Progressive impairment of CaV1.1 function in the skeletal muscle of mice expressing a mutant type 1 Cu/Zn superoxide dismutase (G93A) linked to amyotrophic lateral sclerosis
© The Author(s). 2016
Received: 1 October 2015
Accepted: 3 June 2016
Published: 23 June 2016
Amyotrophic lateral sclerosis (ALS) is an adult-onset neurodegenerative disorder that is typically fatal within 3–5 years of diagnosis. While motoneuron death is the defining characteristic of ALS, the events that underlie its pathology are not restricted to the nervous system. In this regard, ALS muscle atrophies and weakens significantly before presentation of neurological symptoms. Since the skeletal muscle L-type Ca2+ channel (CaV1.1) is a key regulator of both mass and force, we investigated whether CaV1.1 function is impaired in the muscle of two distinct mouse models carrying an ALS-linked mutation.
We recorded L-type currents, charge movements, and myoplasmic Ca2+ transients from dissociated flexor digitorum brevis (FDB) fibers to assess CaV1.1 function in two mouse models expressing a type 1 Cu/Zn superoxide dismutase mutant (SOD1G93A).
In FDB fibers obtained from “symptomatic” global SOD1G93A mice, we observed a substantial reduction of SR Ca2+ release in response to depolarization relative to fibers harvested from age-matched control mice. L-type current and charge movement were both reduced by ~40 % in symptomatic SOD1G93A fibers when compared to control fibers. Ca2+ transients were not significantly reduced in similar experiments performed with FDB fibers obtained from “early-symptomatic” SOD1G93A mice, but L-type current and charge movement were decreased (~30 and ~20 %, respectively). Reductions in SR Ca2+ release (~35 %), L-type current (~20 %), and charge movement (~15 %) were also observed in fibers obtained from another model where SOD1G93A expression was restricted to skeletal muscle.
We report reductions in EC coupling, L-type current density, and charge movement in FDB fibers obtained from symptomatic global SOD1G93A mice. Experiments performed with FDB fibers obtained from early-symptomatic SOD1G93A and skeletal muscle autonomous MLC/SOD1G93A mice support the idea that events occurring locally in the skeletal muscle contribute to the impairment of CaV1.1 function in ALS muscle independently of innervation status.
Amyotrophic lateral sclerosis (ALS), known commonly as Lou Gehrig’s disease in the USA or motoneurone disease in the UK, is an adult-onset neurodegenerative disorder that is usually lethal within 3 to 5 years of initial diagnosis [1, 2]. As a consequence of disrupted nerve-muscle communication, afflicted individuals progressively lose control of voluntary muscle function and die most frequently from respiratory failure. Nearly a quarter of familial ALS cases have been linked to point mutations in the type 1 Cu/Zn superoxide dismutase (SOD1) enzyme, which functions to mitigate cellular oxidative stress . Since these mutations are autosomal dominant, transgenic mice overexpressing ALS-linked SOD1 mutations have proven to be useful models for investigation of ALS pathogenesis [4, 5].
Two independent groups have found that skeletal muscle-targeted, transgenic overexpression of SOD1G93A causes profound muscle atrophy [6, 7] and initiates motoneuron death . These congruent reports reinforce earlier challenges to the dogma that ALS is solely a neurological disorder and support the “dying-back” phenomenon by which motor unit loss and associated muscle function precede the death of motor neurons (reviewed in refs. [8–11]). Additional support for this view is provided by the observations that destruction of neuromuscular junctions has been linked to oxidative stress induced by tissue-specific breakdown of muscle mitochondria  and trophic factors secreted from the skeletal muscle promote motoneuron survival in an ALS model by stabilizing neuromuscular junctions (e.g., IGF-1, GDNF) [13–15].
In the skeletal muscle, CaV1.1 functions both as the voltage-sensor for excitation-contraction (EC) coupling and as an L-type Ca2+ channel [16–18]. CaV1.1’s role as voltage-sensor is firmly established , but recent work has also revealed that L-type Ca2+ entry maintains myoplasmic Ca2+ levels during repetitive activity [20, 21] augments muscle contraction , engages excitation-transcription coupling , regulates energy expenditure , and promotes development of neuromuscular junctions . Thus, aberrant CaV1.1 activity may profoundly affect skeletal muscle biology.
The question of whether CaV1.1 function is disrupted during the progression of ALS was originally posed over 20 years ago [26–28], but the experimental approach used to investigate this premise left much ambiguity. In these earlier studies, serum of ALS patients was acutely applied to normal cut wild-type rat extensor digitorum longus (EDL) fibers and the Vaseline-gap voltage-clamp technique was used to record L-type currents and gating charge movements [26, 27]; L-type currents and intramembrane gating charge movements were both found to be reduced. Similarly, acute IgG application also produced inhibitory effects on single CaV1.1 channels in planar lipid bilayers . In the time that has passed since the experiments that employed acute application of ALS IgG were performed, it has become evident that that muscle dysfunction in ALS manifests early and slowly over time. Fortunately, a number of ALS mouse models have been developed since then, including SOD1G93A transgenic lines [4, 29–31].
