Spinal muscular atrophy (SMA) is a neuromuscular genetic disease caused by reduced survival motor neuron (SMN) protein. SMN is ubiquitous and deficient levels cause spinal cord motoneurons (MNs) degeneration and muscle atrophy. Nevertheless, the mechanism by which SMN reduction in muscle contributes to SMA disease is not fully understood. Therefore, studies evaluating atrophy mechanisms in SMA muscles will contribute to strengthening current knowledge of the pathology. Here we propose to evaluate autophagy in SMA muscle, a pathway altered in myotube atrophy. We analized autophagy proteins and mTOR in muscle biopsies, fibroblasts, and lymphoblast cell lines from SMA patients and in gastrocnemius muscles from a severe SMA mouse model. Human MNs differentiated from SMA and unaffected control iPSCs were also included in the analysis of the autophagy. Muscle biopsies, fibroblasts, and lymphoblast cell lines from SMA patients showed reduction of the autophagy marker LC3-II. In SMA mouse gastrocnemius, we observed lower levels of LC3-II, Beclin 1, and p62/SQSTM1 proteins at pre-symptomatic stage. mTOR phosphorylation at Ser2448 was decreased in SMA muscle cells. However, in mouse and human cultured SMA MNs mTOR phosphorylation and LC3-II levels were increased. These results suggest a differential regulation in SMA of the autophagy process in muscle cells and MNs. Opposite changes in autophagy proteins and mTOR phosphorylation between muscle cells and neurons were observed. These differences may reflect a specific response to SMN reduction, which could imply diverse tissue-dependent reactions to therapies that should be taken into account when treating SMA patients.
Abstract--Experimental alteration of volcanic tuff from Almeria, southeastern Spain, was performed in solutions with different Na/K ratios (0.01, 1, 10, and 100), different total salt concentrations (0.01, 0.1, 0.2, 0.33, and 1 M), and in deionized water, at 60, 80, 120, and 160~ for periods of 60, 90, 180, and 360 d. Two particle size fractions of volcanic tuff were used: 10-200 and 20-60 p,m. Alteration products were examined by X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), laser-particle size analysis, scanning electron microscopy equipped with an energy dispersive X-ray spectrometer (SEM-EDS), image computer analysis, and transmission electron microscopy with microanalysis (TEM-AEM). XRD detected neoformed phases only in the products from experiments of 180-360 d at high temperatures (120-160~and with Na/K ratios above unity and in deionized water. The synthesized phase is a random mixed-layer illite-smectite (I-S) with 75% smectite. The quantity of newly formed I-S, determined by FTIR, ranged between 3-30%. There was no apparent change in grain size and shape of the grains after the experiments as compared to before.SEM-EDS and TEM-AEM revealed the following alteration sequence: 1) intense etching on glassgrain surfaces; 2) formation of hemispherical morphologies on grain surfaces; 3) precipitation of very thin, individual flakes of illite-smectite on glass-grain surfaces; 4) development of I-S at the edges of glass grains; and 5) development of I-S honeycomb structures either covering large areas of the glass grains or resulting from the complete alteration of glass grains. A direct transformation of glass to I-S seems to be the major reaction mechanism, although there also is evidence of glass dissolution and subsequent I-S precipitation.
Spinal muscular atrophy (SMA), a leading genetic cause of infant death, is caused by the loss of survival motor neuron 1 ( SMN1 ) gene. SMA is characterized by the degeneration and loss of spinal cord motoneurons (MNs), muscular atrophy, and weakness. SMN2 is the centromeric duplication of the SMN gene, whose numbers of copies determine the intracellular levels of SMN protein and define the disease onset and severity. It has been demonstrated that elevating SMN levels can be an important strategy in treating SMA and can be achieved by several mechanisms, including promotion of protein stability. SMN protein is a direct target of the calcium-dependent protease calpain and induces its proteolytic cleavage in muscle cells. In this study, we examined the involvement of calpain in SMN regulation on MNs. In vitro experiments showed that calpain activation induces SMN cleavage in CD1 and SMA mouse spinal cord MNs. Additionally, calpain 1 knockdown or inhibition increased SMN level and prevent neurite degeneration in these cells. We examined the effects of calpain inhibition on the phenotype of two severe SMA mouse models. Treatment with the calpain inhibitor, calpeptin, significantly improved the lifespan and motor function of these mice. Our observations show that calpain regulates SMN level in MNs and calpeptin administration improves SMA phenotype demonstrating the potential utility of calpain inhibitors in SMA therapy.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.