Neuromuscular Coupling Research Articles

BAC transgenic mice express enhanced green fluorescent protein in central and peripheral cholinergic neurons

The peripheral nervous system has complex and intricate ramifications throughout many target organ systems. To date this system has not been effectively labeled by genetic markers, due largely to inadequate transcriptional specification by minimum promoter constructs. Here we describe transgenic mice in which enhanced green fluorescent protein (eGFP) is expressed under the control of endogenous choline acetyltransferase (ChAT) transcriptional regulatory elements, by knock-in of eGFP within a bacterial artificial chromosome (BAC) spanning the ChAT locus and expression of this construct as a transgene. eGFP is expressed in ChAT(BAC)-eGFP mice in central and peripheral cholinergic neurons, including cell bodies and processes of the somatic motor, somatic sensory, and parasympathetic nervous system in gastrointestinal, respiratory, urogenital, cardiovascular, and other peripheral organ systems. Individual epithelial cells and a subset of lymphocytes within the gastrointestinal and airway mucosa are also labeled, indicating genetic evidence of acetylcholine biosynthesis. Central and peripheral neurons were observed as early as 10.5 days postcoitus in the developing mouse embryo. ChAT(BAC)-eGFP mice allow excellent visualization of all cholinergic elements of the peripheral nervous system, including the submucosal enteric plexus, preganglionic autonomic nerves, and skeletal, cardiac, and smooth muscle neuromuscular junctions. These mice should be useful for in vivo studies of cholinergic neurotransmission and neuromuscular coupling. Moreover, this genetic strategy allows the selective expression and conditional inactivation of genes of interest in cholinergic nerves of the central nervous system and peripheral nervous system.

Therapeutic Cooling

Stretch reflexes contribute to joint stiffness, but the effects of therapeutic cooling on these reflexes are unknown. Therapeutic cooling is frequently used in knee rehabilitation, for instance, after rupture of the anterior cruciate ligament. Cooling a joint can affect nerve conduction velocity and the function of sensory organs. Such changes in neuromuscular coupling could reduce knee stability and increase the risk of knee injury. The aim of the present study was to evaluate whether there are negative effects of knee cooling on the hamstring short and medium latency reflex responses or on the anterior tibia trajectory after mechanically induced tibia translation. In 15 healthy volunteers, the latency and size of short latency and medium latency reflex responses of hamstring muscles were assessed before and after 20 min of cold therapy of the knee joint as applied in rehabilitation. Reflex responses were evoked by a mechanically induced posterior-anterior tibia translation during standing. Reflexes were recorded by surface electromyography. The distance of anterior tibia motion and its velocity were assessed by potentiometric position transducer. Local cold therapy of the knee did not alter the latency or the size of short or medium latency responses. Also, the extent and the velocity of tibia translation were unchanged after knee cooling. Cold therapy does not seem to adversely influence the spinal reflexes of the hamstrings induced by anterior tibia translation. As neuromuscular coupling was not significantly affected, the cold therapy, as performed in the present study, is not likely to increase the risk of knee injury.

Potential Advantages of Patient–Ventilator Synchrony

During conventional mechanical ventilation, fixed set pressure, flow, and tidal volume result in a mismatch between patient and ventilator inspiratory time and in a patient's inability to adapt to changing ventilatory demand. Synchrony between the patient and ventilator improves neuromuscular coupling and the ability to adapt to increased ventilatory demand or loading. The sensation of dyspnea prevents ineffective inspiratory efforts and attenuates periodic breathing during sleep.