Here, we provide the first comprehensive voltage-clamp assessment of CaV1.1 function in the muscle of an engineered mammalian model of ALS. Specifically, we utilized both global SOD1G93A mice and mice expressing SOD1G93A only in skeletal muscle to systematically investigate whether CaV1.1 function is compromised during the progression of the disease. We found substantial reductions in myoplasmic Ca2+ transient amplitude, peak L-type current density, and maximal intramembrane charge movement in flexor digitorum brevis (FDB) fibers obtained from “symptomatic” congenic SOD1G93A mice. Consistent with a progressive muscle defect, we observed lesser reductions in L-type current amplitude and charge movement in FDB fibers obtained from “early-symptomatic” SOD1G93A mice in which widespread denervation had yet to occur. To investigate the possibility that events occurring in muscle contribute, at least in part, to the impairment of CaV1.1 function, we performed voltage-clamp experiments with FDB fibers of mice expressing SOD1G93A autonomously in skeletal muscle. In these experiments, we observed significant reductions in EC coupling, charge movement, and peak L-type current amplitude.
All procedures involving mice were approved by the University of Colorado-Anschutz Medical Campus Institutional Animal Care and Use Committee (91813(05)1D). Male congenic SOD1G93A mice (B6.Cg-Tg(SOD1*G93A)1Gur/J; 32 transgene copies) and age-matched male C57BL/6J mice (both Jackson Laboratories, Bar Harbor, ME) were used at two time points based on the criteria for disease progression described by Hatzipetros et al. : (1) “symptomatic” and (2) “early-symptomatic.” Nine symptomatic mice presented classical ALS symptoms (i.e., tremor, abnormal gait, comprised ability to stand on hindquarters, etc.) when they were sacrificed at 149 ± 1 days. Six mice from the early-symptomatic cohort were largely devoid of overt symptoms when they were sacrificed at 105 ± 1 days. Age-matched C57BL/6J mice were used as background wild-type controls for both symptomatic and early-symptomatic global SOD1G93A mice (eight and seven mice, respectively). Seven MLC/SOD1G93A mice were sacrificed at (111 ± 1 days) . In this case, six age-matched FVB/NJ mice were used as the background wild-type control strain. The dissimilar backgrounds precluded direct comparisons between global SOD1G93A and MLC/SOD1G93A strains.
Dissociation of FDB fibers
The FDB muscles were dissected in cold (~4 °C) Rodent Ringer’s solution (in mM: 146 NaCl, 5 KCl, 2 CaCl2, 1 MgCl2, 10 HEPES, pH 7.4 with NaOH). Muscles were then digested in a mild collagenase solution (155 Cs-aspartate, 10 HEPES, 5 MgCl2, pH 7.4 with CsOH, supplemented with 1 mg/ml BSA, and 1 mg/ml collagenase type IA (Sigma-Aldrich, Saint Louis, MO) with agitation at 37 °C for ~1 h. Following digestion, the collagenase solution was replaced with dissociation solution (140 Cs-aspartate, 10 Cs2EGTA, 10 HEPES, 5 MgCl2, pH 7.4 with CsOH, supplemented with 1 mg/ml BSA) and muscles were triturated gently with a series of fire-polished glass Pasteur pipettes of descending bore. Dissociated FDB fibers destined for electrophysiological experiments were then plated onto ECL (Millipore, Billerica, MA)-coated 35-mm plastic culture dishes (Falcon, Tewksbury, MA). For Ca2+ imaging experiments, FDB fibers were plated onto laminin (Invitrogen, Eugene, OR)-coated 35-mm culture dishes with glass coverslip bottoms (MatTek, Ashland, MA). Experiments were performed with FDB fibers 1–6 h following dissociation.
Assessment of SR Ca2+ store content
Freshly dissociated FDB fibers were washed with Ca2+/Mg2+-free Ringer’s solution (in mM: 146 NaCl, 5 KCl, 10 HEPES, and 11 glucose, pH 7.4, with NaOH) twice and loaded with 5 μM Fluo 3-AM (Invitrogen) dissolved in Rodent Ringer’s solution for 20–30 min. Fibers were then washed three times in Rodent Ringer’s solution with gentle agitation. Fluo 3-AM-loaded fibers bathed in Rodent Ringer’s solution were then placed on the stage of an LSM META scanning laser confocal microscope (Carl Zeiss, Inc., Jena, Germany) and viewed under ×10 magnification. N-benzyl-p-toluenesulfonamide (BTS; 100 μM; Sigma-Aldrich) was always present in the bath solution (∼25 °C) to prevent contractions. The Fluo-3 dye was excited with the 488 nm line of an argon laser (30 mW maximum output, operated at 50 % or 6.3 A, attenuated to 1–2 %). The emitted fluorescence was directed to a photomultiplier equipped with a 505-nm long-pass filter. Confocal fluorescence intensity data were digitized at 8 bits, with the photomultiplier gain and offset adjusted such that maximum pixel intensities were no more than ∼70 % saturated and cell-free areas had close to zero intensity. Application of 4-chloro-m-cresol (4-CmC; Pfaltz and Bauer, Waterbury, CT) via a manually operated, gravity-driven global perfusion system was used to deplete the SR of Ca2+. Fluorescence amplitude data are expressed as ΔF/F, where F represents the baseline fluorescence before the application of 4-CmC, and ΔF represents the change in fluorescence during the application of 4-CmC.
