Fig. 1

High content monitoring of SV recycling. Hippocampal neurons were transfected with synaptophysin-pHluorin (sypHy) on day in vitro (DIV) 7 and imaged at DIV 13–15. Transfected neurons were challenged with a pulse of impermeant acidic buffer for 1 min (to reveal the surface fraction of sypHy, shaded pink). After returning to imaging buffer for 2 min, neurons were stimulated with a train of 300 action potentials (10 Hz) followed by a 5 min rest period. After this rest period neurons were stimulated with a further train of 400 action potentials (40 Hz). Finally, after a further 3 min neurons were exposed to an alkaline buffer (NH₄Cl, to reveal the total SV pool, blue shaded region). Stimulation is indicated by bars. Mean trace displays the average sypHy fluorescent response of wild-type neurons ± SEM. Traces are ΔF/F₀ and are presented as a fraction of the total SV pool