Linking cytoarchitecture to metabolism: sarcolemma-associated plectin affects glucose uptake by destabilizing microtubule networks in mdx myofibers

BACKGROUND: Duchenne muscular dystrophy (DMD) is one of the most frequent forms of muscular disorders. It is caused by the absence of dystrophin, a core component of the sarcolemma-associated junctional complex that links the cytoskeleton to the extracellular matrix. We showed previously that plectin 1f (P1f), one of the major muscle-expressed isoforms of the cytoskeletal linker protein plectin, accumulates at the sarcolemma of DMD patients as well as of mdx mice, a widely studied animal model for DMD.Based on plectin's dual role as structural protein and scaffolding platform for signaling molecules, we speculated that the dystrophic phenotype observed after loss of dystrophin was caused, at least to some extent, by excess plectin. Thus, we hypothesized that elimination of plectin expression in mdx skeletal muscle, while probably resulting in an overall more severe phenotype, may lead to a partial phenotype rescue. In particular, we wanted to assess whether excess sarcolemmal plectin contributes to the dysregulation of sugar metabolism in mdx myofibers. METHODS: We generated plectin/dystrophin double deficient (dKO) mice by breeding mdx with conditional striated muscle-restricted plectin knockout (cKO) mice. The phenotype of these mice was comparatively analyzed with that of mdx, cKO, and wild-type mice, focusing on structural integrity and dysregulation of glucose metabolism. RESULTS: We show that the accumulation of plectin at the sarcolemma of mdx muscle fibers hardly compensated for their loss of structural integrity. Instead, it led to an additional metabolic deficit by impairing glucose uptake. While dKO mice suffered from an overall more severe form of muscular dystrophy compared to mdx or plectin-deficient mice, sarcolemmal integrity as well as glucose uptake of their myofibers were restored to normal levels upon ablation of plectin. Furthermore, microtubule (MT) networks in intact dKO myofibers, including subsarcolemmal areas, were found to be more robust than those in mdx mice. Finally, myotubes differentiated from P1f-overexpressing myoblasts showed an impairment of glucose transporter 4 translocation and a destabilization of MT networks. CONCLUSIONS: Based on these results we propose that sarcolemma-associated plectin acts as an antagonist of MT network formation in myofibers, thereby hindering vesicle-mediated (MT-dependent) transport of glucose transporter 4. This novel role of plectin throws a bridge between extra-sarcomeric cytoarchitecture and metabolism of muscle fibers. Our study thus provides new insights into pathomechanisms of plectinopathies and muscular dystrophies in general

The certificate course "Data Librarian" and its first implementation

Im digitalen Zeitalter bieten Bibliotheken als Serviceeinrichtungen, neben den gedruckten Beständen vor Ort, in immer größerem Maße den Zugang zu elektronischen Ressourcen und digitalen Inhalten an. Durch den digitalen Wandel und die rasanten Entwicklungen im Informationsbereich ergeben sich neue Herausforderungen und Tätigkeitsfelder. Um den Anforderungen des Berufsfeldes gerecht zu werden, bedarf es einer permanenten Fort- und Weiterbildung. Infolgedessen wurde der Zertifikatskurs „Data Librarian“ ins Leben gerufen, der Kenntnisse, die für Entwicklung und Umsetzung von Services im Bereich Forschungsdatenmanagement von Nutzen sind, vermitteln soll. Schwerpunkte dieses Zertifikatskurses sind sind Themenbereiche Wissenschaftskommunikation und Forschungsunterstützung, Policies im Umgang mit Forschungsdaten, Datenmanagementpläne, Metadaten im Bereich Repositorien, Datenanalyse, Datenaggregation und Verlinkung, Datenstandards, Datenmodellierung, Langzeitarchivierung und Datensicherung. Im vorliegenden Beitrag berichten OrganisatorInnen, Vortragende und TeilnehmerInnen über Erfahrungen, die sie im Zuge der erstmaligen Durchführung des Kurses gemacht haben.In the digital age, libraries as service institutions are increasingly providing access to electronic resources and digital content, in addition to the printed collections on site. Digital change and rapid developments in the information sector are creating new challenges and new fields of activity. In order to meet the requirements of the professional field, permanent further education and training is required. As a result, the certificate course „Data Librarian“ was created to impart knowledge that is useful for the development and implementation of services in the field of research data management. The main focus of this certificate course is on scholarly communication and research support, policies for handling research data, data management plans, metadata in the field of repositories, data analysis, data aggregation and linking, data standards, data modelling, long-term preservation and data protection. In this article, organisers, lecturers and participants report on their experiences during the first implementation of the course

Plectin 1f scaffolding at the sarcolemma of dystrophic (mdx) muscle fibers through multiple interactions with β-dystroglycan

In skeletal muscle, the cytolinker plectin is prominently expressed at Z-disks and the sarcolemma. Alternative splicing of plectin transcripts gives rise to more than eight protein isoforms differing only in small N-terminal sequences (5–180 residues), four of which (plectins 1, 1b, 1d, and 1f) are found at substantial levels in muscle tissue. Using plectin isoform–specific antibodies and isoform expression constructs, we show the differential regulation of plectin isoforms during myotube differentiation and their localization to different compartments of muscle fibers, identifying plectins 1 and 1f as sarcolemma-associated isoforms, whereas plectin 1d localizes exclusively to Z-disks. Coimmunoprecipitation and in vitro binding assays using recombinant protein fragments revealed the direct binding of plectin to dystrophin (utrophin) and β-dystroglycan, the key components of the dystrophin–glycoprotein complex. We propose a model in which plectin acts as a universal mediator of desmin intermediate filament anchorage at the sarcolemma and Z-disks. It also explains the plectin phenotype observed in dystrophic skeletal muscle of mdx mice and Duchenne muscular dystrophy patients

