Article Text

Download PDFPDF

11 The role of N-glycosylation of the NOTCH1 receptor in jagged1-stimulated myogenic differentiation in vitro
  1. Abidemi Olayinka,
  2. Eoin Corcoran,
  3. Brendan O’Connor,
  4. Paul A Cahill
  1. Dublin City University, Vascular Biology and Therapeutics Group, School of Biotechnology


Resident stem cell fate decisions within the vasculature are crucial to the pathogenesis of vascular diseases, including, arteriosclerosis, atherosclerosis and in-stent restenosis after angioplasty. The Notch signalling pathway regulates stem cell fate and is highly regulated by a number of mechanisms including glycosylation, a post-translational modification.

Our main objective was to define a putative role for N-glycosylation of Notch1 receptor in controlling resident vascular stem cell fate in vitro. Utilising ligand-induced Notch signalling assay with Jagged1, qRT-PCR, immunocytochemistry, ectopic expression of Notch1 receptor, siRNA knockdown, pharmacological inhibition and enzyme linked lectin assay (ELLA), alterations in N-glycan decoration of the Notch1 receptor were assessed before evaluation of their effects on Notch signalling and Notch ligand promotion of myogenic differentiation.

N-glycosylation of the Notch1 receptor was assessed using a combination of the HPLC and ELLA assays and confirmed the presence of N-glycans on the receptor, an effect that was abrogated following inhibition of glycosyltransferase activity with tunicamycin and lunatic fringe (Lfng) knockdown. Jagged1- induced Notch activation increased Notch target gene expression and promoted myogenic differentiation of bone-marrow derived mesenchymal stem cells and resident vascular stem cells. Selective knockdown of the Notch1 receptor in stem cells resulted in a significant decrease in Jagged1 stimulated Hey1 expression, a Notch1 target gene, concomitant with a reduction in myogenic differentiation due to decreased smooth muscle differentiation marker expression (CNN1 and MYH11 mRNA and protein levels). Inhibition of N-glycosylation with tunicamycin lead to a down regulation of smooth muscle differentiation markers, CNN1 and MYH11 independent of a reduction in Notch target gene expression. Lfng knockdown lead to a similar significant reduction in Jagged1 induced myogenic differentiation (reduced CNN1 expression). Collectively, these results suggest that N-glycosylation of Notch1 receptor is involved in Notch signalling leading to altered resident vascular stem cell fate.

Statistics from

Request Permissions

If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.