title: Neural cell-cell and cell-substrate adhesion through N-cadherin, N-CAM and L1.
abstract: In this study neural (N)-cadherin, neural cell adhesion molecule (N-CAM) and L1 proteins and their antibody equivalents were covalently immobilized on a polyethylene-imine (PEI)-coated glass surface to form neuron-adhesive coatings. Impedance sensing and (supplementary) image analysis were used to monitor the effects of these CAMs. Immobilization of high concentrations of both N-cadherin protein and antibody led to good adhesion of neurons to the modified surface, better than surfaces treated with 30.0 and 100.0 µg ml(-1) N-CAM protein and antibody. L1 antibody and protein coating revealed no significant effect on neuronal cell-substrate adhesion. In a second series of combinatorial experiments, we used the same antibodies and proteins as medium-additives to inhibit cell-cell adhesion between neurons. Adhesion of neurons cultured on N-cadherin protein or antibody-modified surfaces was lowered by the addition of a soluble N-cadherin protein and antibody to the culturing medium, accelerating neuronal aggregation. The presence of a soluble N-CAM antibody or protein had no effect on the adhesion of neuronal cells on a N-cadherin protein-modified surface. On a N-cadherin antibody-coated surface, the addition of a soluble N-CAM protein led to cell death of neurons after 48 h, while a N-CAM antibody had no effect. In the presence of a soluble N-cadherin protein and antibody the aggregation of neurons was inhibited, both on N-CAM protein and N-CAM antibody-modified surfaces. Neurons cultured on immobilized antibodies were less affected by the addition of soluble CAM blockers than neurons cultured on immobilized proteins, indicating that antibody-protein bonds are more stable compared to protein-protein bonds.
authors.affiliation: Neural and Cellular Engineering, MIRA Institute, University of Twente, Enschede, The Netherlands. remywiertz@hotmail.com
authors.forename: R W F
authors.initials: RW
authors.lastname: Wiertz
authors.forename: E
authors.initials: E
authors.lastname: Marani
authors.forename: W L C
authors.initials: WL
authors.lastname: Rutten
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D019138:Antibodies, Blocking / Q000494:pharmacology; D015820:Cadherins / Q000037:antagonists & inhibitors / Q000276:immunology / Q000502:physiology*; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D017097:Electric Impedance; D004566:Electrodes; D039841:Leukocyte L1 Antigen Complex / Q000276:immunology / Q000502:physiology*; D019006:Neural Cell Adhesion Molecules / Q000037:antagonists & inhibitors / Q000276:immunology / Q000502:physiology*; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D051381:Rats; D013499:Surface Properties
chemical_list: D019138:Antibodies, Blocking; D015820:Cadherins; D039841:Leukocyte L1 Antigen Complex; D019006:Neural Cell Adhesion Molecules