Electrophysiology and whole-cell recordings of myoplasmic Ca2+ transients
The hindlimb muscles from wild-type and SOD1G93A mice (tibialis anterior, gastrocnemius, and FDB) were promptly removed following sacrifice, flash-frozen in liquid nitrogen, and stored at −80 °C for later use. At a later time, muscle tissue was homogenized by centrifugation with aluminum beads (NextAdvance, Averill Park, NY) in lysis buffer (50 mM Tris, 1 % SDS, Bio-Rad, Hercules, CA). Sample protein concentration was determined using the Bradley method (BCA assay kit, ThermoFisher, Pittsburgh, PA); lysates were divided into 30–50 μg aliquots. After addition of Laemmli buffer (Bio-Rad), proteins were separated by standard SDS-PAGE (10 % gels; Bio-Rad), transferred to nitrocellulose and non-specific immunoreactivity was blocked with 5 % non-fat dry milk (Kroger, Cincinnati, OH). Primary antibodies used for immunoblotting were mouse anti-CaV1.1 (also referred to as mAB 1A; 1:1000; ThermoScientific, Rockford, IL), mouse α-actin (1:1000; Sigma-Aldrich), and rabbit histone H3 (1:1000; Cell Signaling, Danvers, MA). Goat anti-mouse and anti-rabbit HRP-conjugated secondary antibodies were obtained from Southern Biotech (1:10000; Birmingham, AL). Blots were visualized with SuperSignal West Pico kit (ThermoFisher) viewed on a FluorChem HD2 scanner (Alpha Innotech, San Leandro, CA). Blots were stripped using Restore Western Blot Stripping Buffer (ThermoScientific). Cellular CaV1.1 protein levels measured using ImageJ densitometry (National Institutes of Health, Bethesda, MD) and were normalized to α-actin expression in the same sample (i.e., lane).
All data are presented as mean ± SEM. Statistical comparisons were made by two-tailed, unpaired t test with P < 0.05 considered significant. Figures were made using the software program SigmaPlot (version 11.0, SSPS Inc., San Jose, CA).
Results and discussion
EC coupling is impaired in symptomatic SOD1G93A FDB fibers
L-type currents and intramembrane charge movement are reduced in symptomatic SOD1G93A FDB fibers
Impaired CaV1.1 function in early-symptomatic SOD1G93A FDB fibers
EC coupling, L-type currents, and charge movement are reduced in FDB fibers expressing SOD1G93A autonomously in the skeletal muscle
Our experiments with early-symptomatic SOD1G93A mice raise the possibility that the reduction of CaV1.1 expression precedes overt neurological symptoms of ALS. Nonetheless, a degree of uncertainty exists concerning the contribution of denervation. On a more stringent level, the innervation profiles of the individual fibers used in our patch-clamp experiments were virtually impossible to assess accurately. To determine nerve-independent effects, we utilized a mouse model where transgenic expression of SOD1G93A is limited to skeletal muscle (MLC/SOD1G93A) . This mouse line enabled us to examine whether events occurring locally in skeletal muscle can cause impaired CaV1.1 function because substantial neuromuscular junction defects in such models do not develop until at least 8 months of age ([6, 7]; A.M., unpublished results).
In this study, we found that FDB fibers harvested from symptomatic global SOD1G93A mice had substantially reduced EC coupling relative to fibers of age-matched wild-type mice (Fig. 1). The impairment of EC coupling was not a consequence of a depleted SR Ca2+ store (Fig. 2), but appeared to be the result of reduced L-type Ca2+ channel membrane expression (though we cannot discount the presence of electrically silent channels). In particular, L-type Ca2+ current (Fig. 3a–c) and intramembrane charge movement (Fig. 3d–f) were both decreased by ~40 % in symptomatic SOD1G93A FDB fibers. The observed reductions in CaV1.1 function in symptomatic SOD1G93A fibers resembled the effect of application of ALS patient serum to normal rat extensor digitorum longus muscle [26, 27], but our results imply that these impairments develop more slowly over time.
