Motor Neuron Dysfunction Research Articles

Study of motor asymmetry in ALS indicates an effect of limb dominance on onset and spread of weakness, and an important role for upper motor neurons

In amyotrophic lateral sclerosis (ALS), onset and spread of upper motor neuron (UMN) and lower motor neuron (LMN) dysfunction is typically asymmetric. Our aim was to investigate the relationship between limb dominance and the onset and spread of clinical UMN and LMN dysfunction in ALS. We studied 138 ALS subjects with unilateral and concordant limb dominance, from two tertiary centres. A questionnaire was used to determine the pattern of disease onset and spread. The clinical severity of UMN and LMN signs in each limb was quantified using a validated scoring system. Results showed that onset of weakness was more likely to occur in the dominant upper limb (p = 0.02). In subjects with initial weakness in a non-dominant limb, spread of weakness was more likely to be to the other limb on that side (p = 0.008). The relative distribution of upper limb UMN signs was affected by whether weakness first occurred on the dominant or non-dominant side (p = 0.03). These findings support limb dominance as a significant factor underlying onset and spread of ALS, with UMN processes playing an important role. The effect of limb dominance on the presentation of ALS may reflect underlying neuronal vulnerabilities, which become exposed by the disease.

Age‐dependent deterioration of locomotion in Drosophila melanogaster deficient in the homologue of amyotrophic lateral sclerosis 2

Recessive mutations in the amyotrophic lateral sclerosis 2 (ALS2) gene have been linked to juvenile-onset ALS2. Although one of the molecular functions of the ALS2 protein is clearly the activation of Rab5, the mechanisms underlying the selective dysfunction and degeneration of motor neurons in vivo remain to be fully understood. Here, we focused on the ALS2 homologue of Drosophila melanogaster, isolated two independent deletions, and systematically compared phenotypes of the mutants with those of animals in which Rab5 function in identified neurons was abrogated. In the dALS2 mutant flies, we found that the stereotypic axonal and dendritic morphologies of neurons shared some features with those in Rab5-deficient flies, but the dALS2 mutant phenotypes were much milder. We also found that the abrogation of Rab5 function in motor neurons strongly depressed the locomotion activity of adults, resembling the behavior of aged dALS2 mutants. Importantly, this age-dependent locomotion deficit of dALS2 mutants was restored to normal by expressing the dALS2 transgene in a wide range of tissues. This finding provided a platform where we could potentially identify particular cell types responsible for the phenotype by tissue-specific rescue experiments. We discuss our results and the future usage of the dALS2 mutant as a new ALS model.

Diagnostic utility of cortical excitability studies in amyotrophic lateral sclerosis

The diagnosis of amyotrophic lateral sclerosis (ALS) relies on identification of a combination of upper and lower motor neuron signs. In order to improve the diagnostic sensitivity for ALS, Awaji criteria were developed, in part to better incorporate neurophysiological measures, although assessment of upper motor neuron dysfunction remained clinically based. Given that cortical hyperexcitability appears to be an early feature in ALS, the present study assessed the diagnostic utility of a threshold tracking transcranial magnetic stimulation technique as an aid to the research-based Awaji criteria in establishing an earlier diagnosis of ALS. Prospective studies were undertaken on a cohort of 82 patients with suspected ALS and results were compared with 34 healthy controls. Short-interval intracortical inhibition (SICI) was significantly reduced in ALS patients (P<0.0001), with a comparable reduction evident in the Awaji groups (SICIAWAJI POSSIBLE 1.3%±1.3%; SICIAWAJI PROBABLE/DEFINITE 1.4%±1.7%). Central motor conduction time was significantly prolonged (P<0.001), whereas the motor evoked potential amplitude (P<0.05) and intracortical facilitation (P<0.05) were increased. The frequency of transcranial magnetic stimulation abnormalities was similar across Awaji subgroups, and addition of transcranial magnetic stimulation abnormalities as a diagnostic category enabled reclassification of 88% of Awaji possible patients to Awaji probable/definite. Cortical excitability studies potentially facilitate an earlier diagnosis of ALS when combined with clinical and conventional neurophysiological findings, albeit in a research setting.

