Synapses are extraordinary signaling machines. Thousands of proteins precisely and uniquely arranged within the pre- and postsynaptic compartments of each synapse govern neuronal communication and mediate plasticity in the nervous system. One of the principle goals in neuroscience has long been to measure synaptic structure and protein organization, and to monitor mechanisms of change at individual synapses in living cells. Since synapses are very small, highly dynamic, and densely packed within light-scattering medium this goal has been difficult to attain. Recent advances in microscopic techniques are likely to change that by improving spatial resolution by an order of magnitude, thus making it possible to investigate synaptic physiology and biochemistry at individual synapses even at the level of single molecules. We will review our progress in using superresolution STED microscopy to study the dynamics of synaptic structures using lifeact to label filamentous actin with subsynaptic resolution in live-cell imaging experiments in brain slices. |