Our data indicating that EC coupling and L-type current were compromised in symptomatic SOD1G93A muscle fibers contrast with those of another group who found that depolarization-dependent Ca2+ transients were slightly augmented in SOD1G93A mice (B6SJL-Tg(SOD1*G93A)1Gur/J) as a consequence of insufficient local mitochondrial Ca2+ buffering near the neuromuscular junction [40, 41]. Our findings also differ with those of another study which also examined Ca2+ handling in FDB fibers obtained from symptomatic SOD1G93A mice (B6.Cg-Tg(SOD1*G93A)1Gur/J). In this latter study , the authors observed virtually no difference in the amplitude of Ca2+ transients evoked by maximal field stimulation frequencies (100 Hz) using ratiometric Ca2+-sensitive dyes. It was concluded that the ability of CaV1.1 to engage EC coupling was normal in symptomatic SOD1G93A superficial gastrocnemius muscle because western blots showed no change in CaV1.1 expression . By contrast, our western blot analysis of whole-cell lysates obtained from symptomatic SOD1G93A tibialis anterior muscles indicated a clear reduction in total CaV1.1 expression (Fig. 4a). We did not observe significant differences for either gastrocnemius or FDB (Fig. 4b, c), though the band intensity difference in gastrocnemius was marginal (P = 0.074). The ambiguous nature of our biochemical results underscores the critical importance of our voltage-clamp approach to determine the relative number of functional CaV1.1 channels in the plasma membrane.
One reasonable explanation for the observed decreases in Ca2+ transient amplitude, peak L-type current density, and maximal charge movement is that SOD1G93A-expressing muscle undergoes fast-twitch (type IIb or IIx) to slow or intermediate (type I or type Ia, respectively) fiber-type switching [6, 30]. In an earlier study, slow-twitch rat soleus fibers were found to support voltage-dependent SR Ca2+ release only about half as well as fast-twitch fibers dissected from EDL of the same animals [43, 44]. Likewise, radioactive 1,4-dihydropyridine binding of the soleus was reduced ~50 % compared to that of EDL in the same study. In view of this difference in CaV1.1 expression between general fiber types, it is possible that we recorded from a population more heavily weighted in type I or type IIa fibers and that our data reflect fiber-type switching. Nevertheless, the dramatic alteration in the expression profile of CaV1.1 that accompanies fiber-type switching appears to be a downstream consequence of mutant SOD1 expression. Future experiments with more complex models that enable the distinction amongst fiber types in live cells are required to investigate this idea further.
Importantly, we also found that the impairment of CaV1.1 function in SOD1G93A mice began to develop prior the presentation of classical ALS symptoms. Though the reduction in depolarization-induced Ca2+ transients were not significantly reduced in early-symptomatic SOD1G93A muscle fibers (Fig. 5), decreased L-type current amplitude and intramembrane charge movement were clearly evident (Fig. 6). A number of explanations could account for this apparently discordant result. For example, the early-symptomatic SOD1G93A mice may have had a dissimilar RyR (type 1 or type 3) complement than the symptomatic mice or there have been a compensatory reversion to a juvenile RyR1 splice variant (ASI+) that supports greater SR Ca2+ release in response to depolarization [45, 46]. In any event, the results of our experiments with early-symptomatic SOD1G93A fibers suggested that the impairment of CaV1.1 function manifested in symptomatic SOD1G93A fibers arises, at least in part, from signals that originate in muscle (Figs. 5, 6, and 7). To test this idea, we assessed CaV1.1 function in fibers obtained from 4-month-old MLC/SOD1G93A mice in which expression of the mutant SOD1 protein is restricted to skeletal muscle. In these experiments, we observed ~35, ~20, and ~15 % reductions in Ca2+ transient amplitude, peak L-type current density, and maximal charge movement, respectively (Figs. 8 and 9). The reductions in L-type current amplitude and charge movement resembled the reductions in the parameters observed in early-symptomatic global SOD1G93A fibers. Interestingly, the impact of muscle-specific expression of SOD1G93A on EC coupling was greater than that observed in early-symptomatic global fibers. Previous ultrastructural analysis of the MLC/SOD1G93A strain suggests that concurrent uncoupling of triad junctions with transverse tubules may amplify the EC coupling impairment . Together, the ultrastructural and electrophysiological changes induced by muscle-specific expression of SOD1G93A can reasonably account for marked reductions in specific force observed in both fast- and slow-twitch hindlimb muscles of MLC/SOD1G93A mice . Thus, the results of our experiments with MLC/SOD1G93A fibers support the idea that muscle-specific events also make a significant contribution to ALS pathology, similar to what has been observed previously with myotonic dystrophy  and Huntington’s disease .
We did not observe a decrease in total membrane capacitance in SOD1G93A-expressing fibers, indicating that there was not substantial atrophy and/or loss of tubular surface area in the fibers that we examined electrophysiologically (Additional file 1: Figure S1). Based on the indisputable fact that atrophy is characteristic of both global and muscle-targeted SOD1G93A muscles, we think that our data are representative of a subpopulation of fibers healthy enough to survive the dissociation process and to meet the set visual criteria for voltage-clamp experiments (see Methods). If this is indeed the case, the reductions in voltage-dependent SR Ca2+ release, charge movement, and L-type current in the entire population of SOD1G93A fibers are likely more severe (and earlier onset) than we report.