Blood–spinal cord barrier disruption contributes to early motor-neuron degeneration in ALS-model mice

Humans with ALS and transgenic rodents expressing ALS-associated superoxide dismutase (SOD1) mutations develop spontaneous blood-spinal cord barrier (BSCB) breakdown, causing microvascular spinal-cord lesions. The role of BSCB breakdown in ALS disease pathogenesis in humans and mice remains, however, unclear, although chronic blood-brain barrier opening has been shown to facilitate accumulation of toxic blood-derived products in the central nervous system, resulting in secondary neurodegenerative changes. By repairing the BSCB and/or removing the BSCB-derived injurious stimuli, we now identify that accumulation of blood-derived neurotoxic hemoglobin and iron in the spinal cord leads to early motor-neuron degeneration in SOD1(G93A) mice at least in part through iron-dependent oxidant stress. Using spontaneous or warfarin-accelerated microvascular lesions, motor-neuron dysfunction and injury were found to be proportional to the degree of BSCB disruption at early disease stages in SOD1(G93A) mice. Early treatment with an activated protein C analog restored BSCB integrity that developed from spontaneous or warfarin-accelerated microvascular lesions in SOD1(G93A) mice and eliminated neurotoxic hemoglobin and iron deposits. Restoration of BSCB integrity delayed onset of motor-neuron impairment and degeneration. Early chelation of blood-derived iron and antioxidant treatment mitigated early motor-neuronal injury. Our data suggest that BSCB breakdown contributes to early motor-neuron degeneration in ALS mice and that restoring BSCB integrity during an early disease phase retards the disease process.

15. Does the selective membrane potential abnormality precede the onset of the ALS?

Introduction In ALS patients, peripheral ion channel abnormality and membrane potential changes often exist. A recent study suggests selective membrane depolarization in some ALS patients. We performed the threshold tracking test in patient with upper motor neuron dysfunction. Patients Patient 1 is a 52-year-old man, whose chief complaint was gait disturbance for 4 years. He had no muscle weakness or fasciculations. Deep tendon reflexes (DTR) were exaggerated. The median CMAP amplitude was decreased (2.89 mV). EMG was normal except for multiplet of the quadriceps femoris. Patient 2 is a 69-year-old woman, whose chief complaint was dysarthria for 2 years. She had no muscle weakness or fasciculations. DTRs were exaggerated and jaw reflex was positive. Needle EMG showed chronic denervation only in the tongue. Result Two patients did not fulfill the criteria of ALS. We did threshold tracking in the median motor nerve. Target threshold CMAP amplitudes were set at 10%, 20%, 40% and 60% of the maximum CMAP. Both patients showed normal TE (threshold electrotonus) at 40%. The threshold changes by the hyperpolarizing current was small at the low target levels that suggests membrane depolarization. Conclusion Selective membrane depolarization may precede overt clinical and routine electrophysiological findings.

Are we really closer to improving the diagnostic sensitivity in ALS patients with Awaji criteria?

The Awaji criteria, recently introduced to increase diagnosis sensitivity in amyotrophic lateral sclerosis (ALS), equate the diagnostic significance of neurogenic electrophysiological changes to clinical signs of lower motor neuron dysfunction. They also increase the electrophysiological significance of fasciculation potentials (FPs). The aim of our study was to analyse whether the new parameters improve diagnostic sensitivity in ALS patients primarily diagnosed with the El Escorial criteria. Medical and electrophysiological records of 135 consecutive patients with ALS and 25 patients with progressive muscular atrophy (PMA) who underwent electrophysiological examination of at least three anatomical regions were analysed retrospectively. Results showed that implementation of the Awaji criteria increased the level of ALS diagnosis sensitivity in 5.9% of cases – 1.5% due to the new role of FPs potentials and 4.4% because of equalization of clinical and EMG findings. In 4% of patients the ALS diagnosis was, however, changed from laboratory-supported probable ALS to possible ALS. In conclusion, our study confirms that Awaji modifications are able to improve the diagnostic certainty in a few ALS cases. Although the new approach to FPs markedly increases the number of involved muscles, it only slightly raises the number of involved regions.

