Animal models for genetic neuromuscular diseases

The neuromuscular disorders are a heterogeneous group of genetic diseases, caused by mutations in genes coding sarcolemmal, sarcomeric, and citosolic muscle proteins. Deficiencies or loss of function of these proteins leads to variable degree of progressive loss of motor ability. Several animal models, manifesting phenotypes observed in neuromuscular diseases, have been identified in nature or generated in laboratory. These models generally present physiological alterations observed in human patients and can be used as important tools for genetic, clinic, and histopathological studies. The mdx mouse is the most widely used animal model for Duchenne muscular dystrophy (DMD). Although it is a good genetic and biochemical model, presenting total deficiency of the protein dystrophin in the muscle, this mouse is not useful for clinical trials because of its very mild phenotype. The canine golden retriever MD model represents a more clinically similar model of DMD due to its larger size and significant muscle weakness. Autosomal recessive limb-girdle MD forms models include the SJL/J mice, which develop a spontaneous myopathy resulting from a mutation in the Dysferlin gene, being a model for LGMD2B. For the human sarcoglycanopahties (SG), the BIO14.6 hamster is the spontaneous animal model for delta-SG deficiency, whereas some canine models with deficiency of SG proteins have also been identified. More recently, using the homologous recombination technique in embryonic stem cell, several mouse models have been developed with null mutations in each one of the four SG genes. All sarcoglycan-null animals display a progressive muscular dystrophy of variable severity and share the property of a significant secondary reduction in the expression of the other members of the sarcoglycan subcomplex and other components of the Dystrophin-glycoprotein complex. Mouse models for congenital MD include the dy/dy (dystrophia-muscularis) mouse and the allelic mutant dy(2J)/dy(2J) mouse, both presenting significant reduction of alpha 2-laminin in the muscle and a severe phenotype. The myodystrophy mouse (Large(myd)) harbors a mutation in the glycosyltransferase Large, which leads to altered glycosylation of alpha-DG, and also a severe phenotype. Other informative models for muscle proteins include the knockout mouse for myostatin, which demonstrated that this protein is a negative regulator of muscle growth. Additionally, the stress syndrome in pigs, caused by mutations in the porcine RYR1 gene, helped to localize the gene causing malignant hypertermia and Central Core myopathy in humans. The study of animal models for genetic diseases, in spite of the existence of differences in some phenotypes, can provide important clues to the understanding of the pathogenesis of these disorders and are also very valuable for testing strategies for therapeutic approaches.

Neuromuscular electrical stimulation improves GLUT-4 and morphological characteristics of skeletal muscle in rats with heart failure

Aim: Changes in skeletal muscle morphology and metabolism are associated with limited functional capacity in heart failure, which can be attenuated by neuromuscular electrical stimulation (ES). The purpose of the present study was to analyse the effects of ES upon GLUT-4 protein content, fibre structure and vessel density of the skeletal muscle in a rat model of HF subsequent to myocardial infarction. Methods: Forty-four male Wistar rats were assigned to one of four groups: sham (S), sham submitted to ES (S+ES), heart failure (HF) and heart failure submitted to ES (HF+ES). The rats in the ES groups were submitted to ES of the left leg during 20 days (2.5 kHz, once a day, 30 min, duty cycle 50%- 15 s contraction/15 s rest). After this period, the left tibialis anterior muscle was collected from all the rats for analysis. Results: HF+ES rats showed lower values of lung congestion when compared with HF rats (P = 0.0001). Although muscle weight was lower in HF rats than in the S group, thus indicating hypotrophy, 20 days of ES led to their recovery (P < 0.0001). In both groups submitted to ES, there was an increase in muscle vessel density (P < 0.04). Additionally, heart failure determined a 49% reduction in GLUT-4 protein content (P < 0.03), which was recovered by ES (P < 0.01). Conclusion: In heart failure, ES improves morphological changes and raises GLUT-4 content in skeletal muscle.

