of carbon dioxide on diaphragmatic function in human beings

We studied the effects of acute changes in the partial pressure of arterial carbon dioxide on diaphragmatic contractility and performance in four normal men. To study contractility we measured the ability of the diaphragm to generate pressure at a given level of excitation by determining the relation between the electrical activity of the diaphragm and transdiaphragmatic pressure during a voluntary quasi-isometric inspiratory effort carried out at different levels of end-tidal carbon dioxide. Our results show that contractility was reduced with hypercapnia (when end-tidal carbon dioxide was 7.5 per cent or higher), although hypocapnia (end-tidal carbon dioxide, 3 per cent) had no effect on diaphragmatic contractility. We also studied the development of diaphragmatic fatigue before and during carbon dioxide breathing. Subjects were studied at the same diaphragmatic tension-time index, a value analogous to the more familiar myocardial tension-time index, while the same inspiratory flow was maintained. Electromyographic signs of fatigue appeared at a lower tension-time index during hypercapnia than during normocapnia, indicating that endurance is diminished during hypercapnia. These findings show that acute respiratory acidosis equivalent to an arterial carbon dioxide tension of about 54 mm Hg decreases the contractility and endurance time of the diaphragm in human beings.     

Effect of salbutamol on respiratory muscle function and ventilation in awake canines

The effect of the beta-agonist bronchodilator salbutamol on respiratory muscles and ventilation is uncertain. The presence of beta2 receptors on skeletal muscles and increased diaphragm contractility in vitro with salbutamol predict a significant effect that has not been confirmed, in vivo in non-fatigued diaphragm or in clinical studies using standard bronchodilator dosages. Therefore, we infused salbutamol at a higher dosage (23.3 microg/min) used clinically for treatment of respiratory emergencies, while measuring directly the length, shortening and EMG activation of costal and crural diaphragm, parasternal intercostal and transversus abdominis muscles, in 10 awake canines. At this salbutamol dosage, ventilation and tidal volume increased significantly during both resting and CO2-stimulated breathing. Salbutamol elicited significant increases in respiratory muscle shortening with much smaller increases in EMG activity, so the proportionally greater muscle shortening per unit EMG showed increased muscle contractility. The effects of salbutamol were not extinguished by inspiratory flow resistance or fluid challenge but were reversed specifically by the beta-blocker, propranolol. This study demonstrates that, in sufficient intravenous dosage, the beta-agonist salbutamol elicits increased ventilation and a beta2 receptor-mediated increase in contractility of respiratory muscles.     

Differences in mouth occlusion pressure and breathing pattern between arm and leg incremental exercise

The aim of the study was to compare breathing pattern, mouth occlusion pressure, mean inspiratory flow and the ratio of mouth occlusion pressure to mean inspiratory flow at the same power output and carbon dioxide output during arm and leg incremental exercise. Mouth occlusion pressure was used as an index of inspiratory neuromuscular activity and its ratio to mean inspiratory flow as an index of the ‘effective’ impedance of the respiratory system. Eight normal subjects performed two incremental exercise tests, one with arms, the other with legs, on different weeks and in randomized order, and on two identical cycle ergometers. The power output was increased by steps of 25 W for arms and 50 W for legs every 4 min until exhaustion. At the same power output, oxygen consumption, carbon dioxide output, ventilation, mean inspiratory flow, mouth occlusion pressure, ‘effective’ impedance (P<0.001) and respiratory frequency (P<0.01) were higher during arm exercise than during leg exercise, whereas inspiratory time (P<0.05) and expiratory time (P<0.01) were lower. At the same carbon dioxide output, mouth occlusion pressure, ventilation, ‘effective’ impedance (P<0.001) and respiratory frequency (P<0.01) were higher and expiratory time (P<0.05) was lower during arm exercise. In conclusion, the higher inspiratory neuromuscular activity and impedance of the respiratory system during arm exercise and the differences observed in ventilation and breathing pattern at equal carbon dioxide output seem related to the differences in exercising muscle afferents and the presence of an increased load due to contraction of rib cage muscles to stabilize posture.     

