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PIP3 controls synaptic function by maintaining AMPA receptor clustering at the postsynaptic membrane

Abstract

Despite their low abundance, phosphoinositides are critical regulators of intracellular signaling and membrane compartmentalization. However, little is known of phosphoinositide function at the postsynaptic membrane. Here we show that continuous synthesis and availability of phosphatidylinositol-(3,4,5)-trisphosphate (PIP3) at the postsynaptic terminal is necessary for sustaining synaptic function in rat hippocampal neurons. This requirement was specific for synaptic, but not extrasynaptic, AMPA receptors, nor for NMDA receptors. PIP3 downregulation impaired PSD-95 accumulation in spines. Concomitantly, AMPA receptors became more mobile and migrated from the postsynaptic density toward the perisynaptic membrane within the spine, leading to synaptic depression. Notably, these effects were only revealed after prolonged inhibition of PIP3 synthesis or by direct quenching of this phosphoinositide at the postsynaptic cell. Therefore, we conclude that a slow, but constant, turnover of PIP3 at synapses is required for maintaining AMPA receptor clustering and synaptic strength under basal conditions.

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Figure 1: Expression of PH-GRP1 in hippocampal neurons and specific binding to PIP3.
Figure 2: Bidirectional modulation of AMPA-receptor mediated currents by PIP3.
Figure 3: Inhibition of PIP3 synthesis produces a slow and gradual depression of AMPA receptor-mediated transmission.
Figure 4: PIP3 is required for LTP and affects both constitutively cycling and regulated populations of AMPA receptors.
Figure 5: Depletion of PIP3 leads to the accumulation of AMPA receptors in dendritic spines.
Figure 6: Depletion of PIP3 impairs PSD-95 accumulation in spines and increases surface mobility of GluA2 recombinant receptors.
Figure 7: Depletion of PIP3 causes a redistribution of AMPARs between the PSD and extrasynaptic membrane.
Figure 8: Inhibition of glutamate reuptake abolishes PH-GRP1-induced depression of AMPA receptor-mediated transmission.

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Acknowledgements

We thank R. Holz (University of Michigan) for the plasmid containing the PH-PLC sequence, A. Lee (University of Michigan) for the plasmid containing the catalytic domain of mouse PI3K (p110α), C. Dotti (Katholieke Universiteit Leuven) for the membrane-anchored GFP-CAAX construct, R. Malinow (University of California, San Diego) for the SEP-GluA2 construct and L. Chen (University of California, Berkeley) for facilitating some of the experiments carried out by K.L.A. We also thank S. Jurado and members of the Esteban laboratory for critical reading of this manuscript, and S. Fisher, E. Stuenkel, G. Murphy and R. Holz for discussions. This work was supported by grants from the US National Institute of Mental Health (J.A.E. and J.R.M.), the Dana Foundation (J.A.E.) and the Spanish Ministry of Science and Innovation (J.A.E.). M.F.-M. and S.K. are supported by postdoctoral contracts, and M.R. by a predoctoral fellowship, from the Spanish Ministry of Science and Innovation.

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K.L.A. is responsible for most of the experimental work. M.R., M.F.-M. and S.K. contributed some of the biochemical and imaging experiments. C.N.P. carried out cloning and provided technical support. J.R.M. designed and supervised some of the experiments. K.L.A. and J.A.E. designed the experiments and wrote the manuscript.

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Correspondence to José A Esteban.

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Arendt, K., Royo, M., Fernández-Monreal, M. et al. PIP3 controls synaptic function by maintaining AMPA receptor clustering at the postsynaptic membrane. Nat Neurosci 13, 36–44 (2010). https://doi.org/10.1038/nn.2462

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