Interaction of neuromuscular, spatial and visual constraints on hand–foot coordination dynamics

In the present study, we investigated the contributions of motor and perceptual processes to directional constraints as observed during hand–foot coordination. Participants performed cyclical flexion–extension movements of the right hand and foot under two coordination modes: in-phase (isodirectional) and antiphase (non-isodirectional). Those tasks were performed either with full vision or no vision of the limbs. Depending on the position of the forearm (prone or supine), the coordination patterns were performed with similar and dissimilar neuro-muscular coupling with respect to their phylogenetic origin as antigravity muscles. Results showed that the antiphase pattern was more difficult to maintain than the in-phase pattern and that neuro-muscular coupling significantly influenced the coordination dynamics. Moreover, the effect of vision differed as a function of both neuro-muscular coupling and coordination mode. Under dissimilar neuro-muscular coupling, the presence of visual feedback stabilized the in-phase pattern and destabilized the antiphase pattern. In contrast, visual feedback did not influence pattern stability during conditions of similar neuro-muscular coupling. These results shed light on the complex interactions between motor and perceptual (visual) constraints during the production of hand–foot coordination patterns.

Exercise Training Improves Exertional Dyspnea in Patients With COPD : Evidence of the Role of Mechanical Factors

To our knowledge, no data have been reported on the effects of exercise training (EXT) on central respiratory motor output or neuromuscular coupling (NMC) of the ventilatory pump, and their potential association with exertional dyspnea. Accurate assessment of these important clinical outcomes is integral to effective management of breathlessness of patients with COPD. Twenty consecutive patients with stable moderate-to-severe COPD were tested at 6-week intervals at baseline, after a nonintervention control period (pre-EXT), and after EXT. Patients entered an outpatient pulmonary rehabilitation program involving regular exercise on a bicycle. Incremental symptom-limited exercise testing (1-min increments of 10 W) was performed on an electronically braked cycle ergometer. Oxygen uptake (O(2)), carbon dioxide output (CO(2)), minute ventilation (E), time, and volume components of the respiratory cycle and, in six patients, esophageal pressure swings (Pessw), both as actual values and as percentage of maximal (most negative in sign) esophageal pressure during sniff maneuver (Pessn), were measured continuously over the runs. Exertional dyspnea and leg effort were evaluated by administering a Borg scale. Measurements at baseline and pre-EXT were similar. Significant increase in exercise capacity was found in response to EXT: (1) peak work rate (WR), O(2), CO(2), E, tidal volume (VT), and heart rate increased, while peak exertional dyspnea and leg effort did not significantly change; (2) exertional dyspnea/O(2) and exertional dyspnea/CO(2) decreased while E/O(2) and E/CO(2) remained unchanged. The slope of both exertional dyspnea and leg effort relative to E fell significantly after EXT; (3) at standardized WR, E, and CO(2), exertional dyspnea and leg effort decreased while inspiratory capacity (IC) increased. Decrease in E was accomplished primarily by decrease in respiratory rate (RR) and increase in both inspiratory time (TI) and expiratory time; VT slightly increased, while inspiratory drive (VT/TI) and duty cycle (TI/total time of the respiratory cycle) remained unchanged. The decrease in Pessw and the increase in VT were associated with lower exertional dyspnea after EXT; (4) at standardized E, VT, RR, and IC, Pessw and Pessw(%Pessn)/VT remained unchanged while exertional dyspnea and leg effort decreased with EXT. In conclusion, increases in NMC, aerobic capacity, and tolerance to dyspnogenic stimuli and possibly breathing retraining are likely to contribute to the relief of both exertional dyspnea and leg effort after EXT.

Control of Breathing in Patients with Chronic Pulmonary Obstructive Disease: Response to Bamiphylline