急性缺氧对膈肌疲劳的影响

本文观察急性缺氧对犬膈肌疲劳的影响，发现急性缺氧条件下，膈肌疲劳的耐受时间为１８．００±３．７０ｍｉｎ，较对照组４１．３３±５．３０ｍｉｎ明显缩短（Ｐ＜０．０５），而且疲劳膈肌恢复过程中产生跨膈压（ｔｒａｎｓｄｉａｐｈｒａｇｍａｔｉｃｐｒｅｓｓｕｒｅ，Ｐｄｉ）明显低于对照组（Ｐ＜０．０５），证实急性缺氧使膈肌疲劳的耐受性明显下降，且不利于疲劳膈肌肌力的恢复。     

Activity patterns of the diaphragm during voluntary movements in awake cats

The diaphragm is an important inspiratory muscle, and is also known to participate in the postural function. However, the activity of the diaphragm during voluntary movements has not been fully investigated in awake animals. In order to investigate the diaphragmatic activity during voluntary movements such as extending or rotating their body, we analyzed the electromyogram (EMG) of the diaphragm and trunk muscles in the cat using a technique for simultaneous recordings of EMG signals and video images. Periodic respiratory discharges occurred in the left and right costal diaphragm when the cat kept still. However, once the cat moved, their periodicity and/or synchrony were sometimes buried by non-respiratory activity. Such non-periodic diaphragmatic activities during voluntary movements are considered as the combination of respiratory activity and non-respiratory activity. Most of the diaphragmatic activities started shortly after the initiation of standing-up movements and occurred after the onset of trunk muscle activities. Those activities were more active compared to the normal respiratory activity. During rotation movements, left and right diaphragmatic activities showed asymmetrical discharge patterns and higher discharges than those during the resting situation. This asymmetrical activity may be caused by taking different lengths of each side of the diaphragm and trunk muscles. During reaching movements, the diaphragmatic activity occurred prior to or with the onset of trunk muscle activities. It is likely that diaphragmatic activities during reaching movements and standing-up movements may have been controlled by some different control mechanisms of the central nervous system. This study will suggest that the diaphragmatic activity is regulated not only by the respiratory center but also by inputs from the center for voluntary movements and/or sensory reflex pathways under the awake condition.     

Diaphragmatic silent period to transcranial magnetic cortical stimulation for assessing cortical motor control of the diaphragm

This study was designed to determine whether a silent period could be elicited in the diaphragm electromyographic (EMG) activity by transcranial magnetic stimulation (TMS) of the motor cortex and, if so, to assess the influence of reflex or voluntary control of breathing on diaphragmatic cortical silent period (cSP). Diaphragmatic EMG activity was recorded in six healthy volunteers after motor cortex TMS triggered by the inspiratory flow peak and applied during forced inspiration (FI), voluntary hyperventilation (vHV) and reflex hyperventilation (rHV) to a CO(2) stimulus. Electrophysiological and respiratory parameters were studied, including diaphragmatic cSP duration and transdiaphragmatic pressure swing (DeltaPdi). A diaphragmatic cSP was found and correlated with DeltaPdi values. DeltaPdi and cSP duration were similar in the vHV and rHV conditions but were significantly increased during FI. This study established for the first time the existence of a diaphragmatic cSP to motor cortex TMS. The diaphragmatic cSP duration depended on the magnitude of the respiratory effort, as assessed by DeltaPdi, but not on the mechanism (volitional or reflex) of diaphragm activation.     

Myoelectric power spectrum dependence on muscular contraction level of elbow flexors

Summary The influence of the strength of contraction on surface recorded myoelectric power spectra was studied for three elbow flexors. Four subjects performed brief (3–5 s) isometric contraction levels (5–80% MVC). The experiment was repeated 23–26 times on different days. The surface myoelectric signal was recorded from the biceps brachii, the brachialis and the brachioradialis. By fast Fourier transform the myoelectric power spectrum was computed. The mean power frequency (MPF) was calculated and used as a single estimate of the myoelectric power spectrum. The MPF was found to increase with contraction strength with low level contractions. At levels in excess of 25–30% of MVC, the MPF became independent of contraction level. This dependence of the MPF on low level contractions is explained by tissue filtering effects and the recruitment order and distribution of motor units.This investigation was supported by grants from the Swedish Work Environment Fund and the Medical Faculty of Umeå University     

Effects of endurance training on myosin heavy-chain isoforms and enzyme activity in the rat diaphragm