Until recently, the involvement of CaV1.1 in the maintenance of muscle mass and composition was impossible to investigate because mice null for CaV1.1 (dysgenic) die immediately following parturition . To circumvent the perinatal lethality of CaV1.1 deletion, Piétri-Rouxel and colleagues delivered anti-CaV1.1 siRNA to the mouse hindlimb muscle with a serotype 1 Adeno-Associated Virus (AAV1) vector . Using this approach, they demonstrated that targeted, long-term suppression of L-type channel expression via induced exon-skipping produced prominent atrophy and extensive fibrosis. This atrophic effect of experimental downregulation of CaV1.1 expression suggested that pathological decreases in L-type channel activity, such as those we have observed in the two SOD1G93A models utilized in this study, may also lead to muscle degeneration and, possibly, contribute to the destabilization of neuromuscular junctions early in the progression of ALS. Our current work provides initial support for this idea, but further work is needed to reveal the precise mechanism. The first step will be to determine whether the downregulation of the channel and/or its auxiliary subunits occurs at the message level or post-translationally. For the former, quantification of CaV1.1 subunit message levels by qRT-PCR should be revealing. In regard to the latter, the chronic upregulation of expression the RGK family small GTP binding protein Rad (Ras-like Associated with Diabetes) — a potent constitutively active inhibitor of CaV1.1 [50, 51]—has been demonstrated in muscle of sporadic ALS patients of unknown aetiology and two established familial ALS mouse models (SOD1G93A and SOD1G86R) [52, 53]. The increases in muscle Rad expression were attributed to elevated cellular oxidative stress levels and coincided with the onset of spinal motoneuron death . Without being overspeculative, these observations raise the possibility that such chronic enhancement of muscle Rad expression may contribute to both atrophy and the dissolution of neuromuscular junctions in human ALS .
We report that myoplasmic Ca2+ transient amplitude, L-type current density, and intramembrane charge movement are progressively downregulated in flexor digitorum brevis (FDB) fibers obtained from SOD1G93A mice, the most extensively utilized mouse model of familial ALS. We also observed impairments of EC coupling, L-type current density, and charge movement in fibers of mice expressing SOD1G93A only in the skeletal muscle, signaling a muscle-specific contribution to SOD1G93A-induced downregulation of CaV1.1 expression and/or function. Since CaV1.1 is a positive regulator of muscle mass, our results suggest that altered CaV1.1 activity is a contributor to the muscle remodeling that occurs in ALS patients prior to the presentation of overt neurological symptoms.
4-CmC, 4-chloro-m-cresol; AAV1, serotype 1 adeno-associated virus; ALS, amyotrophic lateral sclerosis; EC, excitation-contraction; EDL, extensor digitorum longus; FDB, flexor digitorum brevis; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; IGF-1, insulin-like growth factor 1; RyR, ryanodine-sensitive intracellular Ca2+ release channel; SOD1, Cu/Zn superoxide dismutase 1; SR, sarcoplasmic reticulum; TA, tibialis anterior
We thank Drs. U. Meza and S. Papadopoulos for comments on an early draft of the manuscript. We are grateful to Drs. K.G. Beam and D. Oskar for the continued support. Confocal images were acquired in the University of Colorado-AMC Advanced Light Microscopy Core (funded in part by NIH/NCRR Colorado CTSI Grant Number UL1 RR025780).
This work was supported by grants from the Boettcher Foundation, the Judith and Jean Pape Adams Charitable Foundation (to R.A.B.), Fondazione Telethon and Fondazione Roma (to A.M.). D.B. received a stipend from 2T32 AG000279-11 (to R.S. Schwartz, University of Colorado School of Medicine, Department of Medicine-Geriatrics Division).
DB and RAB designed research, performed research, analyzed data, and wrote the paper. CFR performed experiments and analyzed data. AAV designed research and wrote the paper. GD and MM contributed unique materials. AM contributed unique materials, designed research, and wrote the paper. All authors read and approved the final manuscript.
The authors declare that they have no competing interests.
D.B. is a post-doctoral scientist in the Department of Medicine of the University of Colorado School of Medicine. C.F.R. is a production laboratory associate at GE Healthcare Dharmacon. G.D. is a post-doctoral scientist at the Institute Pasteur Cenci-Bolognetti, DAHFMO-Unit of Histology and Medical Embryology at La Sapienza University. M.M. is Ph.D. student at the Institute Pasteur Cenci-Bolognetti, DAHFMO-Unit of Histology and Medical Embryology at La Sapienza University. A.A.V. is an assistant professor in the Department of Biological Sciences at Wright State University. A.M. is an associate professor at the Institute Pasteur Cenci-Bolognetti, DAHFMO-Unit of Histology and Medical Embryology at La Sapienza University. R.A.B. is an assistant professor in the Department of Medicine of the University of Colorado School of Medicine.
