Molecular characterization of the intact mouse muscle spindle using a multi-omics approach

Bornstein, Bavat; Heinemann-Yerushalmi, Lia; Krief, Sharon; Adler, Ruth; Dassa, Bareket; Leshkowitz, Dena; Kim, Min‐Chul; Bewick, Guy S.; Banks, Robert W.; Zelzer, Elazar

doi:10.7554/elife.81843

Cited by 11 publications

(19 citation statements)

References 55 publications

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“…For whole-mount immunofluorescence, muscles were subjected to an optical tissue clearing protocol as described previously (Bornstein et al, 2023) Next, samples were washed in PBS, transferred to 5 mL of 10% SDS (pH 8.0) with 0.01% sodium azide, and shaken gently at 37 • C for 3 days to remove lipid.…”

Section: Immunofluorescencementioning

confidence: 99%

The mechanosensitive ion channel ASIC2 mediates both proprioceptive sensing and spinal alignment

Bornstein

Watkins²,

Passini

et al. 2023

Experimental Physiology

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New Findings What is the central question of this study? Proprioception is initiated by mechanosensitive neurons. However, the identities of the molecular players that mediate proprioceptive sensing are largely unknown. Here, we aimed to identify potential mechanosensitive ion channels that mediate proprioceptive signalling. What is the main finding and its importance? We identify the mechanosensitive ion channel ASIC2 as a key component in proprioceptive sensing and a regulator of spine alignment. AbstractBy translating mechanical forces into molecular signals, proprioceptive neurons provide the CNS with information on muscle length and tension, which is necessary to control posture and movement. However, the identities of the molecular players that mediate proprioceptive sensing are largely unknown. Here, we confirm the expression of the mechanosensitive ion channel ASIC2 in proprioceptive sensory neurons. By combining in vivo proprioception‐related functional tests with ex vivo electrophysiological analyses of muscle spindles, we showed that mice lacking Asic2 display impairments in muscle spindle responses to stretch and motor coordination tasks. Finally, analysis of skeletons of Asic2 loss‐of‐function mice revealed a specific effect on spinal alignment. Overall, we identify ASIC2 as a key component in proprioceptive sensing and a regulator of spine alignment.

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Section: Immunofluorescencementioning

confidence: 99%

The mechanosensitive ion channel ASIC2 mediates both proprioceptive sensing and spinal alignment

Bornstein

Watkins²,

Passini

et al. 2023

Experimental Physiology

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“…To investigate the extracellular matrix and the integrity of the connective tissue capsule of muscle spindles from wildtype and mutant animals, we compared the distribution of the matrix protein versican in Gaa −/− mice and in age-matched 129/SvJ control mice. In skeletal muscle, anti-versican antibodies selectively label the muscle spindle extracellular matrix 26 . Versican immunoreactivity was restricted to the inner and outer capsule in muscle spindles from wildtype mice (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Antibodies against the lysosomal membrane glycoprotein LAMP1 (L1418, Sigma-Aldrich, Darmstadt, Germany; 1:500), which plays an important role in lysosome biogenesis and autophagy, were used to investigate lysosomal buildup. Versican was detected using a rabbit anti-versican antibody (Ab19345; Abcam, Cambridge, UK; 1:500 26 ). The distribution of the voltage-gated sodium channel was analyzed using a polyclonal rabbit antibody (SCN4A; #ASC-020; Alomone labs, Jerusalem, Israel; 1:500 25 ).…”