Distinct patterns of cortical atrophy in ALS patients with or without dementia: An MRI VBM study

Voxel based morphometry (VBM) allows objective and automated detection of structural changes in brains of patients with amyotrophic lateral sclerosis (ALS). We investigated whether VBM could identify cortical atrophy from T1-weighted images obtained during routine 1.5T studies of ALS patients with various clinically defined phenotypes. For this purpose T1-weighted brain MRI was obtained at 1.5T during routine clinical study in neurologic disease controls (n = 15) and ALS patients (n = 88) categorized into four subgroups based on their clinical phenotypes: predominant upper motor neuron (UMN) dysfunction with or without corticospinal tract (CST) hyperintensity (ALS-CST+/–), combined UMN and prominent lower motor neuron (LMN) dysfunction (classic ALS), and frontotemporal dementia (ALS-FTD). VBM analysis of gray matter (GM) was carried out using FSL. Results demonstrated that clinically obtained brain MRI at 1.5T revealed significantly reduced GM volume in brains of only ALS-FTD patients and not of those with predominant UMN dysfunction or classic ALS, compared to neurologic disease controls. In conclusion, GM volume loss in motor and extramotor regions of only ALS patients with FTD and not of ALS patients without FTD suggests distinct sites of predominant pathology and possibly of disease onset. Brain volumetric measures supplemented by histopathological correlations and other neuroimaging techniques, such as diffusion tensor imaging, may provide insight into ALS pathophysiology.

Maternal MDMA administration in mice leads to neonatal growth delay

The psychoactive recreational drug 3,4-methylenedioxymethamphetamine (MDMA) is widely abused. The fact that MDMA induces neurotoxic damage in serotonergic nerve endings is well known. However, the effects of MDMA on pregnant and neonatal animals remain unknown. Therefore, we studied the effects of gestational exposure to MDMA on birth, growth, and behavior of pups. Female BALB/c mice were orally administered either water (10 ml/kg) or MDMA (20 mg/10 ml/kg) from gestational day 1 to postnatal day (P) 21. MDMA did not affect the birth rate, but the survival rate of the pups significantly decreased. A significant reduction in body weight gain was observed in pups from MDMA-administered dams during P3-P21. Maternal MDMA treatment caused an attenuated cliff avoidance reaction and decreased motor function in the pups, as determined by the wire hanging test. These results suggest that MDMA treatment during pregnancy and lactation causes growth retardation and dysfunction of motor neurons in mouse pups.

Astrocytes expressing mutant SOD1 and TDP43 trigger motoneuron death that is mediated via sodium channels and nitroxidative stress

Amyotrophic lateral sclerosis (ALS) is a fatal paralytic disorder caused by dysfunction and degeneration of motor neurons. Multiple disease-causing mutations, including in the genes for SOD1 and TDP-43, have been identified in ALS. Astrocytes expressing mutant SOD1 are strongly implicated in the pathogenesis of ALS: we have shown that media conditioned by astrocytes carrying mutant SOD1G93A contains toxic factor(s) that kill motoneurons by activating voltage-sensitive sodium (Nav) channels. In contrast, a recent study suggests that astrocytes expressing mutated TDP43 contribute to ALS pathology, but do so via cell-autonomous processes and lack non-cell-autonomous toxicity. Here we investigate whether astrocytes that express diverse ALS-causing mutations release toxic factor(s) that induce motoneuron death, and if so, whether they do so via a common pathogenic pathway. We exposed primary cultures of wild-type spinal cord cells to conditioned medium derived from astrocytes (ACM) that express SOD1 (ACM-SOD1G93A and ACM-SOD1G86R) or TDP43 (ACM-TDP43A315T) mutants; we show that such exposure rapidly (within 30–60 min) increases dichlorofluorescein (DCF) fluorescence (indicative of nitroxidative stress) and leads to extensive motoneuron-specific death within a few days. Co-application of the diverse ACMs with anti-oxidants Trolox or esculetin (but not with resveratrol) strongly improves motoneuron survival. We also find that co-incubation of the cultures in the ACMs with Nav channel blockers (including mexiletine, spermidine, or riluzole) prevents both intracellular nitroxidative stress and motoneuron death. Together, our data document that two completely unrelated ALS models lead to the death of motoneuron via non-cell-autonomous processes, and show that astrocytes expressing mutations in SOD1 and TDP43 trigger such cell death through a common pathogenic pathway that involves nitroxidative stress, induced at least in part by Nav channel activity.

