Neuromuscular Junction Damage Research Articles

BI1 Activates Autophagy and Mediates TDP43 to Regulate ALS Pathogenesis.

Amyotrophic lateral sclerosis (ALS) is the most prevalent motor neuron disease in adults. Currently, there are no known drugs or clinical approaches that have demonstrated efficacy in treating ALS. Mitochondrial function and autophagy have been identified as crucial mechanisms in the development of ALS. While Bax inhibitor 1 (BI1) has been implicated in neurodegenerative diseases, its exact mechanism remains unknown. This study investigates the therapeutic impact of BI1 overexpression on ALS both in vivo and in vitro, revealing its ability to mitigate SOD1G93A-induced apoptosis, nuclear damage, mitochondrial dysfunction, and axonal degeneration of motor neurons. At the same time, BI1 prolongs onset time and lifespan of ALS mice, improves motor function, and alleviates neuronal damage, muscle damage, neuromuscular junction damage among other aspects. The findings indicate that BI1 can inhibit pathological TDP43 morphology and initially stimulate autophagy through interaction with TDP43. This study establishes a solid theoretical foundation for understanding the regulation of autophagy by BI1 and TDP43 while shedding light on the pathogenesis of ALS through their interaction - offering new concepts and targets for clinical implementation and drug development.

Targeting autoimmune mechanisms by precision medicine in Myasthenia Gravis.

Myasthenia Gravis (MG) is a chronic disabling autoimmune disease caused by autoantibodies to the neuromuscular junction (NMJ), characterized clinically by fluctuating weakness and early fatigability of ocular, skeletal and bulbar muscles. Despite being commonly considered a prototypic autoimmune disorder, MG is a complex and heterogeneous condition, presenting with variable clinical phenotypes, likely due to distinct pathophysiological settings related with different immunoreactivities, symptoms' distribution, disease severity, age at onset, thymic histopathology and response to therapies. Current treatment of MG based on international consensus guidelines allows to effectively control symptoms, but most patients do not reach complete stable remission and require life-long immunosuppressive (IS) therapies. Moreover, a proportion of them is refractory to conventional IS treatment, highlighting the need for more specific and tailored strategies. Precision medicine is a new frontier of medicine that promises to greatly increase therapeutic success in several diseases, including autoimmune conditions. In MG, B cell activation, antibody recycling and NMJ damage by the complement system are crucial mechanisms, and their targeting by innovative biological drugs has been proven to be effective and safe in clinical trials. The switch from conventional IS to novel precision medicine approaches based on these drugs could prospectively and significantly improve MG care. In this review, we provide an overview of key immunopathogenetic processes underlying MG, and discuss on emerging biological drugs targeting them. We also discuss on future direction of research to address the need for patients' stratification in endotypes according with genetic and molecular biomarkers for successful clinical decision making within precision medicine workflow.

Role of complement in myasthenia gravis.

Myasthenia gravis is a prototypic neuroimmune disorder with autoantibodies targeting the acetylcholine receptor complex at the neuromuscular junction. Patients present with mainly ocular muscle weakness and tend to have a generalized muscle weakness later in the clinical course. The weakness can be severe and fatal when bulbar muscles are heavily involved. Acetylcholine receptor antibodies are present in the majority of patients and are of IgG1 and IgG3 subtypes which can activate the complement system. The complement involvement plays a major role in the neuromuscular junction damage and the supporting evidence in the literature is described in this article. Complement therapies were initially studied and approved for paroxysmal nocturnal hemoglobinuria and in the past decade, those have also been studied in myasthenia gravis. The currently available randomized control trial and real-world data on the efficacy and safety of the approved and investigational complement therapies are summarized in this review.

