Membrane repair of human skeletal muscle cells requires Annexin-A5.
Defect in membrane repair contributes to the development of limb girdle muscular dystrophy type 2B (LGMD2B) and Miyoshi myopathy. In healthy skeletal muscle, unraveling membrane repair mechanisms requires to establish an exhaustive list of the components of the resealing machinery. Here we show that human myotubes rendered deficient for Annexin-A5 (AnxA5) suffer from a severe defect in membrane resealing. This defect is rescued by the addition of recombinant AnxA5 while an AnxA5 mutant, which is unable to form 2D protein arrays, has no effect. Using correlative light and electron microscopy, we show that AnxA5 binds to the edges of the torn membrane, as early as a few seconds after sarcolemma injury, where it probably self-assembles into 2D arrays. In addition, we observed that membrane resealing is associated with the presence of a cluster of lipid vesicles at the wounded site. AnxA5 is present at the surface of these vesicles and may thus participate in plugging the cell membrane disruption. Finally, we show that AnxA5 behaves similarly in myotubes from a muscle cell line established from a patient suffering from LGMD2B, a myopathy due to dysferlin mutations, which indicates that trafficking of AnxA5 during sarcolemma damage is independent of the presence of dysferlin.
Institute of Chemistry and Biology of Membranes and Nano-objects, UMR 5248, CNRS, University of Bordeaux, IPB, F-33600 Pessac, France.
Romain
R
Carmeille
Institute of Chemistry and Biology of Membranes and Nano-objects, UMR 5248, CNRS, University of Bordeaux, IPB, F-33600 Pessac, France.
Flora
F
Bouvet
Institute of Chemistry and Biology of Membranes and Nano-objects, UMR 5248, CNRS, University of Bordeaux, IPB, F-33600 Pessac, France.
Sisareuth
S
Tan
Institute of Chemistry and Biology of Membranes and Nano-objects, UMR 5248, CNRS, University of Bordeaux, IPB, F-33600 Pessac, France.
Coralie
C
Croissant
Institute of Chemistry and Biology of Membranes and Nano-objects, UMR 5248, CNRS, University of Bordeaux, IPB, F-33600 Pessac, France.
Céline
C
Gounou
Sorbonne Universités, UPMC Univ Paris 06, Inserm UMRS974, CNRS FRE3617, Center for Research in Myology, 47 Boulevard de l'hôpital, 75013 Paris, France.
Kamel
K
Mamchaoui
Sorbonne Universités, UPMC Univ Paris 06, Inserm UMRS974, CNRS FRE3617, Center for Research in Myology, 47 Boulevard de l'hôpital, 75013 Paris, France.
Vincent
V
Mouly
Institute of Chemistry and Biology of Membranes and Nano-objects, UMR 5248, CNRS, University of Bordeaux, IPB, F-33600 Pessac, France.
Alain R
AR
Brisson
Institute of Chemistry and Biology of Membranes and Nano-objects, UMR 5248, CNRS, University of Bordeaux, IPB, F-33600 Pessac, France. Electronic address: a.bouter@cbmn.u-bordeaux.fr.
Anthony
A
Bouter
D000328:Adult; D017304:Annexin A5 / Q000378:metabolism* / Q000648:ultrastructure; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D000073939:Dysferlin; D005110:Extracellular Space / Q000378:metabolism; D006801:Humans; D007834:Lasers; D008051:Lipid Bilayers / Q000378:metabolism; D008565:Membrane Proteins / Q000172:deficiency / Q000378:metabolism; D018485:Muscle Fibers, Skeletal / Q000378:metabolism* / Q000473:pathology* / Q000648:ultrastructure; D009124:Muscle Proteins / Q000172:deficiency / Q000378:metabolism; D049288:Muscular Dystrophies, Limb-Girdle / Q000378:metabolism / Q000473:pathology; D009154:Mutation / Q000235:genetics; D032446:Myoblasts / Q000378:metabolism / Q000473:pathology; D011994:Recombinant Proteins / Q000378:metabolism; D012508:Sarcolemma / Q000473:pathology; D013347:Subcellular Fractions / Q000378:metabolism; D014945:Wound Healing
D017304:Annexin A5; C114839:DYSF protein, human; D000073939:Dysferlin; D008051:Lipid Bilayers; D008565:Membrane Proteins; D009124:Muscle Proteins; D011994:Recombinant Proteins
Annexin A5; Electron microscopy; Laser injury; Limb girdle muscular dystrophy; Membrane repair; Skeletal muscle