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Functional dependence of neuroligin on a new non-PDZ intracellular domain

Abstract

Neuroligins, a family of postsynaptic adhesion molecules, are important in synaptogenesis through a well-characterized trans-synaptic interaction with neurexin. In addition, neuroligins are thought to drive postsynaptic assembly through binding of their intracellular domain to PSD-95. However, there is little direct evidence to support the functional necessity of the neuroligin intracellular domain in postsynaptic development. We found that presence of endogenous neuroligin obscured the study of exogenous mutated neuroligin. We therefore used chained microRNAs in rat organotypic hippocampal slices to generate a reduced background of endogenous neuroligin. On this reduced background, we found that neuroligin function was critically dependent on the cytoplasmic tail. However, this function required neither the PDZ ligand nor any other previously described cytoplasmic binding domain, but rather required a previously unknown conserved region. Mutation of a single critical residue in this region inhibited neuroligin-mediated excitatory synaptic potentiation. Finally, we found a functional distinction between neuroligins 1 and 3.

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Figure 1: Knockdown of neuroligin family necessary for functional study of cytoplasmic tail.
Figure 2: Replacement of endogenous NLGNs with wild-type and mutated NLGN3 reveals that AMPAR enhancement is dependent on a single residue in the cytoplasmic tail.
Figure 3: Critical region identified in NLGN3 is also crucial for the function of NLGN1 and NLGN4.
Figure 4: Mutation of the critical residue does not affect inhibitory synapses.
Figure 5: The effects of neuroligin on excitatory synapses are independent of excitatory synaptic activity.
Figure 6: Mechanism of synaptic enhancement by neuroligins and specific deficits of the mutant.

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Acknowledgements

We thank A.M. Craig, T. Sudhof and F. Varoqueaux for neuroligin constructs. We are grateful to K. Bjorgan for technical assistance and all members of the Nicoll laboratory and E. Dreyfuss for discussion of and comments on the manuscript. This work was supported by grants from the US National Institute of Mental Health.

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E.S. initiated the project and generated preliminary data. S.L.S. designed experiments, performed all electrophysiology and all imaging in slice, constructed all new constructs, produced virus, conducted data analysis and wrote the manuscript. T.H. and B.-S.C. performed all biochemical experiments. T.H. designed and carried out imaging in dissociated neurons. R.A.N. and K.W.R. supervised the project. R.A.N., K.W.R. and E.S. provided comments on the manuscript.

Corresponding author

Correspondence to Roger A Nicoll.

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The authors declare no competing financial interests.

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Shipman, S., Schnell, E., Hirai, T. et al. Functional dependence of neuroligin on a new non-PDZ intracellular domain. Nat Neurosci 14, 718–726 (2011). https://doi.org/10.1038/nn.2825

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