Assay Development and Measurement of Autoantibody-Mediated Complement Activity in Myasthenia Gravis

Autoantibodies against the acetylcholine receptor (AChR) play a critical role in myasthenia gravis, where autoantibody-mediated complement activation has been implicated in neuromuscular junction damage. However, the exact pathogenic role of the autoantibodies in complement activation remains unclear. We developed a cell-based assay that measures AChR autoantibody–mediated complement membrane attack complex (MAC) formation. A modified HEK293T cell line using CRISPR/Cas9 genome editing to disrupt expression of the complement regulator genes (CD46, CD55, and CD59)—was used to measure AChR autoantibody–mediated membrane attack complex (MAC) formation through flow cytometry. We observed a modest correlation between autoantibody-mediated complement mediated activity and disease burden suggesting heterogeneity in autoantibody-mediated activation of complement system.

Serum pentraxin 3 concentration correlates with disease severity in patients with myasthenia gravis

ObjectiveMyasthenia gravis (MG) is an antibody-mediated inflammatory disease affecting post-synaptic membranes of neuromuscular junctions, and objective biomarkers of MG disease activity are lacking. Pentraxin 3 (PTX3) is an acute-phase inflammatory glycoprotein in the same family as C-reactive protein that is associated with disease activity in several autoimmune disorders. Thus, we investigated whether circulating PTX3 is a useful biomarker of MG activity. MethodsSerum PTX3 was measured in 40 patients with MG who were positive for anti-acetylcholine receptor antibody, and in 30 healthy and disease controls, using a commercial enzyme-linked immunosorbent assay kit. In patients with MG, the correlation of serum PTX3 levels with disease severity scales at serum sampling, including MG Foundation of America (MGFA) classification, MG activity of daily living (MG-ADL) score, and quantitative MG (QMG) score, were investigated. ResultsAlthough there was no significant difference in serum PTX3 between the MG and control groups (mean, 3346 pg/mL in MG group vs. 2870 pg/mL in control group, P = 0.56), serum PTX3 moderately correlated with all disease severity scores (MGFA classification: Spearman’s ρ = 0.53, P = 0.0004; MG-ADL score: Spearman’s ρ = 0.45, P = 0.004; QMG score: Spearman’s ρ = 0.50, P = 0.004). ConclusionOur results suggest that circulating PTX3 may reflect the extent of neuromuscular junction damage and might be involved in the pathogenesis of MG.

An electroencephalography-based human-machine interface combined with contralateral C7 transfer in the treatment of brachial plexus injury.

Transferring the contralateral C7 nerve root to the median or radial nerve has become an important means of repairing brachial plexus nerve injury. However, outcomes have been disappointing. Electroencephalography (EEG)-based human-machine interfaces have achieved promising results in promoting neurological recovery by controlling a distal exoskeleton to perform functional limb exercises early after nerve injury, which maintains target muscle activity and promotes the neurological rehabilitation effect. This review summarizes the progress of research in EEG-based human-machine interface combined with contralateral C7 transfer repair of brachial plexus nerve injury. Nerve transfer may result in loss of nerve function in the donor area, so only nerves with minimal impact on the donor area, such as the C7 nerve, should be selected as the donor. Single tendon transfer does not fully restore optimal joint function, so multiple functions often need to be reestablished simultaneously. Compared with traditional manual rehabilitation, EEG-based human-machine interfaces have the potential to maximize patient initiative and promote nerve regeneration and cortical remodeling, which facilitates neurological recovery. In the early stages of brachial plexus injury treatment, the use of an EEG-based human-machine interface combined with contralateral C7 transfer can facilitate postoperative neurological recovery by making full use of the brain’s computational capabilities and actively controlling functional exercise with the aid of external machinery. It can also prevent disuse atrophy of muscles and target organs and maintain neuromuscular junction effectiveness. Promoting cortical remodeling is also particularly important for neurological recovery after contralateral C7 transfer. Future studies are needed to investigate the mechanism by which early movement delays neuromuscular junction damage and promotes cortical remodeling. Understanding this mechanism should help guide the development of neurological rehabilitation strategies for patients with brachial plexus injury.

Nonuniform loss of muscle strength and atrophy during bed rest: a systematic review.