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- Brown Jr RH. Amyotrophic lateral sclerosis: recent insights from genetics and transgenic mice. Cell. 1995;80:687–92.View ArticlePubMedGoogle Scholar
- de Belleroche J, Orrell R, King A. Familial amyotrophic lateral sclerosis/motor neurone disease (FALS): a review of current developments. J Med Genet. 1995;32:841–7.View ArticlePubMedPubMed CentralGoogle Scholar
- Rosen DR, Siddique T, Patterson D, Figelwicz DA, Sapp P, Hentati P, et al. Mutations in the Cu/Zn superoxide dismutase gene are associated with familial amyotrophic lateral sclerosis. Nature. 1993;362:59–62.View ArticlePubMedGoogle Scholar
- Gurney ME, Pu H, Chiu AY, Dal Canto MC, Polchow CY, Alexander DD, et al. Motor neuron degeneration in mice that express a human Cu, Zn superoxide dismutase mutation. Science. 1994;264:1772–5.View ArticlePubMedGoogle Scholar
- Dupuis L, Loeffler JP. Neuromuscular junction destruction during amyotrophic lateral sclerosis: insights from transgenic models. Curr Opin Pharmacol. 2009;9:341–6.View ArticlePubMedGoogle Scholar
- Dobrowolny G, Aucello M, Rizzuto E, Beccafico S, Mammucari C, Boncompagni S, et al. Skeletal muscle is a primary target of SOD1G93A-mediated toxicity. Cell Metab. 2008;8:425–36.View ArticlePubMedGoogle Scholar
- Wong M, Martin LJ. Skeletal muscle-restricted expression of human SOD1 causes motor neuron degeneration in transgenic mice. Hum Mol Genet. 2010;19:2284–302.View ArticlePubMedPubMed CentralGoogle Scholar
- Dadon-Nachum M, Melamed E, Offen D. The “dying-back” phenomenon of motor neurons in ALS. J Mol Neurosci. 2011;43:470–7.View ArticlePubMedGoogle Scholar
- Krakora D, Macrander C, Suzuki M. Neuromuscular junction protection for the potential treatment of amyotrophic lateral sclerosis. Neurol Res Int. 2012;2012:379657.PubMedPubMed CentralGoogle Scholar
- Musaró A. Understanding ALS: new therapeutic approaches. FEBS J. 2013;280:4315–22.View ArticlePubMedGoogle Scholar
- Pansarasa O, Rossi D, Berardinelli A, Cereda C. Amyotrophic lateral sclerosis and skeletal muscle: an update. Mol Neurobiol. 2013;49:984–90.View ArticlePubMedGoogle Scholar
- Dupuis L, de Aguilar Gonzalez JL, Echaniz-Laguna A, Eschbach J, Rene F, Oudart H, et al. Muscle mitochondrial uncoupling dismantles neuromuscular junction and triggers distal degeneration of motor neurons. PLoS One. 2009;4:e5390.View ArticlePubMedPubMed CentralGoogle Scholar
- Dobrowolny G, Giacinti C, Pelosi L, Nicoletti C, Winn N, Barberi L, et al. Muscle expression of a local IGF-1 isoform protects motor neurons in an ALS mouse model. J Cell Biol. 2005;168:193–9.View ArticlePubMedPubMed CentralGoogle Scholar
- Barberi L, Dobrowolny G, Pelosi L, Giacinti C, Musarò A. Muscle involvement and IGF-1 signaling in genetic disorders: new therapeutic approaches. Endocr Dev. 2009;14:29–37.View ArticlePubMedGoogle Scholar
- Krakora D, Mulcrone P, Meyer M, Lewis C, Bernau K, Gowing G, Zimprich C, Aebischer P, Svendsen CN, Suzuki M. Synergistic effects of GDNF and VEGF on lifespan and disease progression in a familial ALS rat model. Mol Ther. 2013;21:1602–10.View ArticlePubMedPubMed CentralGoogle Scholar
- Schneider MF, Chandler WK. Voltage dependent charge movement of skeletal muscle: a possible step in excitation-contraction coupling. Nature. 1973;242:244–6.View ArticlePubMedGoogle Scholar
- Ríos E, Brum G. Involvement of dihydropyridine receptors in excitation-contraction coupling in skeletal muscle. Nature. 1987;325:717–20.View ArticlePubMedGoogle Scholar
- Tanabe T, Beam KG, Powell JA, Numa S. Restoration of excitation-contraction coupling and slow Ca2+ current in dysgenic muscle by dihydropyridine receptor complementary DNA. Nature. 1988;336:134–9.View ArticlePubMedGoogle Scholar
- Bannister RA. Bridging the myoplasmic gap II: more recent advances in skeletal muscle EC coupling. J Exp Biol. 2015;219:175–82.View ArticleGoogle Scholar