Relative contribution of pre- and post-synaptic effects to the neostigmine-induced recovery of neuromuscular transmission blocked by vecuronium

The relative contribution of the pre- and post-synaptic effects to the neostigmine-induced recovery of neuromuscular transmission blocked by vecuronium was studied. A conjunction of myographical and electrophysiological techniques was employed. The preparation was the sciatic nerve-extensor digitorum longus muscle of the rat, in vitro. The physiological variables recorded were nerve-evoked twitches (generated at 0.1 Hz), tetanic contractions (generated at 50 Hz) and end-plate potentials (epps), generated in trains of 50 Hz. The epps were analyzed in: amplitude of first epp in the train; mean amplitude of the 30th to the 59th epp in the train (epps-plateau); half-decay time of the epp; early tetanic rundown of epps in the train; plateau tetanic rundown of epps in the train; quantal content of the epps and quantal size. In myographical experiments, a concentration of vecuronium was found (0.8 mu m) that affected both twitches and tetanic contractions and a concentration of neostigmine was found (0.048 mu m) that completely restored the twitch affected by vecuronium. The cellular effects of vecuronium and neostigmine, studied alone or in association, in the above-mentioned concentrations, were scrutinized by means of electrophysiological techniques. These showed that vecuronium alone decreased the peak amplitude, the quantal content of epps and the quantal size and reinforced the tetanic rundown of epps. Neostigmine alone increased the peak amplitude, the quantal content and the half-decay time of the epps. When employed in the presence of vecuronium, neostigmine increased both the quantal content of the epps (via a presynaptic effect) and the half-decay time of the epps (via a postsynaptic effect). Seeing the pre- and the post-synaptic effects of neostigmine were of similar magnitude, they permit to conclude that both these effects contributed significantly to the restoration by neostigmine of the neuromuscular transmission blocked by vecuronium.

Differential effects of neuromuscular blockers on twitches and tetani in the isolated rat muscle: a multiple comparison study using simultaneous confidence intervals

In the present work a comparative quantitative evaluation of the differential effects of neuromuscular blockers on twitches and tetani was performed, encompassing: atracurium, cisatracurium, mivacurium, pancuronium, rocuronium and vecuronium. The sciatic nerve-extensor digitorum longus muscle of the rat was used, in vitro. Twitches were evoked at 0.1 Hz and tetani at 50 Hz. The differential effects of the studied compounds on twitches and tetani were statistically compared using simultaneous confidence intervals for the ratios between mean IC(50) for the block of twitches and mean IC(50) for the block of tetani. The results of ratios of mean IC(50) together with their corresponding 95% simultaneous confidence intervals were: vecuronium: 2.5 (1.8-3.5); mivacurium: 3.8 (3.0-4.9); pancuronium: 3.9 (2.0-7.6); rocuronium: 6.1 (3.8-9.9); atracurium: 9.0 (6.4-12.6); cisatracurium: 13.1 (6.0-28.4). Using the criteria that neuromuscular blockers displaying disjunct confidence intervals for the ratios of mean IC(50) differ statistically with regard to differential effects on twitches and tetani, significant differences in ratios of IC(50) were detected in the following cases: vecuronium vs. rocuronium, vs. atracurium and vs. cisatracurium and mivacurium vs: cisatracurium and vs. atracurium. The results show that the magnitude of the differential effects of neuromuscular blockers on twitches and tetani, as evaluated in the present work in the form of ratios of mean IC(50), does not depend on the chemical structure (comparing steroidal and isoquinolinic compounds), but seems to depend on differential pre- and post-synaptic effects of the compounds. It is also suggested that the greater the ability of a compound to block twitches and tetani in a differential manner, the safer is the compound from the clinical anesthesiology viewpoint.