Muscle atrophy and decline in muscle strength appear very rapidly with prolonged disuse or mechanical unloading after acute hospitalization or experimental bed rest. The current study analyzed data from short-, medium-, and long-term bed rest (5-120 days) in a pooled sample of 318 healthy adults and modeled the mathematical relationship between muscle strength decline and atrophy. The results show a logarithmic disuse-induced loss of strength and muscle atrophy of the weight-bearing knee extensor muscles. The greatest rate of muscle strength decline and atrophy occurred in the earliest stages of bed rest, plateauing later, and likely contributed to the rapid neuromuscular loss of function in the early period. In addition, during the first 2 wk of bed rest, muscle strength decline is much faster than muscle atrophy: on day 5, the ratio of muscle atrophy to strength decline as a function of bed rest duration is 4.2, falls to 2.4 on day 14, and stabilizes to a value of 1.9 after ∼35 days of bed rest. Positive regression revealed that ∼79% of the muscle strength loss may be explained by muscle atrophy, while the remaining is most likely due to alterations in single fiber mechanical properties, excitation-contraction coupling, fiber architecture, tendon stiffness, muscle denervation, neuromuscular junction damage, and supraspinal changes. Future studies should focus on neural factors as well as muscular factors independent of atrophy (single fiber excitability and mechanical properties, architectural factors) and on the role of extracellular matrix changes. Bed rest results in nonuniform loss of isometric muscle strength and atrophy over time, where the magnitude of change was greater for muscle strength than for atrophy. Future research should focus on the loss of muscle function and the underlying mechanisms, which will aid in the development of countermeasures to mitigate or prevent the decline in neuromuscular efficiency.NEW & NOTEWORTHY Our study contributes to the characterization of muscle loss and weakness processes reflected by a logarithmic decline in muscle strength induced by chronic bed rest. Acute short-term hospitalization (≤5 days) associated with periods of disuse/immobilization/prolonged time in the supine position in the hospital bed is sufficient to significantly decrease muscle mass and size and induce functional changes related to weakness in maximal muscle strength. By bringing together integrated evaluation of muscle structure and function, this work identifies that 79% of the loss in muscle strength can be explained by muscle atrophy, leaving 21% of the functional loss unexplained. The outcomes of this study should be considered in the development of daily countermeasures for preserving neuromuscular integrity as well as preconditioning interventions to be implemented before clinical bed rest or chronic gravitational unloading (e.g., spaceflights).

Severe worsening of myasthenic symptoms after the eculizumab discontinuation

Myasthenia gravis (MG) is an autoantibody-mediated disease of the neuromuscular junction. The neuromuscular junction damage associated with MG is caused by anti-acetylcholine receptor (AChR) antibody and complements. Recently, eculizumab (an anti-C5 monoclonal antibody) was approved for patients with anti-AChR antibody-positive generalized refractory MG. Here, we report a Japanese man with MG who well responded to eculizumab, but experienced acute severe worsening of myasthenic symptoms 2months after its discontinuation. Plasmapheresis did not improve his symptoms; hence, eculizumab was re-administered, resulting in a dramatic response within a week. This is an informative case because eculizumab discontinuation in patients with MG has been very rarely reported. If eculizumab treatment is clinically well effective and AChR antibody titer does not decrease, clinicians should be aware that acute and critical deterioration of MG may occur after the eculizumab discontinuation.

Identification of biomarkers for physical frailty and sarcopenia through a new multi-marker approach: results from the BIOSPHERE study.