Differentiation defects in primary motoneurons from a SMARD1 mouse model that are insensitive to treatment with low dose PEGylated IGF1.

Muscle atrophy and diaphragmatic palsy are the clinical characteristics of spinal muscular atrophy with respiratory distress type 1 (SMARD1), and are well represented in the neuromuscular degeneration (Nmd2J) mouse, modeling the juvenile form of SMARD1. Both in humans and mice mutations in the IGHMBP2 gene lead to motoneuron degeneration. We could previously demonstrate that treatment with a polyethylene glycol-coupled variant of IGF1 (PEG-IGF1) improves motor functions accompanied by reduced fiber degeneration in the gastrocnemius muscle and the diaphragm, but has no beneficial effect on motoneuron survival. These data raised the question which cell autonomous disease mechanisms contribute to dysfunction and loss of Ighmbp2-deficient motoneurons. An analysis of primary Ighmbp2-deficient motoneurons exhibited differentiation deficits such as reduced spontaneous Ca2+ transients and altered axon elongation, which was not compensated by PEG-IGF1. This points to an IGF1 independent mechanism of motoneuron degeneration that deserves treatment approaches in addition to IGF1.

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Introduction: 79 year old male presented with generalized weakness, slurred speech, altered mental status and labored breathing for several hours along with 20 pound weight loss over 6 months. Patient had three admissions at different hospital over the last 3 months for similar but milder presentation. Past work up included CT scan of the chest which was unrevealing and pulmonary function test (PFT) which showed restrictive lung disease with FEV1 of 48%, FVC of 44% of predicted and FEV1/FVC 84, though he was given the diagnosis of chronic obstructive pulmonary disease (COPD) as an explanation of his hyperbaric respiratory failure. Physical exam in the ED was pertinent for tachycardia, labored shallow breathing and lethargy. Auscultation of his chest demonstrated bilateral decreased breath sound. An arterial blood gas analysis showed pH of 7.04, PaCO2 of >130.00 mmHg and PaO2 of 218 mmHg on 100% oxygen. Chest x-ray and CT scan of the head were both unremarkable for acute process. Hypercarbia was initially attributed to possible COPD and obstructive sleep apnea (OSA). Patient's trachea immediately intubated and mechanical ventilation was begun. After borderline spontaneous breathing trial, patient was extubated next morning. Patient required bi-level noninvasive positive pressure mechanical ventilation (NIPPV) for moderate respiratory distress after extubation. A repeat physical examination was remarkable for impaired dorsiflexion of right ankle, tongue fasciculation and weak negative inspiratory force. There was no evidence of upper motor neuron dysfunction. Work up for neuromuscular weakness included antiacetylcholine receptor antibody, which was negative and electromyography, which demonstrated 2+ fasciculation and fibrillation potentials in >3 muscle groups. A diagnosis of progressive muscular atrophy (PMA) with early respiratory failure was made; it is believed to be a rare form of Amyotrophic Lateral Sclerosis (ALS). After discussing the merits and demerits of invasive mechanical ventilation via tracheostomy and gastrostomy tube insertion, he opted for comfort care. ALS is a neurodegenerative disease characterized by progressive muscular paralysis due to degeneration of lower and upper motor neurons in the primary motor cortex, brainstem, and spinal cord. Muscular paralysis is progressive and typically leads to death due to respiratory failure within 2-5 years of onset. However, only 3.0% of patients with ALS present with respiratory symptoms and 14% of this subpopulation requires mechanical respiratory support, a rare presentation of ALS. Clinical features of early respiratory failure like dyspnea on minor exertion, early morning headache, daytime fatigue and weight loss may be subtle and often overlooked. The classic form of ALS presents with both upper motor neuron (UMN) and lower motor neuron (LMN) findings, but it can be as in our case limited to LMN only and is referred to as PMA. Compared to ALS, patients with PMA are more likely to be males and live longer. Management of ALS in all of its forms is supportive, multidisciplinary and palliative. Early use of NIPPV in ALS patients prolong survival and improve quality of life. Invasive mechanical ventilation, implying the use of an endotracheal tube or a tracheostomy may be considered. Patient with bulbar dysfunction may need gastrostomy to support nutrition. A FVC of <50% of the predicted value has been shown to be associated with poor prognosis. Arriving to accurate diagnosis in timely manner is critical for health care team to expedited discussion with patient and family about management plan and encourage them to consider advanced directives regarding end of life. Patients should be provided with a realistic prognostication of the course of their disease and in the event of decline of full mechanical ventilation, patient and family should be provided with assurance that palliative care strategies can control symptoms in the terminal phase of illness.

Human superoxide dismutase 1 overexpression in motor neurons of Caenorhabditis elegans causes axon guidance defect and neurodegeneration

Strong evidence indicates that mutant Cu, Zn-superoxide dismutase 1 (SOD1) exerts toxic effect on motor neurons in amyotrophic lateral sclerosis (ALS). However, the nature of mutant SOD1-mediated motor neuron degeneration is poorly understood. To provide new insight into the mechanism by which mutant SOD1 induces motor neuron injury, we developed novel Caenorhabditis elegans models of ALS. Expression of human wild type or G93A SOD1 specifically in motor neurons of C. elegans caused progressive locomotion defect and paralytic phenotype, which recapitulate some characteristic features of ALS including age-dependent motor dysfunction and degeneration of motor neurons associated with SOD1 aggregation. In addition, the motor neuron loss is independent of cell death protein 3 (CED-3)/cell death protein 4 (CED-4) caspase pathway. We also found that before motor neurons began to die in adulthood, axon guidance defect of motor neuron appeared during the development stages. When green fluorescent protein (GFP)-tagged proteins related to axon guidance were examined in motor neurons, a significantly decreased density and number of GFP-tagged puncta were observed in the transgenic worms. Our models mimic axon developmental defect and the adult-onset degeneration of motor neurons in ALS. Using this model, we uncovered the cell-autonomous damage caused by human SOD1 to motor neurons in vivo, and provided a new insight into the developmental defect mechanism that may contribute to motor neuron degeneration in ALS.

Excitation BolsTORs Motor Neurons in ALS Mice

It is unclear why motor neurons selectively degenerate in amyotrophic lateral sclerosis (ALS). Saxena etal. (2013) demonstrate that excitation of motor neurons can prevent their demise in a mouse model of inherited ALS by a mechanism involving the mTOR pathway.