Effect of neuromuscular electrical stimulation and isotonic exercises in flexor and extensor muscles of knee of hemiplegic patients

Aim. To verify the muscular force and resistance to the movement of the flexor and extensor muscles of the knee of patients with spasticity after treatment with neuromuscular electrical stimulation (NMES) and isotonic exercises. Patients and methods. The patients this study were divided into group I (NMES) and group 2 (isotonic exercises). Their muscular torque and resistance to the movement of the flexor and extensor knee muscles were measured by the isokinetic dynamometer and the degree of spasticity by the modified Ashworth scale before and after ten sessions. Results. Alterations in the scores of the modified Ashworth scale were not observed. An increase in the flexor torque in group 1 (p = 0.041) and in group 2 (p = 0.001) was verified. In the passive mode, group 1 presented a reduction of resistance to the flexion movement (p = 0.026), while in group 2, a reduction of resistance to both the flexion (p = 0,029) and extension movements (p = 0.019) was verified. Conclusions. The two therapeutical resources had their efficiency proven only for the increase of the force of the flexor muscles. The resistance to movement, the isotonic exercises were more effective because they promoted a reduction in the resistance of the flexor and extensor knee muscles.

Differences in the neuromuscular capacity and lean muscle tissue in old and older community-dwelling adults

This study investigated whether there was a worsening of the neuromuscular capacity of older adults after the seventh decade of life. The results suggest that the age-related deterioration in maximal strength measures and rapid force production characteristics in older adults could be related to a reduction in the mass and neural activation of the thigh muscles.

Efficacy of neuromuscular electrical stimulation in improving ankle kinetics during walking in children with cerebral palsy

Neuromuscular electrical stimulation (NMES) applied to the triceps surae muscle is claimed to be effective in improving gait in children with cerebral palsy. The main aim of this study was to determine the effect of NMES on the triceps surae muscle in improving the gait and function of children with cerebral palsy. Twelve children with spastic diplegia or hemiplegia were recruited and randomly assigned to the two experimental groups. The period of the study was 8 weeks (2-4-2 week design). The initial 2 weeks was the control period, in which usual treatment was given to both groups of patients with a pre- and post-treatment assessment. The middle 4 weeks was the experimental period, in which the Treadmill+NMES group received NMES plus treadmill walking training and the Treadmill group underwent treadmill walking training only. Assessment was performed at 2-week intervals. The final 2 weeks was the carryover period, in which treatment to be tested was stopped and reassessment performed again at the end of week 8. An additional treatment and post-treatment assessment were given at weeks 2, 4 and 6 to test for the immediate effect of treatment. Altogether, eight repeated measures with three-dimensional gait analysis and five clinical measurements using the gross motor function measure (GMFM) were performed. Kinetic changes in ankle moment quotient (AMQ) and ankle power quotient (APQ) were not significant either immediately or cumulatively in both groups. Improvement in trend was observed in both groups immediately but not cumulatively. Using the GMFM, functional changes were detected in standing (GMST, p < 0.001) and in walking (GMWK, p = 0.003) using a 'time' comparison. Significant interaction was also detected in GMWK using 'treatment by time' (p = 0.035). The difference between the two groups was not significant on 'treatment' comparison of both GMST and GMWK. Both groups showed improvement in GMST and GMWK cumulatively but there was no difference between the two groups. The effects in both groups could be carried over to 2 weeks after interventions stopped. Both the Treadmill+NMES and Treadmill groups showed improvement in functional outcomes. The trend in the changes of the GMFM score suggested that improvements were greater in the Treadmill+NMES group. There was also a trend showing some immediate improvement in AMQ and APQ.

Older adults' neuromuscular adaptations to resistence training and effects on challenging gait tasks