Physical frailty and sarcopenia (PF&S) is a prototypical geriatric condition characterized by reduced physical function and low muscle mass. The aim of the present study was to provide an initial selection of biomarkers for PF&S using a novel multivariate analytic strategy. Two-hundred community-dwellers, 100 with PF&S and 100 non-physically frail, non-sarcopenic (nonPF&S) controls aged 70 and older were enrolled as part of the BIOmarkers associated with Sarcopenia and Physical frailty in EldeRly pErsons (BIOSPHERE) study. A panel of 74 serum analytes involved in inflammation, muscle growth and remodeling, neuromuscular junction damage, and amino acid metabolism was assayed. Biomarker selection was accomplished through sequential and orthogonalized covariance selection (SO-CovSel) analysis. Separate SO-CovSel models were constructed for the whole study population and for the two genders. The model with the best prediction ability obtained with the smallest number of variables was built using seven biomolecules. This model allowed correct classification of 80.6 ± 5.3% PF&S participants and 79.9 ± 5.1% nonPF&S controls. The PF&S biomarker profile was characterized by higher serum levels of asparagine, aspartic acid, and citrulline. Higher serum concentrations of platelet-derived growth factor BB, heat shock protein 72 (Hsp72), myeloperoxidase, and α-aminobutyric acid defined the profile of nonPF&S participants. Gender-specific SO-CovSel models identified a "core" biomarker profile of PF&S, characterized by higher serum levels of aspartic acid and Hsp72 and lower concentrations of macrophage inflammatory protein 1β, with peculiar signatures in men and women.SO-CovSel analysis allowed identifying a set of potential biomarkers for PF&S. The adoption of such an innovative multivariate approach could help address the complex pathophysiology of PF&S, translate biomarker discovery from bench to bedside, and unveil novel targets for interventions.

Impact of sedentarism due to the COVID-19 home confinement on neuromuscular, cardiovascular and metabolic health: Physiological and pathophysiological implications and recommendations for physical and nutritional countermeasures

The COVID-19 pandemic is an unprecedented health crisis as entire populations have been asked to self-isolate and live in home-confinement for several weeks to months, which in itself represents a physiological challenge with significant health risks. This paper describes the impact of sedentarism on the human body at the level of the muscular, cardiovascular, metabolic, endocrine and nervous systems and is based on evidence from several models of inactivity, including bed rest, unilateral limb suspension, and step-reduction. Data form these studies show that muscle wasting occurs rapidly, being detectable within two days of inactivity. This loss of muscle mass is associated with fibre denervation, neuromuscular junction damage and upregulation of protein breakdown, but is mostly explained by the suppression of muscle protein synthesis. Inactivity also affects glucose homeostasis as just few days of step reduction or bed rest, reduce insulin sensitivity, principally in muscle. Additionally, aerobic capacity is impaired at all levels of the O2 cascade, from the cardiovascular system, including peripheral circulation, to skeletal muscle oxidative function. Positive energy balance during physical inactivity is associated with fat deposition, associated with systemic inflammation and activation of antioxidant defences, exacerbating muscle loss. Importantly, these deleterious effects of inactivity can be diminished by routine exercise practice, but the exercise dose–response relationship is currently unknown. Nevertheless, low to medium-intensity high volume resistive exercise, easily implementable in home-settings, will have positive effects, particularly if combined with a 15–25% reduction in daily energy intake. This combined regimen seems ideal for preserving neuromuscular, metabolic and cardiovascular health.

Early Biomarkers of Muscle Atrophy and of Neuromuscular Alterations During 10‐Day Bed Rest

IntroductionDisuse atrophy may not only arise from a reduction in mechanical loading but also from muscle denervation and neuromuscular junction damage, triggered by inactivity [1, 2]. The identification of biomarkers of muscle atrophy and of neuromuscular degeneration is therefore needed for an early detection of neuromuscular alterations induced by inactivity. The aim of this study was to investigate the onset of muscle atrophy and of neuromuscular alterations during a short‐term inactivity period.MethodsTen healthy males (aged 23±5 years) participated to a 10‐day bed rest (BR) study, after Ethical approval and informed consent. Blood samples were collected every two days for the assessment of NMJ damage from serum levels of c‐terminal agrin fragment (CAF). Muscle fibre denervation was determined from the expression of neural cell adhesive molecule (N‐CAM) in myofibres obtained from vastus lateralis (VL) muscle biopsies at baseline, 5 and 10 days of BR. Myofibre atrophy was assessed from mean fibre cross‐sectional area (CSA). Whole muscle atrophy was assessed from measurements of muscle architecture (pennation angle and muscle CSA) every two days of BR using ultrasonography. Significance of differences was tested with repeated‐measures Anova, level of significance was set at p<0.05.ResultsAtrophy of whole muscle and of myofibers, represented by decreases of 5.8% in muscle CSA (P<0.0001), 7.3% in pennation angle (P<0.0001), and by a 21% reduction in mean fibre CSA (P<0.01) were detected after 10 days of BR. A 19% increase (p<0.01) in CAF levels was found on BR day 10, together with a 18‐fold increase in N‐CAM positive myofibres. The analysis of myofiber CSA and N‐CAM expression at BR 5 day is in progress.ConclusionsThese findings show a very early onset (within 10 days) of whole muscle and fiber atrophy accompanied by NMJ damage and myofiber denervation in response to chronic inactivity. We suggest that countermeasures against neuromuscular maladaptation to chronic inactivity ought to be implemented as early as possible.Support or Funding InformationFunding by ASI, MARS‐PRE Project, n. DC‐VUM‐2017‐006, is acknowledged.

