Enzymatically dissociated muscle fibers display rapid dedifferentiation and impaired mitochondrial calcium control

• DOI: 10.1016/j.isci.2022.105654
• Publication Date: 2022-11-22T07:30:03Z
• Authors: Charlotte Gineste; Sonia Youhanna; Sabine U. Vorrink; Sara Henriksson; Andrés Hernández; Arthur J. Cheng; Thomas Chaillou; Andreas Buttgereit; Dominik Schneidereit; Oliver Friedrich; Kjell Hultenby; Joseph D. Bruton; Niklas Ivarsson; Linda Sandblad; Volker M. Lauschke; Håkan Westerblad
• Keywords: Cell biology; 0303 health sciences; 03 medical and health sciences; Functional aspects of cell biology; Cellular physiology; Cell- och molekylärbiologi; Science; Developmental biology; Q; Cell and Molecular Biology; Article

Cells rapidly lose their physiological phenotype upon disruption of extracellular matrix (ECM)-intracellular cytoskeleton interactions. By comparing adult mouse skeletal muscle fibers, isolated either by mechanical dissection or collagenase-induced ECM digestion, we investigated acute effects on cellular and mitochondrial morphology, transcriptomic signatures, Ca2+ handling. RNA-sequencing showed striking differences in gene expression patterns between the two isolation methods with enzymatically dissociated fibers resembling myopathic phenotypes. Mitochondrial appearance was grossly similar groups, but 3D electron microscopy revealed shorter less branched mitochondria following enzymatic dissociation. Repeated contractions resulted a prolonged accumulation which partially prevented cyclophilin inhibitors. Of importance, mice severe myopathy show pathognomonic during repeated this concealed dissociation, making an ambiguous method studies native intracellular fluxes.