We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
N-glycosylation inhibition impairs C2C12 and L6 myoblast differentiation and IGF-1 signalling.
- Authors
Annibalini, Giosuè; Bocconcelli, Matteo; Saltarelli, Roberta; Valli, Giacomo; Barberi, Laura; Musarò, Antonio; Barbieri, Elena
- Abstract
Several muscular diseases are associated with aberrant protein glycosylation, suggesting essential glycanmediated functions in myogenesis and muscle development (1). However, the role played by protein Nglycosylation in the process of muscle differentiation remains poorly characterized. Here, we used C2C12 and L6 muscle cell cultures to investigate the effects of pharmacological inhibition of N-glycosylation by Tunicamycin (TUN) on myoblast differentiation. Non-toxic doses of TUN (0.01µg/ml) inhibited C2C12 and L6 myoblast fusion and disrupted the coordinated temporal expression of myogenic regulator genes Ccnd1, MyoD, Myogenin and Mrf4. C2C12 control myotubes also showed an increase lectin binding (ConA; PHA-L and AAL) compared to myoblasts, while lectin reactivities decreased in TUN-treated myotubes indicating a N-glycosylation deficiency. Intrestingly, similar resulst were obtained by genetic knockdown of phosphomannomutase 2 (PMM2) gene in C2C12 cells, which encode an enzyme essential for catalysing an early step of the N-glycosylation pathway (2). Finally, TUN treatment decreased the IGF-1R level and markedly attenuated the IGF-1-induced ERK-1/2 and Akt phosphorylation (3). These results suggest that impaired myoblast differentiation could be a key factor in the pathophysiology of muscle-related manifestations commonly found in congenital disorders of Nglycosylation. Our data form a valuable resource to further understand the glycobiology of myogenesis and will aid to explain the association between abnormal Nglycosylation and defects in muscle development and regeneration commonly found in individuals with Congenital disorders of glycosylation (CDG), such as PMM2-CDG, and other disease associated with protein hypo-Nglycosylation. Acronyms PMM2, Phosphomannomutase 2; Ccnd1, Cyclin D1; PCNA, Proliferating Cell Nuclear Antigen; MyoD, Myogenic differentiation 1; Mrf4, Myogenic Factor 6; Con A, Concanavalin A; PHA-L, Phaseolus vulgaris leucoagglutinin; AAL, Aleuria Aurantia (AAL); IGF-1R, Insulin-like growth factor-1 receptor; AKT, Serine/threonine protein kinase B; ERK1/2 Extracellular signal-regulated kinase 1/2.
- Subjects
PROLIFERATING cell nuclear antigen; CONCANAVALIN A; PROTEIN kinase B; MUSCLE growth; REGULATOR genes
- Publication
European Journal of Translational Myology, 2023, Vol 33, Issue 2, p44
- ISSN
2037-7452
- Publication type
Article
- DOI
10.4081/ejtm.2023.11427