Community locomotion is threatened when older individuals are required to negotiate obstacles, which place considerable stress on the musculoskeletal system. The vulnerability of older adults during challenging locomotor tasks is further compromised by age-related strength decline and muscle atrophy. The first study in this investigation determined the relationship between the major muscle groups of the lower body and challenging locomotor tasks commonly found in the community environment of older adults. Twenty-nine females and sixteen males aged between 62 and 88 years old (68.2 ±6.5) were tested for the maximal voluntary contraction (MVC) strength of the knee extensors and 1-RM for the hip extensors, flexors, adductors, abductors, knee extensors and flexors and ankle plantar flexors. Temporal measurements of an obstacle course comprising four gait tasks set at three challenging levels were taken. The relationship between strength and the obstacle course dependent measures was explored using linear regression models. Significant associations (p≤0.05) between all the strength measures and the gait performances were found. The correlation values between strength and obstructed gait (r = 0.356-0.554) and the percentage of the variance explained by strength (R2 = 13%-31%), increased as a function of the challenging levels, especially for the stepping over and on and off conditions. While the difficulty of community older adults to negotiate obstacles cannot be attributed to a single causal pathway, the findings of the first study showed that strength is a critical requirement. That the magnitude of the association increased as a function of the challenging levels, suggests that interventions aimed at improving strength would potentially be effective in helping community older adults to negotiate environmental gait challenges. In view of the findings of the first study, a second investigation determined the effectiveness of a progressive resistance-training program on obstructed gait tasks measured under specific laboratory conditions and on an obstacle course mimicking a number of environmental challenges. The time courses of strength gains and neuromuscular mechanisms underpinning the exercise-induced strength improvements in community-dwelling older adults were also investigated. The obstructed gait conditions included stepping over an obstacle, on and off a raised surface, across an obstacle and foot targeting. Forty-three community-living adults with a mean age of 68 years (control =14 and experimental=29) completed a 24-week progressive resistance training program designed to improve strength and induce hypertrophy in the major muscles of the lower body. Specific laboratory gait kinetics and kinematics and temporal measures taken on the obstacle course were measured. Lean tissue mass and muscle activation of the lower body muscle groups were assessed. The MVC strength of the knee extensors and 1-RM of the hip extension, hip flexion, knee extension, knee flexion and ankle plantar flexion were measured. A 25% increase on the MVC of the knee extensors (p≤0.05) was reported in the training group. Gains ranging between 197% and 285% were recorded for the 1-RM exercises in the trained subjects with significant improvements found throughout the study (p≤0.05). The exercise-induced strength gains were mediated by hypertrophic and neural factors as shown by 8.7% and 27.7% increases (p≤0.05) in lean tissue mass and integrated electromyographic activity, respectively. Strength gains were accompanied by increases in crossing velocity, stride length and reductions in stride duration, stance and swing time for all gait tasks except for the foot targeting condition. Specific kinematic variables associated with safe obstacle traverse such as vertical obstacle heel clearance, limb flexion, horizontal foot placements prior to and at post obstacle crossing and landing velocities resulted in an improved crossing strategy in the experimental subjects. Significant increases in the vertical and anterior-posterior ground reaction forces accompanied the changes in the gait variables. While further long-term prospective studies of falls rates would be needed to confirm the benefits of lower limb enhanced strength, the findings of the present study provide conclusive evidence of significant improvements to gait efficiency associated with a systematic resistance-training program. It appears, however, that enhanced lower body strength has limited effects on gait tasks involving a dynamic balance component. In addition, due to the larger strength-induced increases in voluntary activation of the leg muscle compared to relatively smaller gains in lean tissue mass, neural adaptations appear to play a greater contributing role in explaining strength gains during the current resistance training protocol.

Neuromuscular control and running economy is preserved in elite international triathletes after cycling

Running is the most important discipline for Olympic triathlon success. However, cycling impairs running muscle recruitment and performance in some highly trained triathletes; though it is not known if this occurs in elite international triathletes. The purpose of this study was to investigate the effect of cycling in two different protocols on running economy and neuromuscular control in elite international triathletes. Muscle recruitment and sagittal plane joint angles of the left lower extremity and running economy were compared between control (no preceding cycle) and transition (preceded by cycling) runs for two different cycle protocols (20-minute low-intensity and 50-minute high-intensity cycles) in seven elite international triathletes. Muscle recruitment and joint angles were not different between control and transition runs for either cycle protocols. Running economy was also not different between control and transition runs for the ow-intensity (62.4 ^ 4.5 vs. 62.1 ^ 4.0 ml/min/kg, p . 0.05) and high-intensity (63.4 ^ 3.5 vs. 63.3 ^ 4.3 ml/min/kg, p . 0.05) cycle protocols. The results of this study demonstrate that both low- and high-intensity cycles do not adversely influence neuromuscular control and running economy in elite international triathletes.