Section: Methodsmentioning

confidence: 99%

Degeneration of muscle spindles in a murine model of Pompe disease

Watkins¹,

Schultheiß²,

Rafuna³

et al. 2023

Sci Rep

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Pompe disease is a debilitating medical condition caused by a functional deficiency of lysosomal acid alpha-glucosidase (GAA). In addition to muscle weakness, people living with Pompe disease experience motor coordination deficits including an instable gait and posture. We reasoned that an impaired muscle spindle function might contribute to these deficiencies and therefore analyzed proprioception as well as muscle spindle structure and function in 4- and 8-month-old Gaa−/− mice. Gait analyses showed a reduced inter-limb and inter-paw coordination in Gaa−/− mice. Electrophysiological analyses of single-unit muscle spindle proprioceptive afferents revealed an impaired sensitivity of the dynamic and static component of the stretch response. Finally, a progressive degeneration of the sensory neuron and of the intrafusal fibers was detectable in Gaa−/− mice. We observed an increased abundance and size of lysosomes, a fragmentation of the inner and outer connective tissue capsule and a buildup of autophagic vacuoles in muscle spindles from 8-month-old Gaa−/− mice, indicating lysosomal defects and an impaired autophagocytosis. These results demonstrate a structural and functional degeneration of muscle spindles and an altered motor coordination in Gaa−/− mice. Similar changes could contribute to the impaired motor coordination in patients living with Pompe disease.

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“…The proteome of intrafusal muscle spindles, which run anatomically in parallel to bulk extrafusal myofibers, are part of the proprioceptive system of the body that is intrinsically involved in controlling skeletal muscle length, coordinated movements and posture, was recently analyzed [295]. As compared to skeletal muscle fibers, the proprioceptive muscle system was shown to express higher levels of myosins-3/6/7/7b, collagen isoform COL-IV and sensory neuron proteins PIEZO2 and SLC17A7.…”

Section: The Proteome Of Specialized Cells and Structures Within Skel...mentioning

confidence: 99%

“…The proteomic information summarizes the methodological approaches taken to establish the core protein components of skeletal muscles. The proteomic studies and reviews listed include (i) the initiation of the skeletal muscle proteome project [21,22], (ii) the systematic cataloguing of human skeletal muscles [20,23,26,27,37,53], (iii) proteomic surveys of fast versus slow human muscles [29,292], (iv) the analysis of human single myofibers [35,36,313], (v) the systematic cataloguing of animal skeletal muscles [25,28,288,290,[328][329][330], (vi) the differential proteomic surveys of fast versus slow mouse muscles [30][31][32], (vii) profiling of animal single myofibers [25,33,277,311,312], (viii) the mass spectrometric analysis of muscle spindles [295], (ix) the profiling of the neuromuscular junction [296,297], (x) proteomic profiling of subcellular fractions (sarcolemma, sarcoplasmic reticulum, mitochondria, sarcosol, giant muscle protein assemblies, contractile apparatus) [92,140,175,[304][305][306][308][309][310], (xi) the systematic cataloguing of the muscle secretome…”

Section: Progress Of Cataloguing the Skeletal Muscle Proteomementioning

confidence: 99%

Mass Spectrometry-Based Proteomic Technology and Its Application to Study Skeletal Muscle Cell Biology

Dowling,

Swandulla,

Ohlendieck

2023

Cells

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Voluntary striated muscles are characterized by a highly complex and dynamic proteome that efficiently adapts to changed physiological demands or alters considerably during pathophysiological dysfunction. The skeletal muscle proteome has been extensively studied in relation to myogenesis, fiber type specification, muscle transitions, the effects of physical exercise, disuse atrophy, neuromuscular disorders, muscle co-morbidities and sarcopenia of old age. Since muscle tissue accounts for approximately 40% of body mass in humans, alterations in the skeletal muscle proteome have considerable influence on whole-body physiology. This review outlines the main bioanalytical avenues taken in the proteomic characterization of skeletal muscle tissues, including top-down proteomics focusing on the characterization of intact proteoforms and their post-translational modifications, bottom-up proteomics, which is a peptide-centric method concerned with the large-scale detection of proteins in complex mixtures, and subproteomics that examines the protein composition of distinct subcellular fractions. Mass spectrometric studies over the last two decades have decisively improved our general cell biological understanding of protein diversity and the heterogeneous composition of individual myofibers in skeletal muscles. This detailed proteomic knowledge can now be integrated with findings from other omics-type methodologies to establish a systems biological view of skeletal muscle function.

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Molecular characterization of the intact mouse muscle spindle using a multi-omics approach

Cited by 11 publications

References 55 publications

The mechanosensitive ion channel ASIC2 mediates both proprioceptive sensing and spinal alignment

The mechanosensitive ion channel ASIC2 mediates both proprioceptive sensing and spinal alignment

Degeneration of muscle spindles in a murine model of Pompe disease

Mass Spectrometry-Based Proteomic Technology and Its Application to Study Skeletal Muscle Cell Biology

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