P.17.9 AAV9 vector encoding hGAA improves synaptic pathology of the neuromuscular junction in Pompe disease

Pompe disease is caused by the deficiency of the lysosomal enzyme acid alpha-glucosidase (GAA) and results in a progressive accumulation of lysosomal glycogen. This impacts cellular architecture and function and leads to progressive respiratory insufficiency and motoneuron dysfunction. We have developed an adeno-associated virus (AAV) vector strategy to express long-term therapeutic levels of human GAA in cardiac, skeletal muscle, and the CNS resulting in improved cardiorespiratory function. However, an incomplete restoration of skeletal muscle function or activation still remains. We tested the hypothesis that an AAV9 vector encoding hGAA would result in retrograde transport and restore neuromuscular junction integrity and reverse axonal pathology in Pompe disease. 20 month old Pompe mice (Gaa−/−) were randomized to the following groups: untreated (Gaa−/−), AAV9-CMV-hGAA, or AAV9-DES-hGAA. AAV9 treated animals received a single injection of 1 × 1011vg in the right tibialis anterior (TA) muscle. One month post injection, the TA muscle and lumbar spinal cord were analyzed for vector genome copy number and GAA activity. Significant levels of vector genomes were detected in the TA (AAV9-DES-hGAA 1.5 × 105 ± 3.1 × 104 vg/ug DNA; AAV9-CMV-hGAA 8.4 × 104 ± 1.7 × 104 vg/ug DNA) and lumbar spinal cord (AAV9-DES-hGAA 1.5 × 103 ± 1.7 × 102 vg/ug DNA; AAV9-CMV-hGAA 2.5 × 103 ± 1.3 × 103 vg/ug DNA) suggesting efficient transduction of skeletal muscle and retrograde transport of AAV9. Activity of GAA in TA lysates was 2396% and 1770% above wild-type in AAV9-DES-hGAA and AAV9-CMV-hGAA animals, respectively ( p

Brain white matter diffusion tensor metrics from clinical 1.5T MRI distinguish between ALS phenotypes

Patients with amyotrophic lateral sclerosis (ALS) can present with varying degrees of upper motor neuron (UMN) and lower motor neuron dysfunction. Previous diffusion tensor imaging (DTI) studies, in which ALS patients were not separated by the degree of UMN dysfunction, have resulted in conflicting or inconclusive results. We hypothesized that (1) categorizing ALS patients by their clinical phenotype can reveal differences in DTI abnormalities along the corticospinal tract (CST), and (2) data obtained from routine clinical DTI scans can provide this type of information. Clinical DTI scans were obtained at 1.5T in 87 ALS patients (categorized into four subgroups based on clinical phenotype) and in 12 neurologic controls. Fractional anisotropy (FA), mean diffusivity (MD), axial diffusivity (AD) and radial diffusivity values from the CST were compared between ALS subgroups and controls. Significantly reduced FA and elevated MD values were observed in ALS patients compared to controls at the subcortical motor cortex level. Significant differences in AD values were not only seen between control and ALS patients but also between the ALS subgroups, suggesting divergent pathologies in these ALS patients. Classifying ALS patients by phenotype reveals differences in CST abnormalities between subgroups and may provide novel insights into disease mechanisms. The close similarity of our results from routine clinical scans with published research studies suggests wider accessibility to useful DTI metrics.

Neural Induction with Neurogenin 1 Enhances the Therapeutic Potential of Mesenchymal Stem Cells in an Amyotrophic Lateral Sclerosis Mouse Model

Amyotrophic lateral sclerosis (ALS) is characterized by progressive dysfunction and degeneration of motor neurons in the central nervous system (CNS). In the absence of effective drug treatments for ALS, stem cell treatment has emerged as a candidate therapy for this disease. To date, however, there is no consensus protocol that stipulates stem cell types, transplantation timing, or frequency. Using an ALS mouse model carrying a high copy number of a mutant human superoxide dismutase-1 (SOD1)(G93A) transgene, we investigated the effect of neural induction on the innate therapeutic potential of mesenchymal stem cells (MSCs) in relation to preclinical transplantation parameters. In our study, the expression of monocyte chemoattractant protein-1 (MCP-1) was elevated in the ALS mouse spinal cord. Neural induction of MSCs with neurogenin 1 (Ngn1) upregulated the expression level of the MCP-1 receptor, CCR2, and enhanced the migration activity toward MCP-1 in vitro. Ngn1-expressing MSCs (MSCs-Ngn1) showed a corresponding increase in tropism to the CNS after systemic transplantation in ALS mice. Notably, MSCs-Ngn1 delayed disease onset if transplanted during preonset ages,whereas unprocessed MSCs failed to do so. If transplanted near the onset ages, a single treatment with MSCs-Ngn1 was sufficient to enhance motor functions during the symptomatic period (15–17 weeks), whereas unprocessed MSCs required repeated transplantation to achieve similar levels of motor function improvement. Our data indicate that systemically transplanted MSCs-Ngn1 can migrate to the CNS and exert beneficial effects on host neural cells for an extended period of time through paracrine functions, suggesting a potential benefit of neural induction of transplanted MSCs in long-term treatment of ALS.