We investigated the effects of endurance training (20 m/min, 60 min/day, 5 days/week) on myosin heavy-chain (MHC) isoforms and succinic dehydrogenase (SDH) activity in rat crural and costal diaphragms, and plantaris muscles. Although the 4-week endurance training produced significant (P<0.05) increases, both in SDH activity and the percentage of isoform HCIIa in the plantaris of the trained rat compared with the sedentary control rat, these alterations did not occur in either the crural or costal diaphragms. After 10 weeks of endurance training, trained animals had significantly (P<0.05) higher SDH activity in the costal diaphragm and the plantaris. Moreover, a significant (P<0.05) decrease occurred in the percentage of HCIIb in the costal diaphragm, and a significant (P<0.01) decrease in the percentage of HCIIb concomitant with a significant (P<0.05) increase of HCIIa resulted in the plantaris. However, the crural diaphragm did not show any significant changes after 10 weeks of endurance training. These results indicate that endurance training induces an alteration in the expression of an MHC phenotype, in addition to causing an increase in oxidative enzyme activity. However, the alterations in response to endurance training are apparently not uniform, varying between regions and/or kinds of muscles.     

Aminophylline increases ventilation and diaphragm contractility in awake canines

Traditional theophylline bronchodilators are still used clinically, especially in COPD. However, the effect of theophyllines on ventilation and respiratory muscles remains uncertain and these effects have not been measured directly in any awake, intact mammal. We hypothesized that aminophylline in the usual therapeutic dosage range, would elicit in the awake mammal, a significant increase in ventilation, and a significant increase in costal diaphragm shortening and contractility as recorded directly from the muscle. Therefore, we studied 13 awake canines, which had been chronically implanted with fine-wire EMG electrodes and sonomicrometer crystals in the costal segment of the diaphragm. Ventilatory parameters, moving average muscle EMG activity and muscle length and shortening, were measured at baseline and with aminophylline, during resting and hypercapnic stimulated breathing. Experiments were carried out prior to administration of aminophylline (baseline), and 1.5 h after loading and ongoing infusion with aminophylline. Minute ventilation, tidal volume and respiratory frequency all increased significantly with aminophylline, both during resting breathing and at equivalent levels of hypercapnic stimulated breathing. Costal diaphragm baseline muscle length was entirely unchanged with aminophylline. Costal diaphragm shortening increased significantly with aminophylline while corresponding costal diaphragm EMG activity remained constant, consistent with increased diaphragm contractility. Thus, in awake, intact mammals, aminophylline in usual therapeutic dosage elicits increased ventilation and increased contractility of respiratory muscles.     

Expiratory activity of the inspiratory muscles during cough.

To investigate the neural mechanism of the expiratory activity of the inspiratory muscles during a cough, EMG of the respiratory muscles were recorded in anesthetized and tracheostomized dogs. A laparoscope was used to minimize injury to the abdominal muscles for implantation of the electrodes into the costal diaphragm. During the expulsive phase of a cough, the diaphragm was active in 7 of 12 dogs and the external intercostal muscle was active in 3 of 6 dogs. During a cough, the expiratory activity of the diaphragm, after the termination of its inspiratory activity, started at 52.9 +/- 24.6 ms, and that of external intercostal muscle started at 51.1 +/- 20.5 ms. The expiratory activity of the internal intercostal muscle and of the transversus abdominis started at 34.3 +/- 13.0 and 27.8 +/- 15.2 ms, respectively. The onset of expiratory activity of the inspiratory muscles is significantly later than that of expiratory muscles. Continuous activity in the expiratory muscles evoked by airway occlusion, i.e., Hering-Breuer reflex, was suppressed during the inspiratory phase of a cough, but not suppressed during the expulsive phase even when the expiratory activity of the diaphragm was observed. We concluded that the expiratory activity of inspiratory muscles is controlled independently of both expiratory activity of the expiratory muscles and inspiratory activity of the inspiratory muscles.     

Compartmental chest wall volume changes during volitional normocapnic hyperpnoea

During increased ventilation, inspiratory rib cage muscles have been suggested to take over part of diaphragmatic work after the diaphragm fatigues. We investigated the extent to which this proposed change in muscle recruitment is associated with changes in the relative contribution of chest wall compartments to tidal volume (V(T)). Thirteen healthy subjects performed 1 h of fatiguing normocapnic hyperpnoea. Chest wall volumes were assessed by optoelectronic plethysmography. While breathing frequency increased (43±3 to 56±5 breaths min(-1), p=0.006) and V(T) decreased during normocapnic hyperpnoea (2.6±0.2 to 1.9±0.1l, p<0.001), the relative contribution of chest wall compartments to V(T) remained unchanged (pulmonary rib cage: 48±9 versus 51±14%; abdominal rib cage: 24±4 versus 23±9%; abdomen: 28±8 versus 26±9%; all p>0.05). In conclusion, fatiguing respiratory work is not associated with a change in compartmental contribution to V(T), even in the presence of a change in breathing pattern.     