Identification of Predictors of Physical Frailty and Sarcopenia through a New Multi‐ Marker Approach

Background and ObjectivesPhysical frailty & sarcopenia (PF&S) is a prototypical geriatric condition characterized by reduced physical function and low muscle mass. Biomarker discovery for PF&S requires a comprehensive approach to cope with the multifaceted pathophysiology of the condition. The aim of the present study was to provide an initial selection of biomarkers for PF&S using a novel multivariate analytic strategy.MethodsTwo‐hundred older community‐dwellers, 100 with PF&S and 100 non‐sarcopenic, non‐frail (nonPF&S) controls aged 70 and older were enrolled as part of the BIOmarkers associated with Sarcopenia and Physical frailty in EldeRly pErsons (BIOSPHERE) study. A panel of 74 circulating analytes involved in inflammation, muscle growth and remodeling, neuromuscular junction damage, and amino acid metabolism was assayed. Biomarker selection was accomplished through sequential and orthogonalized covariance selection (SO‐CovSel) analysis.ResultsSeveral SO‐CovSel models were constructed. The model with the best prediction ability obtained with the smallest number of variables was built using seven biomolecules. This model allowed correct classification of 80.6 ± 5.3% PF&S participants and 79.9 ± 5.1% nonPF&S controls. The PF&S biomarker profile was characterized by higher serum levels of asparagine, aspartic acid, and citrulline. Higher serum concentrations of platelet‐derived growth factor‐BB, heat shock protein 72, myeloperoxidase, and alpha‐aminobutyric acid defined the serum profile of nonPF&S participants.ConclusionSO‐CovSel analysis allowed identifying a set of potential biomarkers for PF&S. The adoption of such an innovative multivariate approach could help address the complex pathophysiology of PF&S, translate biomarker discovery from bench to bedside, and unveil novel targets for interventions.Support or Funding Information‐ Innovative Medicines Initiative – Joint Undertaking (IMI‐JU 115621)‐ Fondazione Roma (NCDs Call for Proposals 2013)‐ Nonprofit research foundation “Centro Studi Achille e Linda Lorenzon”Candidate biomarkers for physical frailty & sarcopenia (PF&S) identified through sequential and orthogonalized covariance selection (SOCovSel) analysis and their putative roles in PF&S pathophysiology. Abbreviations: Hsp72, heat shock protein 72; MPO, myeloperoxidase; PDGF‐BB, platelet‐derived growth factor BB.Figure 1

The disassembly of the neuromuscular synapse in high-fat diet-induced obese male mice