Motor neuron involvement in multisystem proteinopathy

To explore the putative connection between inclusion body myopathy, Paget disease, frontotemporal dementia (IBMPFD) and motor neuron disease (MND). Clinical, genetic, and EMG characterization of 17 patients from 8 IBMPFD families. Limb weakness was the most common clinical manifestation (present in 15 patients, median onset age 38 years, range 25-52), with unequivocal evidence of upper motor neuron dysfunction in 3. EMG, abnormal in all 17, was purely neurogenic in 4, purely myopathic in 6, and mixed neurogenic/myopathic in 7. Cognitive/behavioral impairment was detected in at least 8. Mutations in VCP (R155H, R159G, R155C) were identified in 6 families, and in hnRNPA2B1 (D290V) in another family. The genetic cause in the eighth family has not yet been identified. Mutations in at least 4 genes may cause IBMPFD, and its phenotypic spectrum extends beyond IBM, Paget disease, and frontotemporal dementia (FTD). Weakness, the most common and disabling manifestation, may be caused by muscle disease or MND. The acronym IBMPFD is, therefore, insufficient to describe disorders due to VCP mutations or other recently identified IBMPFD-associated genes. Instead, we favor the descriptor multisystem proteinopathy (MSP), which encompasses both the extended clinical phenotype and the previously described prominent pathologic feature of protein aggregation in affected tissues. The nomenclature MSP1, MSP2, and MSP3 may be used for VCP-, HNRNPA2B1-, and HNRNPA1-associated disease, respectively. Genetic defects in MSP implicate a range of biological mechanisms including RNA processing and protein homeostasis, both with potential relevance to the pathobiology of more common MNDs such as amyotrophic lateral sclerosis (ALS) and providing an additional link between ALS and FTD.

Motor neuron dysfunction in a mouse model of ALS: Gender-dependent effect of P2X7 antagonism

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative progressive currently untreatable disease, characterized by selective motor neuron degeneration; the incidence and prevalence of ALS are greater in men than in women. Although some important mechanisms that might contribute to the death of motor neurons have been identified, the mechanisms underlying disease pathophysiology are still uncertain. In particular, the mechanisms underlying the role of gender in ALS and whether treatments should take into account sexual dimorphism remain only partially understood. Recently, the P2X7 receptor for ATP was reported to display neurotoxic potential in motor neuron disorders, and antagonism of the receptor has been suggested to be helpful in these disorders. Studying transgenic mice with superoxide dismutase 1 gene mutations, widely used as model for ALS, may provide a better understanding of pathogenic mechanisms and of toxicity towards motor neurons, also possibly helping to understand whether treatments for ALS should take into account sexual dimorphism.The aim of the work was (1) investigating on gender-dependence of disease progression in the standard model for ALS – the transgenic mouse bearing superoxide dismutase 1 gene mutations – and (2) assessing if a P2X7 receptor antagonist treatment should take into account sexual dimorphism.We evaluated if gender affect the disease course, the motor performance, the weight loss and the lifespan in mice overexpressing mutant superoxide dismutase 1. We measured motor impairment, motor strength and coordination by rotarod and grip strength testing. Further, we assessed if a treatment with the P2X7 receptor antagonist Brilliant Blue G – a dye that can cross the blood–brain barrier, has low toxicity, and has exhibited therapeutic effects in animal models of neurodegenerative diseases – impact on the disease progression, in male and female ALS mice.We found that (1) the onset and the disease progression, and the survival were dependent on gender: male performed worst than female, lost body weight and died before; (2) treatment with the P2X7 receptor antagonist Brilliant Blue G ameliorated the disease progression. The treatment effect was gender-dependent: amelioration was greater in male than in female.In conclusions, we suggest that not only pathogenetic mechanism of motor neuron toxicity but also the drug treatment effectiveness may depend on gender; sexual dimorphism should be considered when investigating on ALS treatment efficacy in the ALS animal model. Our findings also point on the potential relevance of P2X7 receptor antagonism for ALS treatment, and highlight the importance of adopting a sex-specific approach to searching for treatment of ALS.