In two groups (A and B) of patients with severe chronic obstructive pulmonary disease (COPD), matched for age and routine pulmonary function testing, we evaluated inspiratory muscle strength (MIP), breathing pattern, mouth occlusion pressure (P0.1), inspiratory neural drive, assessed in terms of electromyographic activity of both diaphragm (EMGd) and intercostal (EMGint) muscles, and P0.1/EMGd ratio, an index of inspiratory neuromuscular coupling. Group A (8 patients) was studied before and after a 7-day period of a new oral xanthine derivative (bamiphylline, 1.2 g daily), and group B (7 patients) was given a placebo. Under control conditions, compared with an age-matched normal control group, groups A and B both exhibited a decrease in MIP, more rapid (increase in respiratory frequency RF) and shallower (decrease in tidal volume; VT) breathing (RSB), a marked increase in both EMGd and EMGint and a lower P0.1/EMGd ratio. With bamiphylline FEV1 and PaO2 significantly increased, while a substantial increase in MIP was found in 5 out of the 8 patients. VT and inspiratory time (Ti) also significantly increased, and RF decreased. Both EMGd and EMGint significantly decreased, whereas P0.1/EMGd exhibited a substantial increase in 5 out of the 8 patients. Conversely, no significant changes were observed in group B during the study period. From these data we conclude that in patients with COPD, clinical manifestations, probably associated with inspiratory muscle overloading (decrease in muscle strength, RSB, increase in respiratory neural drive, and derangement in neuromuscular coupling) may benefit from a short-term treatment with bamiphylline.

Neural Respiratory Drive and Neuromuscular Coupling in Patients with Chronic Obstructive Pulmonary Disease (COPD)

In 15 spontaneously breathing patients with chronic obstructive pulmonary disease (COPD) divided into two groups, one with normocapnia (A) and one with chronic hypercapnia (B), we evaluated the maximal voluntary inspiratory muscle strength (MIP), the pattern of breathing, the mouth occlusion pressure (Po.1), the neural respiratory drive (NRD), assessed by surface electromyographic (EMG) activity of the diaphragm (EMGd) and EMG activity of intercostal muscles (EMGint), and the chest wall neuromuscular coupling, assessed in terms of Po.1/EMGd ratio. Compared with an age-matched normal control group, both A and B groups exhibited lower MIP, significantly greater EMGd and EMGint, and lower Po.1/EMGd ratio. However, a similar pattern, along with a rapid and shallow breathing, differentiated group B from group A. In group B we found a significant direct relationship between Po.1/EMGd ratio and MIP, and an inverse relationship between PaCO2 and Po.1/EMGd ratio. These data seem to indicate the following: (1) EMG is a more precise method than Po.1 in assessing the magnitude of the NRD; (2) NRD is increased in these patients; and (3) clinical manifestations probably associated with inspiratory muscle fatigue (marked decrease in muscle strength, rapid and shallow breathing, and alveolar hypoventilation) may be accompanied by a greater NRD and a more marked derangement in chest wall neuromuscular coupling in COPD.

Neural Respiratory Drive and Neuromuscular Coupling During CO2 Rebreathing in Patients with Chronic Interstitial Lung Disease

In 12 patients with CILD and 18 age-matched normal subjects we assessed the ventilatory control system at three levels: (a) neural, as assessed by EMGd (XP/Ti) and EMGint muscles via surface electrodes; (b) muscular, as assessed by mouth occlusion pressure (P0.1); and (c) ventilatory, as assessed by both ventilation (VE) and the related parameters, tidal volume (VT) and respiratory frequency (f). Compared with a normal control group, patients exhibited a significant decrease in lung volumes and in MIP; VT and inspiratory time (Ti) were significantly lower, while VT/Ti, P0.1, and both EMGd and EMGint were significantly greater in patients. During a CO2 rebreathing test, patients exhibited significantly greater EMGd, EMGint, and P0.1 responses to increasing PETCO2 than the control group. VE response slopes were similar in the two groups. For a given EMGd response slope (delta XP/Ti/delta PETCO2), the average P0.1 response slope (delta P0.1/delta PETCO2) was found to be significantly lower in patients than in the normal control group. Compared with normal subjects, CILD patients have a normal or increased neural component of respiratory activity and relatively low neuromuscular coupling (delta P0.1/delta XP/Ti). The decreased neuromuscular coupling could be explained in these patients by a reduced inspiratory muscle strength.

