Long-term depression in neurons involves temporal and ultra-structural dynamics of phosphatidylinositol-4,5-bisphosphate relying on PIP5K, PTEN and PLC

Synaptic plasticity involves proper establishment and rearrangement of structural and functional microdomains. Yet, visualization of the underlying lipid cues proved challenging. Applying a combination of rapid cryofixation, membrane freeze-fracturing, immunogold labeling and electron microscopy, we visualize and quantitatively determine the changes and the distribution of phosphatidylinositol-4,5-bisphosphate (PIP2) in the plasma membrane of dendritic spines and subareas thereof at ultra-high resolution. These efforts unravel distinct phases of PIP2 signals during induction of long-term depression (LTD). During the first minutes PIP2 rapidly increases in a PIP5K-dependent manner forming nanoclusters. PTEN contributes to a second phase of PIP2 accumulation. The transiently increased PIP2 signals are restricted to upper and middle spine heads. Finally, PLC-dependent PIP2 degradation provides timely termination of PIP2 cues during LTD induction. Together, this work unravels the spatial and temporal cues set by PIP2 during different phases after LTD induction and dissects the molecular mechanisms underlying the observed PIP2 dynamics. Ultrahigh-resolution views of PIP2 in freeze-fractured plasma membranes of dendritic spines during induction of long-term depression reveal spatial and temporal cues set by PIP2 and reveal the molecular mechanisms behind the observed PIP2 dynamics

Automated summary

Using advanced imaging techniques, this study visualizes and quantitatively determines the changes and distribution of PIP2 in dendritic spines during LTD induction. It reveals distinct phases of PIP2 signals and dissects the molecular mechanisms underlying the observed PIP2 dynamics.