The function of neurofascin155 in oligodendrocytes is regulated by metalloprotease-mediated cleavage and ectodomain shedding.
Formation of the paranodal axo-glial junction requires the oligodendrocyte-specific 155-kDa isoform of neurofascin (NF155). Here, we report the presence of two peptides in cultured oligodendrocytes, which are recognized by distinct NF155-specific antibodies and correspond to a membrane anchor of 30 kDa and a 125 kDa peptide, which is shed from the cells, indicating that it consists of the NF155 ectodomain. Transfection of OLN-93 cells with NF155 verified that both peptides originate from NF155 cleavage, and we present evidence that metalloproteases mediate NF155 processing. Interestingly, metalloprotease activity is required for NF155 transport into oligodendrocyte processes supporting the functional significance of NF155 cleavage. To further characterize NF155 cleavage and function, we transfected MDCK cells with NF155. Although ectodomain shedding was observed in polarized and non-polarized MDCK cells, surface localization of NF155 was restricted to the lateral membrane of polarized cells consistent with a role in cell-cell adhesion. Aggregation assays performed with OLN-93 cells confirmed that NF155 accelerates cell-cell adhesion in a metalloprotease-dependent manner. The physiological relevance of NF155 processing is corroborated by the presence of NF155 cleavage products in heavy myelin, suggesting a role of NF155 ectodomain shedding for the generation and/or stabilization of the nodal/paranodal architecture.
Maier
Olaf
O
University Medical Center Groningen, University of Groningen, Department of Cell Biology/Section Membrane Cell Biology, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands. o.maier@med.umcg.nl
van der Heide
Tiemen
T
Johnson
Richard
R
de Vries
Hans
H
Baron
Wia
W
Hoekstra
Dick
D
D000818:Animals; D000831:Animals, Newborn; D001923:Brain Chemistry; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D015815:Cell Adhesion Molecules / Q000378:metabolism / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology; D049108:Cell Enlargement / Q000187:drug effects; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D002478:Cells, Cultured; D004151:Dipeptides / Q000494:pharmacology; D004847:Epithelial Cells / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D005260:Female; D045726:Metalloproteases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009186:Myelin Sheath / Q000737:chemistry; D009414:Nerve Growth Factors / Q000378:metabolism / Q000502:physiology*; D009836:Oligodendroglia / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D020033:Protein Isoforms / Q000378:metabolism / Q000502:physiology; D011499:Protein Processing, Post-Translational; D051381:Rats; D017208:Rats, Wistar; D014162:Transfection
D015815:Cell Adhesion Molecules; D004151:Dipeptides; C078131:N-(2(R)-2-(hydroxamidocarbonylmethyl)-4-methylpentanoyl)-L-tryptophan methylamide; D009414:Nerve Growth Factors; C504594:Nfasc protein, rat; D011480:Protease Inhibitors; D020033:Protein Isoforms; D045726:Metalloproteases