- Cherednichenko G, Hurne AM, Fessenden JD, Lee EH, Allen PD, Beam KG, et al. Conformational activation of Ca2+ entry by depolarization of skeletal myotubes. Proc Natl Acad Sci U S A. 2004;101:15793–8.View ArticlePubMedPubMed CentralGoogle Scholar
- Bannister RA, Pessah IN, Beam KG. The skeletal L-type Ca2+ current is a major contributor to Excitation-Coupled Ca2+ Entry (ECCE). J Gen Physiol. 2009;133:79–91.View ArticlePubMedPubMed CentralGoogle Scholar
- Mosca B, Delbono O, Messi ML, Bergamelli I, Wang Z-M, Vukcevic M, et al. Enhanced dihydropyridine receptor calcium channel activity restores muscle strength in JP45/CASQ1 double knockout mice. Nat Commun. 2013;4:1541.View ArticlePubMedPubMed CentralGoogle Scholar
- Lee CS, Dagnino-Acosta A, Yarotskyy V, Hanna A, Lyfenko A, Knoblauch M, et al. Ca2+ permeation and/or binding to CaV1.1 fine-tunes skeletal muscle Ca2+ signaling to sustain muscle function. Skelet Muscle. 2015;5:4.View ArticlePubMedPubMed CentralGoogle Scholar
- Georgiou DK, Dagnino-Acosta A, Lee CS, Griffin DM, Wang H, Lagor WR, et al. Ca2+ binding/permeation via calcium channel, CaV1.1, regulates the intracellular distribution of the fatty acid transport protein, CD36, and fatty acid metabolism. J Biol Chem. 2015;290:23751–65.View ArticlePubMedGoogle Scholar
- Chen F, Liu Y, Sugiura Y, Allen PD, Gregg RG, Lin W. Neuromuscular synaptic patterning requires the function of skeletal muscle dihydropyridine receptors. Nat Neurosci. 2011;14:570–7.View ArticlePubMedPubMed CentralGoogle Scholar
- Delbono O, García J, Appel SH, Stefani E. IgG from amyotrophic lateral sclerosis affects tubular calcium channels of skeletal muscle. Am J Physiol. 1991;260:C1347–51.PubMedGoogle Scholar
- Delbono O, García J, Appel SH, Stefani E. Calcium current and charge movement of mammalian muscle: action of amyotrophic lateral sclerosis immunoglobulins. J Physiol. 1991;444:723–42.View ArticlePubMedPubMed CentralGoogle Scholar
- Magnelli V, Sawada T, Delbono O, Smith RG, Appel SH, Stefani E. The action of amyotrophic lateral sclerosis immunoglobulins on mammalian single skeletal muscle Ca2+ channels. J Physiol. 1993;461:103–18.View ArticlePubMedPubMed CentralGoogle Scholar
- Messi ML, Clark HM, Prevette DM, Oppenheim RW, Delbono O. The lack of effect of specific overexpression in the central nervous system or skeletal muscle on pathophysiology in the G93A SOD-1 mouse model of ALS. Exp Neurol. 2007;207:52–63.View ArticlePubMedPubMed CentralGoogle Scholar
- Hegedus J, Putman CT, Tyreman N, Gordon T. Preferential motor unit loss in the SOD1G93A transgenic mouse model of amyotrophic lateral sclerosis. J Physiol. 2008;586:3337–51.View ArticlePubMedPubMed CentralGoogle Scholar
- Hatzipetros T, Kidd JD, Moreno AJ, Thompson K, Gill A, Vieira FG. A quick phenotypic neurological scoring system for evaluating disease progression in the SOD1-G93A mouse model of ALS. J Vis Exp. 2015;104:e53257.Google Scholar
- Henriques A, Pitzer C, Schneider A. Characterization of a novel SOD-1(G93A) transgenic mouse line with very decelerated disease development. PLoS One. 2010;5:e15445.View ArticlePubMedPubMed CentralGoogle Scholar
- Delbono O, O’Rourke KS, Ettinger WH. Excitation-calcium release uncoupling in aged single human skeletal muscle fibers. J Membr Biol. 1995;148:211–22.View ArticlePubMedGoogle Scholar
- Jiménez-Moreno R, Wang ZM, Gerring RC, Delbono O. Sarcoplasmic reticulum Ca2+ release declines in muscle fibers from aging mice. Biophys J. 2008;94:3178–88.View ArticlePubMedPubMed CentralGoogle Scholar
- Taylor JR, Zheng Z, Wang ZM, Payne AM, Messi ML, Delbono O. Increased CaVβ1A expression with aging contributes to skeletal muscle weakness. Aging Cell. 2009;5:584–94.View ArticleGoogle Scholar
- Piétri-Rouxel F, Gentil C, Vassilopoulos S, Baas D, Mouisel E, Ferry A, et al. DHPR α1S subunit controls skeletal muscle mass and morphogenesis. EMBO J. 2010;29:643–54.View ArticlePubMedGoogle Scholar
- Beqollari D, Romberg CF, Filipova D, Meza U, Papadopoulos S, Bannister RA. Rem uncouples excitation-contraction coupling in adult mouse skeletal muscle fibers. J Gen Physiol. 2015;146:97–108.View ArticlePubMedPubMed CentralGoogle Scholar
- Vinsant S, Mansfield C, Jimenez-Moreno R, Del Gaizo Moore V, Yoshikawa M, Hampton TG, et al. Characterization of early pathogenesis in the SOD1 (G93A) mouse model of ALS: part I, background and methods. Brain Behav. 2013;4:335–50.View ArticleGoogle Scholar
- Vinsant S, Mansfield C, Jimenez-Moreno R, Del Gaizo Moore V, Yoshikawa M, Hampton TG, et al. Characterization of early pathogenesis in the SOD1 (G93A) mouse model of ALS: part II, results and discussion. Brain Behav. 2013;4:431–57.View ArticleGoogle Scholar
- Yi J, Ma C, Li Y, Weisleder N, Ríos E, Ma J, Zhou J. Mitochondrial calcium uptake regulates rapid calcium transients in skeletal muscle during excitation-contraction (E-C) coupling. J Biol Chem. 2011;286:32436–43.View ArticlePubMedPubMed CentralGoogle Scholar
- Zhou J, Yi J, Fu R, Liu E, Siddique T, Ríos E, et al. Hyperactive intracellular calcium signaling associated with localized mitochondrial defects in skeletal muscle of an animal model of amyotrophic lateral sclerosis. J Biol Chem. 2009;285:705–12.View ArticlePubMedPubMed CentralGoogle Scholar
- Chin ER, Chen D, Bobyk K, Mazala DA. Perturbations in intracellular Ca2+ handling in skeletal muscle in the G93A*SOD1 mouse model of amyotrophic lateral sclerosis. Am J Physiol-Cell Physiol. 2014;307:C1031–8.View ArticlePubMedPubMed CentralGoogle Scholar
- Delbono O, Meissner G. Sarcoplasmic reticulum Ca2+ release in rat slow- and fast-twitch muscles. J Membr Biol. 1996;151:123–30.View ArticlePubMedGoogle Scholar
- Payne AM, Delbono O. Neurogenesis of excitation-contraction uncoupling in aging skeletal muscle. Exerc Sport Sci Rev. 2004;32:36–40.View ArticlePubMedGoogle Scholar
- Futatsugi A, Kuwajima G, Mikoshiba K. Tissue-specific and developmentally regulated alternative splicing in mouse skeletal muscle ryanodine receptor mRNA. Biochem J. 1995;305:373–8.View ArticlePubMedPubMed CentralGoogle Scholar
- Kimura T, Lueck JD, Harvey PJ, Pace SM, Ikemoto N, Casarotto MG, et al. Alternative splicing of RyR1 alters the efficacy of skeletal EC coupling. Cell Calcium. 2009;45:264–74.View ArticlePubMedPubMed CentralGoogle Scholar
- Tang ZZ, Yarotskyy V, Wei L, Sobczak K, Nakamori M, Eichinger K, Moxley RT, Dirksen RT, Thornton CA. Muscle weakness in myotonic dystrophy associated with misregulated splicing and altered gating of CaV1.1 calcium channel. Hum Mol Genet. 2012;21:1312–24.View ArticlePubMedGoogle Scholar
- Waters CW, Varuzhanyan G, Talmadge RJ, Voss AA. Huntington disease skeletal muscle is hyperexcitable owing to chloride and potassium channel dysfunction. Proc Natl Acad Sci U S A. 2013;110:9160–5.View ArticlePubMedPubMed CentralGoogle Scholar
- Beam KG, Knudson CM, Powell JA. A lethal mutation in mice eliminates the slow calcium current in skeletal muscle cells. Nature. 1986;320:168–70.View ArticlePubMedGoogle Scholar
- Romberg CF, Beqollari D, Meza U, Bannister RA. RGK proteins inhibit slow depolarization-dependent Ca2+ entry in developing myotubes. Channels. 2014;8:243–8.View ArticlePubMedPubMed CentralGoogle Scholar
- Beqollari D, Romberg CF, Meza U, Papadopoulos S, Bannister RA. Differential effects of RGK proteins on L-type Ca2+ channel function in mouse skeletal muscle. Biophys J. 2014;106:1950–7.View ArticlePubMedPubMed CentralGoogle Scholar
- de Aguilar Gonzalez JL, Niederhauser-Wiederkehr C, Halter B, De Tapia M, Di Scala F, Demougin P, et al. Gene profiling of skeletal muscle in an amyotrophic lateral sclerosis mouse model. Physiol Genomics. 2008;32:207–18.View ArticleGoogle Scholar
- Halter B, de Aguilar Gonzalez JL, Rene F, Petri S, Fricker B, Echaniz-Laguna A, et al. Oxidative stress in skeletal muscle stimulates early expression of Rad in a mouse model of amyotrophic lateral sclerosis. Free Radic Biol Med. 2010;48:915–23.View ArticlePubMedGoogle Scholar