Neuromuscular adaptations to training, injury and passive interventions: implications for running economy

Performance in endurance sports such as running, cycling and triathlon has long been investigated from a physiological perspective. A strong relationship between running economy and distance running performance is well established in the literature. From this established base, improvements in running economy have traditionally been achieved through endurance training. More recently, research has demonstrated short-term resistance and plyometric training has resulted in enhanced running economy. This improvement in running economy has been hypothesized to be a result of enhanced neuromuscular characteristics such as improved muscle power development and more efficient use of stored elastic energy during running. Changes in indirect measures of neuromuscular control (i.e. stance phase contact times, maximal forward jumps) have been used to support this hypothesis. These results suggest that neuromuscular adaptations in response to training (i.e. neuromuscular learning effects) are an important contributor to enhancements in running economy. However, there is no direct evidence to suggest that these adaptations translate into more efficient muscle recruitment patterns during running. Optimization of training and run performance may be facilitated through direct investigation of muscle recruitment patterns before and after training interventions.

There is emerging evidence that demonstrates neuromuscular adaptations during running and cycling vary with training status. Highly trained runners and cyclists display more refined patterns of muscle recruitment than their novice counterparts. In contrast, interference with motor learning and neuromuscular adaptation may occur as a result of ongoing multidiscipline training (e.g. triathlon). In the sport of triathlon, impairments in running economy are frequently observed after cycling. This impairment is related mainly to physiological stress, but an alteration in lower limb muscle coordination during running after cycling has also been observed. Muscle activity during running after cycling has yet to be fully investigated, and to date, the effect of alterations in muscle coordination on running economy is largely unknown. Stretching, which is another mode of training, may induce acute neuromuscular effects but does not appear to alter running economy.

There are also factors other than training structure that may influence running economy and neuromuscular adaptations. For example, passive interventions such as shoes and in-shoe orthoses, as well as the presence of musculoskeletal injury, may be considered important modulators of neuromuscular control and run performance. Alterations in muscle activity and running economy have been reported with different shoes and in-shoe orthoses; however, these changes appear to be subject-specific and nonsystematic. Musculoskeletal injury has been associated with modifications in lower limb neuromuscular control, which may persist well after an athlete has returned to activity. The influence of changes in neuromuscular control as a result of injury on running economy has yet to be examined thoroughly, and should be considered in future experimental design and training analysis.

Neuromuscular performance and bone structural characteristics in young healthy men and women

Muscle mass and strength have been shown to be important factors in bone strength. Low muscular force predisposes to falling especially among elderly. Regular exercise helps to prevent falls and resulting bone fractures. Better understanding of muscle function and its importance on bone properties may thus add information to fracture prevention. Therefore the purpose of this study was to examine the relationship between bone strength and muscular force production. Twenty-young men [24 (2) years] and 20 [24 (3) years] women served as subjects. Bone compressive (BSId) and bending strength indices (50 Imax) were measured with peripheral quantitative computed tomography (pQCT) at tibial mid-shaft and at distal tibia. Ankle plantarflexor muscle volume (MV) was estimated from muscle thickness measured with ultrasonography. Neuromuscular performance was evaluated from the measurements of maximal ground reaction force (GRF) in bilateral jumping and of eccentric maximal voluntary ankle plantarflexor torque (MVC). Specific tension (ST) of the plantarflexors was calculated by dividing the MVC with the muscle volume. Activation level (AL) was measured with superimposed twitch method. Distal tibia BSId and tibial mid-shaft 50 Imax correlated positively with GRF, MVC and MV in men (r = 0.45–0.67, P\0.05). Tibial mid-shaft 50 Imax and neuromuscular performance variables were correlated in women (r = 0.46–0.59, P\0.05), whereas no correlation was seen in distal tibia. In the regression analysis, MV and ST could explain 64% of the variance in tibial mid-shaft bone strength and 41% of the variation in distal tibia bone strength. The study emphasizes that tibial strength is related to maximal neuromuscular performance. In addition, tibial mid-shaft seems to be more dependent on the neuromuscular performance, than distal tibia. In young adults, the association between bone adaptation and neuromuscular performance seems to be moderate and also site and loading specific.