ABSTRACT
Biomimetic membranes create opportunities for various applications, including the possibility of replacing interacting cells in a cell-cell contact. Here we have fractionated synthetic membranes using metal nano-grid structures where EphrinA5 (EA5), a neuronal adhesion promoter, was anchored via its Fc domain (immunoglobulin G (IgG)-domain). FRAP experiments were performed to check the confinement of the synthetic membrane within these nano-structures. Rat cortical primary neurons were cultured and live cell imaging techniques were used to monitor the neuronal interaction with these fractionated synthetic membranes. Computational imaging analysis of the corresponding images elucidated interesting details of the cellular behavior. The phenotypic cellular response on these nano-membrane fractions was found to be similar to that on non-fractionated synthetic membranes indicating that although the number of focal adhesion points was low (due to the reduced EA5 number) in the nano-sized membrane patches perhaps some other factors like metal grid boundaries might be playing a role in rendering the similarity.
Subject(s)
Cell Adhesion , Ephrin-A5/chemistry , Membranes, Artificial , Nanostructures , Neurons/cytology , Animals , Cells, Cultured , Female , Immunoglobulin G , Rats , Rats, WistarABSTRACT
Artificial lipid bilayer on solid substrate plays an important role as an interface between nanotechnology and biology. In this study, grid structures were patterned on Au-Nb-glass substrate and artificial bilayer was prepared on these structures. The fluidity was checked using fluorescence recovery after photobleaching (FRAP), and neuronal adhesion was monitored on such structure using EphrinA5-tethered lipid bilayer. EphrinA5 is a ligand that binds to the Eph receptors of rat cortical neurons and influences cellular adhesion. Our result elucidated that influence of these nanopatterned protein-tethered lipid bilayer on cellular guidance and signaling can address many underlying mechanisms of cellular functioning and help us to understand and differentiate the signaling procedure in cancer and neurodegenerative diseases.