ObjectiveA sustained high fat diet in mice mimics many features of human obesity. We used male and female Non-Swiss albino mice to investigate the impact of short and long-term high-fat diet-(HFD)-induced obesity on the peripheral neuromuscular junction (NMJ) and whether obesity-related synaptic structural alterations were reversible after switching obese mice from HFD to a standard fat diet (SD). MethodsHFD-induced obese and age-matched control mice fed SD were used. We carried out in vivo time lapse imaging to monitor changes of synapses over time, quantitative fluorescence imaging to study the regulation of acetylcholine receptor number and density at neuromuscular junctions, and high resolution confocal microscope to study structural alterations in both the pre- and postsynaptic apparatus. ResultsTime-lapse imaging in vivo over a 9 month period revealed that NMJs of HFD obese male mice display a variety of obesity-related structural alterations, including the disappearance of large synaptic areas, significant reduction in the density/number of nicotinic acetylcholine receptor (AChRs), abnormal distribution of AChRs, high turnover rate of AChRs, retraction of axons from lost postsynaptic sites, and partially denervated synapses. The severity of these synaptic alterations is associated with the duration of obesity. However, no substantial alterations were observed at NMJs of age-matched HFD obese female mice or male mice fed with a standard or low fat diet. Intriguingly, when obese male mice were switched from HFD to a standard diet, receptor density and the abnormal pattern of AChR distribution were completely reversed to normal, whereas lost synaptic structures were not restored. ConclusionsThese results show that the obese male mice are more vulnerable than female mice to the impacts of long-term HFD on the NMJ damage and provide evidence that diet restriction can partially reverse obesity-related synaptic changes.

Ca2+-activated K+ channels modulate microglia affecting motor neuron survival in hSOD1G93A mice

Recent studies described a critical role for microglia in amyotrophic lateral sclerosis (ALS), where these CNS-resident immune cells participate in the establishment of an inflammatory microenvironment that contributes to motor neuron degeneration. Understanding the mechanisms leading to microglia activation in ALS could help to identify specific molecular pathways which could be targeted to reduce or delay motor neuron degeneration and muscle paralysis in patients. The intermediate-conductance calcium-activated potassium channel KCa3.1 has been reported to modulate the “pro-inflammatory” phenotype of microglia in different pathological conditions. We here investigated the effects of blocking KCa3.1 activity in the hSOD1G93AALS mouse model, which recapitulates many features of the human disease. We report that treatment of hSOD1G93A mice with a selective KCa3.1 inhibitor, 1-[(2-chlorophenyl)diphenylmethyl]-1H-pyrazole (TRAM-34), attenuates the “pro-inflammatory” phenotype of microglia in the spinal cord, reduces motor neuron death, delays onset of muscle weakness, and increases survival. Specifically, inhibition of KCa3.1 channels slowed muscle denervation, decreased the expression of the fetal acetylcholine receptor γ subunit and reduced neuromuscular junction damage. Taken together, these results demonstrate a key role for KCa3.1 in driving a pro-inflammatory microglia phenotype in ALS.

Myasthenia gravis: the role of complement at the neuromuscular junction

Generalized myasthenia gravis (gMG) is a rare autoimmune disorder characterized by skeletal muscle weakness caused by disrupted neurotransmission at the neuromuscular junction (NMJ). Approximately 74-88% of patients with gMG have acetylcholine receptor (AChR) autoantibodies. Complement plays an important role in innate and antibody-mediated immunity, and activation and amplification of complement results in the formation of membrane attack complexes (MACs), lipophilic proteins that damage cell membranes. The role of complement in gMG has been demonstrated in animal models and patients. Studies in animals lacking specific complement proteins have confirmed that MAC formation is required to induce experimental autoimmune MG (EAMG) and NMJ damage. Complement inhibition in EAMG models can prevent disease induction and reverse its progression. Patients with anti-AChR+ MG have autoantibodies and MACs present at NMJs. Damaged NMJs are associated with more severe disease, fewer AChRs, and MACs in synaptic debris. Current MG therapies do not target complement directly. Eculizumab is a humanized monoclonal antibody that inhibits cleavage of complement protein C5, preventing MAC formation. Eculizumab treatment improved symptoms compared with placebo in a phase II study in patients with refractory gMG. Direct complement inhibition could preserve NMJ physiology and muscle function in patients with anti-AChR+ gMG.

