GREG cells, a dysferlin-deficient myogenic mouse cell line.
The dysferlinopathies (e.g. LGMD2b, Myoshi myopathy) are progressive, adult-onset muscle wasting syndromes caused by mutations in the gene coding for dysferlin. Dysferlin is a large (~200kDa) membrane-anchored protein, required for maintenance of plasmalemmal integrity in muscle fibers. To facilitate analysis of dysferlin function in muscle cells, we have established a dysferlin-deficient myogenic cell line (GREG cells) from the A/J mouse, a genetic model for dysferlinopathy. GREG cells have no detectable dysferlin expression, but proliferate normally in growth medium and fuse into functional myotubes in differentiation medium. GREG myotubes exhibit deficiencies in plasma membrane repair, as measured by laser wounding in the presence of FM1-43 dye. Under the wounding conditions used, the majority (~66%) of GREG myotubes lack membrane repair capacity, while no membrane repair deficiency was observed in dysferlin-normal C2C12 myotubes, assayed under the same conditions. We discuss the possibility that the observed heterogeneity in membrane resealing represents genetic compensation for dysferlin deficiency.
Program in Physical Biology, Eunice Kennedy Schriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA.
Glen W
GW
Humphrey
Elena
E
Mekhedov
Paul S
PS
Blank
Antoine
A
de Morree
Gulcin
G
Pekkurnaz
Kanneboyina
K
Nagaraju
Joshua
J
Zimmerberg
D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D000073939:Dysferlin; D007834:Lasers / Q000009:adverse effects; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D024510:Muscle Development / Q000235:genetics; D018485:Muscle Fibers, Skeletal / Q000378:metabolism; D018482:Muscle, Skeletal / Q000293:injuries / Q000378:metabolism; D049288:Muscular Dystrophies, Limb-Girdle / Q000235:genetics*; D032446:Myoblasts / Q000378:metabolism*; D011726:Pyridinium Compounds / Q000009:adverse effects; D000644:Quaternary Ammonium Compounds / Q000009:adverse effects; D014947:Wounds and Injuries / Q000235:genetics
C497505:Dysf protein, mouse; D000073939:Dysferlin; C073804:FM1 43; D008565:Membrane Proteins; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds