Assessment of the contribution of the plasma membrane calcium ATPase, PMCA, calcium transporter to synapse function using patch clamp electrophysiology and fast calcium imaging.

The plasma membrane calcium ATPase, or PMCA, functions to extrude calcium out of cells as a key component necessary for adequate calcium homeostasis in all cells. However, calcium is particularly important at synapses between neurons, where communication relies on the controlled rise and fall in presynaptic calcium that precedes the release of neurotransmitter. Here we show how to infer the real-time contribution of PMCA-mediated calcium extrusion to this presynaptic calcium dynamic and how this influences the properties of the synapse. To do this we have taken advantage of a well-studied synapse in the cerebellum. We use electrophysiology to assess the timing of short-term facilitation at this synapse in the presence and absence of PMCA2 using PMCA2 knockout mice and pharmacology and fast calcium imaging to measure the presynaptic calcium dynamics. These approaches are all highly applicable to other synapses and can help determine the contribution of PMCA, and other transporters or exchangers, to the calcium dynamics that underpin reliable synaptic transmission.