Effects of theophylline on diaphragmatic strength and fatigue in patients with chronic obstructive pulmonary disease

We studied the effects of theophylline on diaphragmatic strength and fatigue in 15 patients with severe chronic obstructive pulmonary disease. Diaphragmatic strength was assessed by measurement of the transdiaphragmatic pressure generated at functional residual capacity during a maximal inspiratory effort against closed airways. Diaphragmatic fatigue was induced by resistive loaded breathing. The electrical activity of the diaphragm was recorded with an esophageal electrode during the fatigue runs, and the high-low ratio of the electrical signal was analyzed to assess diaphragmatic fatigue. Studies were performed before and after 7 and 30 days of theophylline administration (mean plasma level, 13 +/- 2 mg per liter). A control group received a placebo instead of theophylline. Theophylline increased maximal transdiaphragmatic pressure by 16 per cent after 7 days of administration (P less than 0.01), and this increase persisted after 30 days. No significant change in maximal transdiaphragmatic pressure was observed in the group given the placebo. Theophylline also suppressed diaphragmatic fatigue in all patients who received it. We conclude that theophylline has a potent and long-lasting effect on diaphragmatic strength and fatigue in patients with fixed airway obstruction.     

Central and peripheral components of diaphragmatic fatigue during inspiratory resistive load in cats

The development of fatigue was investigated in the diaphragm of anaesthetized, tracheostomized, spontaneously breathing cats during restricted air flow. Ventilation, transdiaphragmatic pressure (P_di), integrated electrical activity of diaphragm (E_di) and phrenic nerve (E_ph) were measured simultaneously and expressed as a percentage of values at unloaded breathing. Inspiratory loads were 60, 70 and 80% of P_{di max}. The P_{di max} was measured by airway occlusion at functional residual capacity. The duration of loads was 40–60 min. The diaphragmatic fatigue developed only during heavy inspiratory loading (80% P_{di max}). During the first 10 min of heavy load P_di, E_di and E_ph increased to 905 ± 60%, 248 ± 20% and 229 ± 24%, respectively (P < 0.01), and then began to fall gradually. Ventilation declined to 39 ± 3% after 60 min of heavy load (P < 0.01), resulting in acute hypercapnia and hypoxia. Initial fatigue appeared as a decrease in P_di (to 781 ± 63%) and parallel decline in E_di (to 233 ± 21%) after 30 min of load (P < 0.05). Phrenic nerve activity did not change during this stage. These data suggest a peripheral basis of diaphragmatic fatigue, related to disorders in neuromuscular transmission. After 60 min of heavy load, P_di fell to 675 ± 49%, E_di declined to 209 ± 28% and E_ph decreased to 189 ± 25%. We interpret the decrease in phrenic nerve activity as a weakening of central inspiratory drive and development of the central component of diaphragmatic fatigue in the last stage.     

Separate resistive loading of the respiratory phases during mild hypercapnia in man

Eleven human subjects were studied during steady state, controlled mild hypercapnia with resistive loading of either inspiration (RI) or expiration (RE). Minute ventilation and frequency were significantly reduced by RI (P = less than 0.01) and even more so by RE (P = less than 0.001). Tidal volume was unchanged. Both RI and RE reduced mean flow in the loaded phase - an effect relatively greater with RE. Neither RI nor RE altered mean flow in the unloaded phase. Although mean inspiratory flow was unchanged with RE, mouth occlusion pressure (P0.1) was increased (P = less than 0.01). Functional residual capacity (seven subjects) was increased with RE, but not with RI (P = less than 0.05). Five additional subjects were similarly studied with and without RE in whom transdiaphragmatic pressure (PDi) and peak diaphragmatic EMG (EMGDi) were examined. Changes in ventilation, breathing pattern and P0.1 were similar to those described above. Neither PDi nor EMGDi were significantly altered by RE, but with RE, diaphragmatic EMG activity began 50-190 ms before inspiratory flow. In conclusion, ventilation is reduced more by RE than by RI due to greater respiratory phase time. Moderately heavy RE does not augment inspiratory drive as reflected by mean flow, PDi or EMGDi. With RE and increased FRC, P0.1 does not accurately reflect inspiratory drive because of dissociation between EMG and flow.