bioRxiv Science⌕ Search

bioRxiv · 10.1101/280206

Polyamines support myogenesis by facilitating myoblast migration

Abstract

The regeneration of the muscle tissue relies on the differentiation of myoblasts into myocytes, to create myotubes and myofibers. Disruption of key events in this process may interfere with the correct formation or repair of muscle tissue. Polyamines, ubiquitous polycations that are essential for fundamental cellular processes, were demonstrated necessary for myogenesis; however, the mechanism by which polyamines contribute to this process has not yet been deciphered. In the present study, we examined the effect of polyamine depletion on the muscle regeneration model of C2C12 myoblasts. Our results reveal a requirement for polyamines at the very beginning of the muscle differentiation process. Myogenesis is accompanied by polyamine synthesis, even though the myoblasts contain high levels of polyamines at the moment of induction. Polyamine depletion at the time of induction, or inability to synthesize more polyamines during the first 24 hours of the process, inhibited myogenesis. Polyamine depletion inhibited the expression of all tested myogenic markers (Pax7, MyoD, Myogenin, Myf5 and Myosin heavy chain), as well as the cells migration and fusion abilities. Real time PCR analysis revealed two key early activation and migration factors, HGF and Annexin A1.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Brenner, S., Feiler, Y., Kahana, C.. 2018-03-11. Polyamines support myogenesis by facilitating myoblast migration. https://doi.org/10.1101/280206

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Role of Very Long-Chain Fatty Acyl-Ceramides in Endocytic Uptake

Specialized endoplasmic reticulum (ER) subdomains termed ER sterol exit sites (ERSES), transiently contact endocytic sites to locally supply sterols, which in turn initiate actin polymerization and drive membrane invagination. Osh2, an oxysterol-binding protein (OSBP)-related protein (ORP), mediates the non-vesicular sterol transfer at ER-endocytic contact sites (ER-ECS). In search for additional ERSES components, we identified Fat1, a fatty acid (FA) transporter and acyl-CoA synthetase with specificity for very long-chain (VLC) FAs (C20-C26). Analysis of VLC-FA anabolic and catabolic pathways revealed a specific requirement for VLC fatty acyl-ceramides (VLCA-Cers) in the constriction of endocytic invaginations. Strikingly, the Cer synthase physically interacted with the endocytic machinery, pointing to a direct role of VLCFA-Cers in membrane remodelling. Consistent with this view, molecular dynamics and live-cell imaging of a Cer probe indicated that the ORPs Osh1 and Osh3 contribute to Cer export from at ER-ECS and and that their depletion recapitulates the endocytic defects observed in the absence of VLCFA-Cer. Our findings establish that local transport of VLCA-Cers is required for endocytic budding and reveal ERSES as hubs where Cer and sterol biosynthesis and transport converge

cell biology↗

The Application of 3D Cryo-Volume Electron Microscopy to Visualize and Quantify the Photosynthetic Filamentous Cyanobacterium Anabaena

Cryogenic volume electron microscopy (cryo-vEM) is a rapidly developing nanoscale imaging approach that enables three-dimensional datasets of vitrified cells and tissues at intermediate resolutions without treatment with chemical fixatives or metal contrasting agents. Our goal was to evaluate the suitability of cryo-vEM for whole cells of photosynthetic organisms such as cyanobacteria. Two species of Anabaena, a filamentous cyanobacterium, were captured on poly-L-lysine-coated grids, vitrified by plunge-freezing into liquid ethane, and mapped by secondary electron and integrated fluorescence light microscopy to identify target cells. Cells of interest were coated with platinum, trenched with xenon, and 3D volumes collected by alternating argon milling and through-lens detector (TLD) imaging on a Thermo Scientific Hydra plasma focused ion beam scanning electron microscope. Volumes were segmented via 2.5D U-Net models, then quantified in Dragonfly 3D World. Block-face images revealed the cell wall, thylakoids, phycobilisomes, carboxysomes, cyanophycin and polyphosphate granules. Whole-cell 3D rendering allowed quantification of carboxysomes, cyanophycin granules, and measurement of thylakoid thickness and spacing in A. inaequalis and Anabaena sp. UTEX 2576. Thus, cryo-vEM provided interpretable whole-cell ultrastructure of cyanobacteria without chemical pre-treatment, and its compatibility with cryo-fluorescence and in situ cryo-ET enables contextual multi-scale perspectives from whole cells to molecular structures.

cell biology↗

KDM6A is enzymatically active at the Golgi apparatus where it is necessary for maintenance of Golgi apparatus morphology

KDM6A and KDM6B, lysine demethylases, play a central role in regulation of transcription through their demethylation activity towards H3K27me3 and have also been shown to be dynamically regulated upon epithelial-mesenchymal transition (EMT). In particular, KDM6A has been implicated in a variety of malignancies and is frequently mutated in cancers including those in kidney, bladder, colorectal, and breast. KDM6A and KDM6B function as part of the COMPASS-like protein complex (CLC) which includes methyltransferases KMT2C/KMT2D and the regulatory WRAD-subcomplex. KDM6A total protein is known to be suppressed by TGF-{beta} in MCF10A breast cells; however, our examination of protein localization by immunofluorescence and LC-MS/MS reveals that KDM6A expression is lost in the nucleus but retained in the Golgi apparatus, independent of other CLC complex members. Importantly, KDM6A isolated from a biochemically-separated subcellular membranous fraction possesses the catalytic JmjC domain and is enzymatically active, indicating that KDM6A may have non-nuclear substrates. Further, knockdown of KDM6A and treatment with TGF-{beta} similarly alter Golgi apparatus morphology, in a cell type specific manner, suggesting context-dependent regulatory functions. These findings support a model in which KDM6A is necessary to maintain Golgi apparatus morphology and is enzymatically active outside of the nucleus.

cell biology↗