title: Neurexin Ibeta and neuroligin are localized on opposite membranes in mature central synapses.
abstract: Synaptogenesis requires formation of trans-synaptic complexes between neuronal cell-adhesion receptors. Heterophilic receptor pairs, such as neurexin Ibeta and neuroligin, can mediate distinct intracellular signals and form different cytoplasmic scaffolds in the pre- and post-synaptic neuron, and may be particularly important for synaptogenesis. However, the functions of neurexin and neuroligin depend on their distribution in the synapse. Neuroligin has been experimentally assigned to the post-synaptic membrane, while the localization of neurexin remains unclear. To study the subcellular distribution of neurexin Ibeta and neuroligin in mature cerebrocortical synapses, we have developed a novel method for the physical separation of junctional membranes and their direct analysis by western blotting. Using urea and dithiothreitol, we disrupted trans-synaptic protein links, without dissolving the lipid phase, and fractionated the pre- and post-synaptic membranes. The purity of these fractions was validated by electron microscopy and western blotting using multiple synaptic markers. A quantitative analysis has confirmed that neuroligin is localized strictly in the post-synaptic membrane. We have also demonstrated that neurexin Ibeta is largely (96%) pre-synaptic. Thus, neurexin Ibeta and neuroligin normally form trans-synaptic complexes and can transduce bidirectional signals.
authors.affiliation: Division of Cell and Molecular Biology, Imperial College London, London, UK.
authors.forename: Otto
authors.initials: O
authors.lastname: Berninghausen
authors.forename: M Atiqur
authors.initials: MA
authors.lastname: Rahman
authors.forename: John-Paul
authors.initials: JP
authors.lastname: Silva
authors.forename: Bazbek
authors.initials: B
authors.lastname: Davletov
authors.forename: Colin
authors.initials: C
authors.lastname: Hopkins
authors.forename: Yuri A
authors.initials: YA
authors.lastname: Ushkaryov
mesh_terms: D000818:Animals; D015816:Cell Adhesion Molecules, Neuronal; D002462:Cell Membrane / Q000378:metabolism*; D002540:Cerebral Cortex / Q000166:cytology*; D008565:Membrane Proteins / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission / Q000379:methods; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013347:Subcellular Fractions / Q000378:metabolism; D013569:Synapses / Q000378:metabolism / Q000648:ultrastructure*; D013572:Synaptic Vesicles / Q000378:metabolism*
chemical_list: D015816:Cell Adhesion Molecules, Neuronal; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; C093438:neuroligin 1; C086959:neurexin Ibeta