Dissection of synapse induction by neuroligins: effect of a neuroligin mutation associated with autism.
To study synapse formation by neuroligins, we co-cultured hippocampal neurons with COS cells expressing wild type and mutant neuroligins. The large size of COS cells makes it possible to test the effect of neuroligins presented over an extended surface area. We found that a uniform lawn of wild type neuroligins displayed on the cell surface triggers the formation of hundreds of uniformly sized, individual synaptic contacts that are labeled with neurexin antibodies. Electron microscopy revealed that these artificial synapses contain a presynaptic active zone with docked vesicles and often feature a postsynaptic density. Neuroligins 1, 2, and 3 were active in this assay. Mutations in two surface loops of neuroligin 1 abolished neuroligin binding to neurexin 1beta, a presumptive presynaptic binding partner for postsynaptic neuroligins, and blocked synapse formation. An analysis of mutant neuroligins with an amino acid substitution that corresponds to a mutation described in patients with an autistic syndrome confirmed previous reports that these mutant neuroligins have a compromised capacity to be transported to the cell surface. Nevertheless, the small percentage of mutant neuroligins that reached the cell surface still induced synapse formation. Viewed together, our data suggest that neuroligins generally promote artificial synapse formation in a manner that is associated with beta-neurexin binding and results in morphologically well differentiated synapses and that a neuroligin mutation found in autism spectrum disorders impairs cell-surface transport but does not completely abolish synapse formation activity.
Chubykin
Alexander A
AA
Center for Basic Neuroscience, Department of Molecular Genetics, and Howard Hughes Medical Institute, The University of Texas Southwestern Medical Center at Dallas, 6000 Harry Hines Boulevard, Dallas, TX 75390-9111, USA.
Liu
Xinran
X
Comoletti
Davide
D
Tsigelny
Igor
I
Taylor
Palmer
P
Südhof
Thomas C
TC
D000818:Animals; D001321:Autistic Disorder / Q000235:genetics*; D019556:COS Cells; D015816:Cell Adhesion Molecules, Neuronal; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D018920:Coculture Techniques; D005822:Genetic Vectors; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D013569:Synapses / Q000378:metabolism*; D014162:Transfection
D015816:Cell Adhesion Molecules, Neuronal; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; C093438:neuroligin 1; C098003:neuroligin 2; C098004:neuroligin 3; C086959:neurexin Ibeta