Mutant SOD1-expressing astrocytes release toxic factors that trigger motoneuron death by inducing hyperexcitability

Amyotrophic lateral sclerosis (ALS) is a devastating paralytic disorder caused by dysfunction and degeneration of motoneurons starting in adulthood. Recent studies using cell or animal models document that astrocytes expressing disease-causing mutations of human superoxide dismutase 1 (hSOD1) contribute to the pathogenesis of ALS by releasing a neurotoxic factor(s). Neither the mechanism by which this neurotoxic factor induces motoneuron death nor its cellular site of action has been elucidated. Here we show that acute exposure of primary wild-type spinal cord cultures to conditioned medium derived from astrocytes expressing mutant SOD1 (ACM-hSOD1(G93A)) increases persistent sodium inward currents (PC(Na)), repetitive firing, and intracellular calcium transients, leading to specific motoneuron death days later. In contrast to TTX, which paradoxically increased twofold the amplitude of calcium transients and killed motoneurons, reduction of hyperexcitability by other specific (mexiletine) and nonspecific (spermidine and riluzole) blockers of voltage-sensitive sodium (Na(v)) channels restored basal calcium transients and prevented motoneuron death induced by ACM-hSOD1(G93A). These findings suggest that riluzole, the only FDA-approved drug with known benefits for ALS patients, acts by inhibiting hyperexcitability. Together, our data document that a critical element mediating the non-cell-autonomous toxicity of ACM-hSOD1(G93A) on motoneurons is increased excitability, an observation with direct implications for therapy of ALS.

Tempol Moderately Extends Survival in a hSOD1G93A ALS Rat Model by Inhibiting Neuronal Cell Loss, Oxidative Damage and Levels of Non-Native hSOD1G93A Forms

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the progressive dysfunction and death of motor neurons by mechanisms that remain unclear. Evidence indicates that oxidative mechanisms contribute to ALS pathology, but classical antioxidants have not performed well in clinical trials. Cyclic nitroxides are an alternative worth exploring because they are multifunctional antioxidants that display low toxicity in vivo. Here, we examine the effects of the cyclic nitroxide tempol (4-hydroxy-2,2,6,6-tetramethyl piperidine-1-oxyl) on ALS onset and progression in transgenic female rats over-expressing the mutant hSOD1G93A . Starting at 7 weeks of age, a high dose of tempol (155 mg/day/rat) in the rat´s drinking water had marginal effects on the disease onset but decelerated disease progression and extended survival by 9 days. In addition, tempol protected spinal cord tissues as monitored by the number of neuronal cells, and the reducing capability and levels of carbonylated proteins and non-native hSOD1 forms in spinal cord homogenates. Intraperitoneal tempol (26 mg/rat, 3 times/week) extended survival by 17 days. This group of rats, however, diverted to a decelerated disease progression. Therefore, it was inconclusive whether the higher protective effect of the lower i.p. dose was due to higher tempol bioavailability, decelerated disease development or both. Collectively, the results show that tempol moderately extends the survival of ALS rats while protecting their cellular and molecular structures against damage. Thus, the results provide proof that cyclic nitroxides are alternatives worth to be further tested in animal models of ALS.