Changes in neural drive (EMGd) and neuromuscular coupling during histamine-induced bronchoconstriction in patients with asthma

This study was undertaken in order to assess the neural drive to the respiratory muscles and the inspiratory neuromuscular coupling in patients with bronchial asthma during histamine-induced bronchoconstriction. Bronchoconstriction was produced in a graded fashion, with histamine phosphate aerosol of increasing dose, in twelve asymptomatic asthmatic patients and was measured by FEV1. Inspiratory drive was measured by electromyographic activity of the diaphragm (EMGd) and the coupling of the neural drive to the respiratory muscles was assessed by the relationship of mouth occlusion pressure (P0.1) to EMGd. During the test we also measured electromyographic activity of the inspiratory intercostal (EMGint), sternomastoid (EMGsm) and expiratory abdominal (EMGab) muscles. Histamine caused a significant decrease in FEV1, a significant increase in P0.1, EMGd, EMGint, and a relevant increase in EMGsm, with no substantial increase in EMGab. An inverse significant relationship between the change in FEV1 and changes in P0.1, EMGd and EMGint and a significant correlation between the change in FEV1 and in the P0.1/EMGd ratio were observed. We conclude that a progressive increase in bronchospasm is accompanied by a progressive increase in respiratory neural drive and decrease in neuromuscular coupling. This could be caused both by an increase in lung volume and a lack of abdominal expiratory muscle recruitment.

Effects of aminophylline on respiratory drive and neuromuscular coupling in normal man and in patients with chronic airflow obstruction.

In order to evaluate the separate effects of aminophylline on the neural and muscular components of the respiratory control system, assessed by electromyographic activity of the diaphragm (EMGd) and mouth occlusion pressure (P0.1), respectively, 6 normal subjects and 14 patients with mild or moderate chronic airflow obstruction (8 asthmatics and 6 COPD) were studied during CO2 rebreathing, before and after administration of a therapeutic dose of aminophylline 5.6 mg/kg. Compared to normal subjects, before aminophylline administration both asthmatic and COPD patients exhibited a significantly greater value in EMGd response slope to CO2. In no group did aminophylline modify P0.1 or EMGd activity response slope to CO2, nor did it significantly affect neuromuscular coupling, assessed by plotting change in P0.1 against change in EMGd activity with increasing CO2. The data appear to indicate that aminophylline in therapeutic concentrations does not modify respiratory drive or neuromuscular coupling in normal subjects, or in patients with mild or moderate chronic airflow obstruction.

Respiratory muscle function during CO2 rebreathing with inspiratory flow-resistive loading.

We investigated the respiratory muscle contribution to inspiratory load compensation by measuring diaphragmatic and intercostal electromyograms (EMGdi and EMGic), transdiaphragmatic pressure (Pdi), and thoracoabdominal motion during CO2 rebreathing with and without 15 cmH2O X l-1 X s inspiratory flow resistance (IRL) in normal sitting volunteers. During IRL compared with control, Pdi measured during airflow and during airway occlusion increased for a given change in CO2 partial pressure and EMGdi, and there was a greater decrease in abdominal (AB) end expiratory anteroposterior dimensions with increased expiratory gastric pressure (Pga), this leading to an inspiratory decline in Pga with outward AB movement, indicating a passive component to the descent of the abdomen-diaphragm. The response of EMGic to IRL was similar to that of EMGdi, though rib cage (RC)-Pga plots did infer intercostal muscle contribution. We conclude that during CO2 rebreathing with IRL there is improved diaphragmatic neuromuscular coupling, the prolongation of inspiration promoting a force-velocity advantage, and increased AB action serving to optimize diaphragm length and configuration, as well as to provide its own passive inspiratory action. Intercostal action provides increased assistance also. Therefore, compensation for inspiratory resistive loads results from the combined and integrated effort of all respiratory muscle groups.