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Novel tissue-specific markers revealed by in-depth proteome analysis of human myotendinous junction

ABSTRACT: Force is transmitted between skeletal muscle and tendon via a highly specialized tissue interface: the myotendinous junction (MTJ). Despite its crucial role in mechano-transduction and involvement in muscle strain injury, knowledge about the molecular composition of the MTJ is incomplete. We performed LCMS-based proteomic analysis of human MTJ by progressive removal of the muscle component in the human semitendinosus muscle-tendon complex. Patellar tendon, which does not contains muscle tissue, was used as control. We quantified >3000 proteins across all samples and 112 proteins were significantly increased in MTJ. Of these, 24 proteins have previously been identified as MTJ proteins, including the MTJ-marker collagen XXII. Moreover, an overrepresentation of proteins related to sarcoglycan complex, laminin complex, cell junction, membrane, adhesion and extracellular matrix was observed among these proteins. We further confirmed the presence of newly identified MTJ proteins by immunofluorescence in human MTJ: Xin actin-binding repeat-containing protein 2 (XIRP2), thrombospondin-4 (THBS4), tetraspanin-24 (CD151), kindlin-2 (FERMT2), cartilage intermediate layer protein 1 (CILP) and integrin-alpha10 (ITGA10). Altogether, these data provide the first detailed proteomics resource of human MTJ.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Skeletal Muscle, Tendon

SUBMITTER: atul shahaji deshmukh

LAB HEAD: Atul S Deshmukh

PROVIDER: PXD029307 | Pride | 2022-03-01

REPOSITORIES: Pride

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Project description:This data set was downloaded from MetaboLights (http://www.ebi.ac.uk/metabolights/) accession number MTBLS322 Abstract:Reversal of fatty infiltration of pennate rotator cuff muscle after tendon release is hitherto impossible. The administration of nandrolone starting at the time of tendon release prevents the increase in fat content, but does not revert established fatty infiltration. We hypothesised that tendon release and myotendinous retraction cause alterations in lipid related gene expression leading to fatty muscle infiltration, which can be suppressed by nandrolone through its genomic actions if applied immediately after tendon release. The effects of infraspinatus tendon release and subsequent tendon repair at 16 weeks were studied in six Swiss Alpine sheep. In the interventional groups, 150mg nandrolone was administered weekly after tendon release until sacrifice (N22W, n=6) or starting at the time of repair (N6W, n=6). Infraspinatus volume, composition, expressed transcripts, lipids, and selected proteins were analyzed at baseline, 16 and 22 weeks. Tendon release reduced infraspinatus volume by 22% and increased fat content from 11% to 38%. These changes were not affected by repair. Fatty infiltration was associated with up-regulation of 227 lipid species, and increased levels of the adipocyte differentiation marker PPARG2 (peroxisome proliferator-activated receptor gamma 2). Nandrolone abrogated lipid accumulation, halved the loss in fiber area percentage, and up-regulated androgen receptor levels and transcript expression in the N22W but not the N6W group. The results document that nandrolone mitigates muscle-to-fat transformation after tendon release via a general down-regulation of lipid accumulation concomitantly with up-regulated expression of its nuclear receptor and downstream transcripts in skeletal muscle. Reduced responsiveness of retracted muscle to nandrolone as observed in the N6W group is reflected by a down-regulated transcript response.

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Novel tissue-specific markers revealed by in-depth proteome analysis of human myotendinous junction

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