title: Assembly of synapses: biomimetic assays to control neurexin/neuroligin interactions at the neuronal surface.
abstract: The role of adhesion molecules in the assembly of synapses in the nervous system is an important issue. To characterize the role of neurexin/neuroligin adhesion complexes in synapse differentiation, various imaging assays can be performed in primary hippocampal cultures. First, to temporally control contact formation, biomimetic assays can be performed using microspheres coated with purified neurexin or with antibody clusters that aggregate neurexin. These models are combined with live fluorescence imaging to study the dynamics of accumulation of post-synaptic components, including scaffolding molecules and glutamate receptors. To demonstrate that AMPA receptors can be recruited to nascent neurexin/neuroligin contacts through lateral diffusion, the mobility of AMPA receptors in the neuronal membrane is monitored by tracking individual quantum dots (QDs) conjugated to antibodies against AMPA receptors. Experiments monitoring the attachment and detachment of Nrx-coated QDs to measure the rates of neurexin/neuroligin interaction can also be performed. Each of these assays is detailed in this unit.
authors.affiliation: Université Bordeaux, Bordeaux Imaging Center, Bordeaux, France.
authors.forename: Magali
authors.initials: M
authors.lastname: Mondin
authors.forename: Béatrice
authors.initials: B
authors.lastname: Tessier
authors.forename: Olivier
authors.initials: O
authors.lastname: Thoumine
mesh_terms: D000818:Animals; D032701:Biomimetics / Q000379:methods*; D002135:Calcium-Binding Proteins; D015816:Cell Adhesion Molecules, Neuronal / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D019006:Neural Cell Adhesion Molecules; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology; D025941:Protein Interaction Mapping / Q000379:methods; D060066:Protein Interaction Maps; D051381:Rats; D013569:Synapses / Q000378:metabolism*
chemical_list: D002135:Calcium-Binding Proteins; D015816:Cell Adhesion Molecules, Neuronal; D008565:Membrane Proteins; C539628:NRXN1 protein, human; D009419:Nerve Tissue Proteins; D019006:Neural Cell Adhesion Molecules; C093438:neuroligin 1