Cardiovascular effects as a result of a physical conditioning protocol on a critical care inpatient, at three medical centers in Bogotá – Colombia

El síndrome de desacondicionamiento físico es una complicación frecuente en los pacientes con reposo prolongado; su principal característica es la atroia muscular que afecta principalmente las ibras musculares tipo II y se asocia a un daño de la placa neuromuscular. Esto conlleva a múltiples alteraciones metabólicas y sistémicas, como la diicultad para abandonar la ventilación mecánica, la fatiga, el retorno tardío a las actividades de la vida cotidiana y períodos de rehabilitación prolongados. El presente estudio tuvo como objetivo describir los cambios cardiovasculares en pacientes hospitalizados en unidades de cuidado intensivo, al aplicar un protocolo de actividad física dirigida como parte de la rehabilitación. Se trata de un estudio descriptivo tipo serie de casos que se realizó en tres unidades de cuidado intensivo de tipo polivalente durante un año con una muestra de 23 pacientes que tuvieron una estancia superior a 72 horas y que como parte del manejo recibieron ventilación mecánica. Se evaluaron como parámetros cardiovasculares la presión arterial y la frecuencia cardíaca antes, durante y después de cada una de las cargas aplicadas, las cuales fueron ajustadas por un especialista en medicina de la actividad física y del deporte de acuerdo a la evolución clínica y posteriormente registradas en una escala del uno al treinta y dos. Se analizaron los datos de las variables cardiovasculares por medio de estadísticas no paramétricas sin encontrar diferencias signiicativas concluyendo que en pacientes críticos, la utilización de cargas orientadas por parte del especialista idóneo, no tiene efecto cardiovascular considerable y puede mantenerse como parte de los protocolos de rehabilitación. MÉD.UIS. 2016;29(2):161-73.

Response of the Neuromuscular Unit to Spaceflight

Despite the inherent limitations placed on spaceflight investigations, much has been learned about the adaptations of the neuromuscular system to weightlessness from studies of rats flown for relatively short periods (approximately 4-22 days). Below is a summary of the major effects of spaceflight observed in muscles of rats that are not in their rapid growth stage: 1. Skeletal muscles atrophy rapidly during spaceflight; significant atrophy is observed as early as after 4 days of flight. 2. The atrophic response appears to be related to the primary function of the muscle. In the hindlimb, the relative amount of atrophy can be characterized as slow extensors > fast extensors > fast flexors. This pattern of relative atrophy does not appear to be occurring in the forelimb; however, not enough data are available to draw any definitive conclusions at this time. 3. Both slow and fast fibers atrophy during spaceflight, with the largest fibers within an individual muscle generally showing the greatest atrophic response. Interestingly, the amount of fiber atrophy appears to reach a plateau after about 14 days of flight. 4. Adaptations have been observed in the concentration and content of all muscle proteins pools, with the protein pools in slow muscles the most affected. 5. Some slow and fast fibers in predominantly slow and fast muscles show shifts in their histochemical and biochemical properties, toward those observed in a "faster" phenotype. 6. Some fibers, presumably expressing slow MHC isoforms before flight, begin to express fast MHC isoforms during flight. 7. The oxidative capacity of the muscles or fibers is relatively unaffected by spaceflight, particularly in the slow muscles. Any change in whole-body fatigability associated with spaceflight most likely reflects the loss in muscle and fiber mass. 8. The glycolytic capacity of the muscles and muscle fibers is enhanced after spaceflight. This metabolic adaptation seems to be related to the shift in the contractile proteins towards "faster" isoforms. 9. The vascularity of muscles appears to be maintained after flight, based, at least, on histological observations of capillarity. 10. The force capabilities of the muscles and fibers appear to decrease in parallel with the decreases in size, i.e., the specific tension is not significantly affected after flight. 11. Changes in the speed-related properties of the slow muscles are consistent with the adaptations in the myosin molecule, i.e., the slow muscles and some fibers in the slow muscles become "faster." 12. Some muscle fiber and neuromuscular junction damage has been observed after flight, particularly in the slow muscles. The extent of damage may be related to the amount of time that the muscles are allowed to reload before removal, i.e., in general, shorter intervals result in less fiber damage. 13. Adaptations in the motor (ventral horn) and sensory (spinal ganglia) neurons have been quite variable, but this may be related to the amount of time that the muscles are allowed to reload before removal. Morphological adaptations after relatively short periods of reloading may reflect a decrease in the activation of the neural elements during flight.