The synaptic CT carbohydrate modulates binding and expression of extracellular matrix proteins in skeletal muscle: Partial dependence on utrophin.
The CT carbohydrate, Neu5Ac/Neu5Gcalpha2,3[GalNAcbeta1,4]Galbeta1,4GlcNAcbeta-, is specifically expressed at the neuromuscular junction in skeletal myofibers of adult vertebrates. When Galgt2, the glycosyltransferase that creates the synaptic beta1,4GalNAc portion of this glycan, is overexpressed in extrasynaptic regions of the myofiber membrane, alpha dystroglycan becomes glycosylated with the CT carbohydrate and this coincides with the ectopic expression of synaptic dystroglycan-binding proteins, including laminin alpha4, laminin alpha5, and utrophin. Here we show that both synaptic and extrasynaptic forms of laminin and agrin have increased binding to the CT carbohydrate compared to sialyl-N-acetyllactosamine, its extrasynaptically expressed precursor. Muscle laminins also show increased binding to CT-glycosylated muscle alpha dystroglycan relative to its non-CT-containing glycoforms. Overexpression of Galgt2 in transgenic mouse skeletal muscle increased the mRNA expression of extracellular matrix (ECM) genes, including agrin and laminin alpha5, as well as utrophin, integrin alpha7, and neuregulin. Increased expression of ECM proteins in Galgt2 transgenic skeletal muscles was partially dependent on utrophin, but utrophin was not required for Galgt2-induced changes in muscle growth or neuromuscular development. These experiments demonstrate that overexpression of a synaptic carbohydrate can increase both ECM binding to alpha dystroglycan and ECM expression in skeletal muscle, and they suggest a mechanism by which Galgt2 overexpression may inhibit muscular dystrophy and affect neuromuscular development.
Yoon
Jung Hae
JH
Center for Gene Therapy, The Research Institute at Nationwide Children's Hospital, Department of Pediatrics, Ohio State University College of Medicine, 700 Children's Drive, Columbus, OH 43205, USA.
Chandrasekharan
Kumaran
K
Xu
Rui
R
Glass
Matthew
M
Singhal
Neha
N
Martin
Paul T
PT
D018171:Agrin / Q000378:metabolism; D000818:Animals; D002461:Cell Line, Transformed; D049030:Dystroglycans / Q000378:metabolism; D016189:Dystrophin / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology; D016695:Glycosyltransferases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D034882:Lamins / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D018482:Muscle, Skeletal / Q000378:metabolism*; D009469:Neuromuscular Junction / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects / Q000235:genetics; D014162:Transfection / Q000379:methods; D049411:Utrophin / Q000235:genetics / Q000378:metabolism*
D018171:Agrin; D016189:Dystrophin; D016326:Extracellular Matrix Proteins; D034882:Lamins; D049411:Utrophin; D049030:Dystroglycans; D016695:Glycosyltransferases; C505903:Galgt2 protein, mouse