title: Biochemical and biophysical changes underlie the mechanisms of basement membrane disruptions in a mouse model of dystroglycanopathy.
abstract: Mutations in glycosyltransferases, such as protein O-mannose N-acetylglucosaminyltransferase 1 (POMGnT1), causes disruptions of basement membranes (BMs) that results in neuronal ectopias and muscular dystrophy. While the mutations diminish dystroglycan-mediated cell-ECM interactions, the cause and mechanism of BM disruptions remain unclear. In this study, we established an in vitro model to measure BM assembly on the surface of neural stem cells. Compared to control cells, the rate of BM assembly on POMGnT1 knockout neural stem cells was significantly reduced. Further, immunofluorescence staining and quantitative proteomic analysis of the inner limiting membrane (ILM), a BM of the retina, revealed that laminin-111 and nidogen-1 were reduced in POMGnT1 knockout mice. Finally, atomic force microscopy showed that the ILM from POMGnT1 knockout mice was thinner with an altered surface topography. The results combined demonstrate that reduced levels of key BM components cause physical changes that weaken the BM in POMGnT1 knockout mice. These changes are caused by a reduced rate of BM assembly during the developmental expansion of the neural tissue.
authors: Department of Neuroscience and Physiology, SUNY Upstate Medical University, USA.
authors: Peng
authors: P
authors: Zhang
authors: Yuan
authors: Y
authors: Yang
authors: Joseph
authors: J
authors: Candiello
authors: Trista L
authors: TL
authors: Thorn
authors: Noel
authors: N
authors: Gray
authors: Willi M
authors: WM
authors: Halfter
authors: Huaiyu
authors: H
authors: Hu
mesh_terms: D000818:Animals; D001485:Basement Membrane / Q000378:metabolism / Q000473:pathology*; D002478:Cells, Cultured; D049030:Dystroglycans / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D006031:Glycosylation; D007797:Laminin / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D009136:Muscular Dystrophies / Q000378:metabolism / Q000473:pathology*; D017351:N-Acetylglucosaminyltransferases / Q000172:deficiency / Q000235:genetics*; D058953:Neural Stem Cells / Q000378:metabolism; D009457:Neuroglia / Q000378:metabolism; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D012160:Retina / Q000378:metabolism / Q000473:pathology; D018874:Spheroids, Cellular / Q000378:metabolism
chemical_list: D016326:Extracellular Matrix Proteins; D007797:Laminin; D008562:Membrane Glycoproteins; C029967:nidogen; D049030:Dystroglycans; D017351:N-Acetylglucosaminyltransferases; C443106:protein O-mannose beta-1,2-N-acetylglucosaminyltransferase