Proteome-wide Changes in the mdx-4cv Spleen due to Pathophysiological Cross Talk with Dystrophin-Deficient Skeletal Muscle

Dowling, Paul; Gargan, Stephen; Zweyer, Margit; Henry, Michael; Swandulla, Dieter; Ohlendieck, Kay

doi:10.1016/j.isci.2020.101500

Cited by 23 publications

(44 citation statements)

References 80 publications

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“…Current techniques are highly efficient and allow for the rapid analysis of many sample types. This protocol represents a typical bottom-up proteomic workflow and was used in a recent publication ( Dowling et al., 2020 ) to identify proteome-wide changes in the spleen of the dystrophic mdx-4cv mouse model of Duchenne muscular dystrophy. In Figure 1 , the dissection of mouse spleen is shown.…”

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confidence: 99%

“…Progressive dysfunction has also been observed in a variety of other organs, such as the heart, brain, kidney, and liver ( Murphy et al., 2018 ). The protocol described here using bottom-up proteomics to study detailed changes in a specific organ, i.e., the mass spectrometric analysis of the spleen from the mdx-4cv mouse ( Dowling et al., 2020 ), has elucidated the importance of organ crosstalk in X-linked muscular dystrophy. This pathophysiological complexity of body-wide alterations makes the mdx-4cv mutant an appropriate mouse model to investigate proteome-wide changes in dystrophinopathy.…”

Section: Before You Beginmentioning

confidence: 99%

“…In contrast, discovery proteomics focuses on large-scale protein screening and the analysis of protein dynamics in complex cellular systems. In this protocol, discovery proteomics was chosen to analyze differences in the protein expression profile of the mdx-4cv spleen ( Dowling et al., 2020 ). For comparative analyses, proteomics, in combination with bioinformatics, can be used to swiftly determine spatial and temporal alterations in protein expression patterns.…”

Section: Before You Beginmentioning

confidence: 99%

“…We then describe the steps for mass spectrometric analysis and bioinformatic evaluation. For complete details on the use and execution of this protocol, please refer to Dowling et al. (2020 ).…”

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“…For complete details on the use and execution of this protocol, please refer to Dowling et al. (2020 ).…”

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Protocol for the Bottom-Up Proteomic Analysis of Mouse Spleen

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Summary This protocol describes the comparative proteomic profiling of the spleen of wild type versus mdx-4cv mouse, a model of dystrophinopathy. We detail sample preparation for bottom-up proteomic mass spectrometry experiments, including homogenization of tissue, protein concentration measurements, protein digestion, and removal of interfering chemicals. We then describe the steps for mass spectrometric analysis and bioinformatic evaluation. For complete details on the use and execution of this protocol, please refer to Dowling et al. (2020 ).

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confidence: 99%

Section: Before You Beginmentioning

confidence: 99%

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confidence: 99%

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“…For complete details on the use and execution of this protocol, please refer to Dowling et al. (2020 ).…”

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Protocol for the Bottom-Up Proteomic Analysis of Mouse Spleen

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Proteomic profiling of impaired excitation–contraction coupling and abnormal calcium handling in muscular dystrophy

et al. 2022

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The X-linked inherited neuromuscular disorder Duchenne muscular dystrophy is characterised by primary abnormalities in the membrane cytoskeletal component dystrophin. The almost complete absence of the Dp427-M isoform of dystrophin in skeletal muscles renders contractile fibres more susceptible to progressive degeneration and a leaky sarcolemma membrane. This in turn results in abnormal calcium homeostasis, enhanced proteolysis and impaired excitation-contraction coupling. Biochemical and mass spectrometry-based proteomic studies of both patient biopsy specimens and genetic animal models of dystrophinopathy have demonstrated significant changes in the concentration and/or physiological function of essential calciumregulatory proteins in dystrophin-lacking voluntary muscles. Abnormalities include dystrophinopathy-associated changes in voltage sensing receptors, calcium release channels, calcium pumps and calcium binding proteins. This review article provides an overview of the importance of the sarcolemmal dystrophin-glycoprotein complex and the wider dystrophin complexome in skeletal muscle and its linkage to depolarisationinduced calcium-release mechanisms and the excitation-contraction-relaxation cycle.Besides chronic inflammation, fat substitution and reactive myofibrosis, a major pathobiochemical hallmark of X-linked muscular dystrophy is represented by the chronic influx of calcium ions through the damaged plasmalemma in conjunction with abnormal intracellular calcium fluxes and buffering. Impaired calcium handling proteins should therefore be included in an improved biomarker signature of Duchenne muscular dystrophy.

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Complexity of skeletal muscle degeneration: multi-systems pathophysiology and organ crosstalk in dystrophinopathy

Ohlendieck

Swandulla

2021

Pflugers Arch - Eur J Physiol

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Duchenne muscular dystrophy is a highly progressive muscle wasting disorder due to primary abnormalities in one of the largest genes in the human genome, the DMD gene, which encodes various tissue-specific isoforms of the protein dystrophin. Although dystrophinopathies are classified as primary neuromuscular disorders, the body-wide abnormalities that are associated with this disorder and the occurrence of organ crosstalk suggest that a multi-systems pathophysiological view should be taken for a better overall understanding of the complex aetiology of X-linked muscular dystrophy. This article reviews the molecular and cellular effects of deficiency in dystrophin isoforms in relation to voluntary striated muscles, the cardio-respiratory system, the kidney, the liver, the gastrointestinal tract, the nervous system and the immune system. Based on the establishment of comprehensive biomarker signatures of X-linked muscular dystrophy using large-scale screening of both patient specimens and genetic animal models, this article also discusses the potential usefulness of novel disease markers for more inclusive approaches to differential diagnosis, prognosis and therapy monitoring that also take into account multi-systems aspects of dystrophinopathy. Current therapeutic approaches to combat muscular dystrophy are summarised.

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Proteome-wide Changes in the mdx-4cv Spleen due to Pathophysiological Cross Talk with Dystrophin-Deficient Skeletal Muscle

Cited by 23 publications

References 80 publications

Protocol for the Bottom-Up Proteomic Analysis of Mouse Spleen

Protocol for the Bottom-Up Proteomic Analysis of Mouse Spleen

Proteomic profiling of impaired excitation–contraction coupling and abnormal calcium handling in muscular dystrophy

Complexity of skeletal muscle degeneration: multi-systems pathophysiology and organ crosstalk in dystrophinopathy

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