Academic Library Education at the Universities of Vienna, Graz and Innsbruck in Cooperation with the Austrian National Library

Dieser Beitrag bietet einen Überblick über die Aus- und Fortbildung im wissenschaftlichen Bibliothekswesen Österreichs in den Jahren 2019 und 2020.This article provides an overview of the education and training in the academic library sector in Austria in 2019 and 2020

Academic library education and training in Austria 2020–2021

Dieser Beitrag bietet einen Überblick über die Aus- und Fortbildung im wissenschaftlichen Bibliothekswesen Österreichs in den Jahren 2020 und 2021.This article provides an overview of the education and training in the academic library sector in Austria in 2020 and 2021

Relationship between myosin heavy chain fibre type and restoration of dystrophin expression and key components of the dystrophin-associated glycoprotein complex by Tricyclo-DNA mediated exon skipping

Exon skipping mediated by tricyclo-DNA (tc-DNA) antisense oligonucleotides has been shown to induce significant levels of dystrophin restoration in mdx, a mouse model of Duchenne Muscular Dystrophy. This translates into significant improvement in key disease indicators in muscle, cardio-respiratory function, heart and the central nervous system. Here we examine the relationship between muscle fibre type, based on Myosin Heavy chain profile, and the ability of tc-DNA to restore not only dystrophin but also other members of the dystrophin-associated glycoprotein complex (DAPC). We first profiled this relationship in untreated mdx muscle and found that all fibre types support reversion events to a dystrophin positive state, in an unbiased manner. Importantly, we show that only a small fraction of revertant fibres expressed other members of the DAPC. Immunoblot analysis of protein levels, however, revealed robust expression of these components, which failed to correctly localise to the sarcolemma. We then show that tc-DNA treatment leads to nearly all fibres expressing not only dystrophin but also other key components of the DAPC. Of significance, our work shows that MHC fibre type does not bias the expression of any of these important proteins. This work also highlights that the improved muscle physiology following tc-DNA treatment reported previously results from the complete restoration of the dystrophin complex in all MHCII fibres with equal efficiencies

From a literature review to a conceptual framework for health sector websites’ assessment

Health sector institutions’ websites need to act as effective web resources of information and interactive communication mediums to address the versatile demands of their multiple stakeholders. Academic and practitioner interest in health sector website assessment has considerably risen in recent years. This can be seen by the number of papers published in journals. The purpose of this paper is twofold to further establish the field. First, it offers a literature re-view on hospitals’ websites assessment. Second, it offers a conceptual framework to address the website assessment issue in health sector. The proposed assessment framework focuses on four main criteria: content, technology, services, and participation being evaluated by the use of several indicators. Academics, hospital practitioners, public officials and users will find the review and the framework useful, as they outline major lines of research in the field and a method to assess health institution websites.This paper is a result of the project “SmartEGOV: Harnessing EGOV for Smart Governance (Foundations, methods, Tools) / NORTE-01-0145-FEDER-000037”, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (EFDR).info:eu-repo/semantics/publishedVersio

Keeping the Vimentin Network under Control: Cell–Matrix Adhesion–associated Plectin 1f Affects Cell Shape and Polarity of Fibroblasts

Mature focal adhesions and fibrillar adhesions act as anchorage sites for vimentin filaments, with plectin isoform 1f being the crucial linker protein. Plectin serves as a nucleation and assembly center for the de novo formation of vimentin networks. Anchored vimentin creates a resilient cage-like core structure that affects cell shape

Preventing phosphorylation of dystroglycan ameliorates the dystrophic phenotype in mdx mouse

Loss of dystrophin protein due to mutations in the DMD gene causes Duchenne muscular dystrophy. Dystrophin loss also leads to the loss of the dystrophin glycoprotein complex (DGC) from the sarcolemma which contributes to the dystrophic phenotype. Tyrosine phosphorylation of dystroglycan has been identified as a possible signal to promote the proteasomal degradation of the DGC. In order to test the role of tyrosine phosphorylation of dystroglycan in the aetiology of DMD, we generated a knock-in mouse with a phenylalanine substitution at a key tyrosine phosphorylation site in dystroglycan, Y890. Dystroglycan knock-in mice (Dag1Y890F/Y890F) had no overt phenotype. In order to examine the consequence of blocking dystroglycan phosphorylation on the aetiology of dystrophin-deficient muscular dystrophy, the Y890F mice were crossed with mdx mice an established model of muscular dystrophy. Dag1Y890F/Y890F/mdx mice showed a significant improvement in several parameters of muscle pathophysiology associated with muscular dystrophy, including a reduction in centrally nucleated fibres, less Evans blue dye infiltration and lower serum creatine kinase levels. With the exception of dystrophin, other DGC components were restored to the sarcolemma including α-sarcoglycan, α-/β-dystroglycan and sarcospan. Furthermore, Dag1Y890F/Y890F/mdx showed a significant resistance to muscle damage and force loss following repeated eccentric contractions when compared with mdx mice. While the Y890F substitution may prevent dystroglycan from proteasomal degradation, an increase in sarcolemmal plectin appeared to confer protection on Dag1Y890F/Y890F/mdx mouse muscle. This new model confirms dystroglycan phosphorylation as an important pathway in the aetiology of DMD and provides novel targets for therapeutic intervention