Reduced proliferative activity of primary POMGnT1-null myoblasts in vitro.
Protein O-linked mannose beta1,2-N-acetylglucosaminyltransferase 1 (POMGnT1) is an enzyme that transfers N-acetylglucosamine to O-mannose of glycoproteins. Mutations of the POMGnT1 gene cause muscle-eye-brain (MEB) disease. To obtain a better understanding of the pathogenesis of MEB disease, we mutated the POMGnT1 gene in mice using a targeting technique. The mutant muscle showed aberrant glycosylation of alpha-DG, and alpha-DG from mutant muscle failed to bind laminin in a binding assay. POMGnT1(-/-) muscle showed minimal pathological changes with very low-serum creatine kinase levels, and had normally formed muscle basal lamina, but showed reduced muscle mass, reduced numbers of muscle fibers, and impaired muscle regeneration. Importantly, POMGnT1(-/-) satellite cells proliferated slowly, but efficiently differentiated into multinuclear myotubes in vitro. Transfer of a retrovirus vector-mediated POMGnT1 gene into POMGnT1(-/-) myoblasts completely restored the glycosylation of alpha-DG, but proliferation of the cells was not improved. Our results suggest that proper glycosylation of alpha-DG is important for maintenance of the proliferative activity of satellite cells in vivo.
Miyagoe-Suzuki
Yuko
Y
Department of Molecular Therapy, National Institute of Neuroscience, National Center of Neurology and Psychiatry, 4-1-1 Ogawahigashi, Kodaira, Tokyo 187-8502, Japan. miyagoe@ncnp.go.jp
Masubuchi
Nami
N
Miyamoto
Kaori
K
Wada
Michiko R
MR
Yuasa
Shigeki
S
Saito
Fumiaki
F
Matsumura
Kiichiro
K
Kanesaki
Hironori
H
Kudo
Akira
A
Manya
Hiroshi
H
Endo
Tamao
T
Takeda
Shin'ichi
S
D000818:Animals; D049109:Cell Proliferation; D002478:Cells, Cultured; D003402:Creatine Kinase / Q000097:blood; D053595:Embryonic Stem Cells / Q000378:metabolism; D005355:Fibrosis / Q000150:complications / Q000201:enzymology / Q000473:pathology; D017353:Gene Deletion; D018390:Gene Targeting; D007150:Immunohistochemistry; D051379:Mice; D018482:Muscle, Skeletal / Q000201:enzymology / Q000473:pathology / Q000648:ultrastructure; D009137:Muscular Dystrophy, Animal / Q000097:blood / Q000150:complications / Q000201:enzymology / Q000473:pathology; D032446:Myoblasts / Q000166:cytology* / Q000201:enzymology* / Q000648:ultrastructure; D017351:N-Acetylglucosaminyltransferases / Q000172:deficiency* / Q000378:metabolism; D010641:Phenotype; D012038:Regeneration; D032496:Satellite Cells, Skeletal Muscle / Q000201:enzymology / Q000473:pathology / Q000648:ultrastructure; D015398:Signal Transduction
D017351:N-Acetylglucosaminyltransferases; C443106:protein O-mannose beta-1,2-N-acetylglucosaminyltransferase; D003402:Creatine Kinase