D, Isolated Munc18-1cKO and WT (control) neurons infected at DIV8 with Cre-EGFP were tested for viability (+ = viable, – = non-viable) and for SV exocytosis using Synaptophysin-pHluorin (SypHy). The typical trace shows the change in fluorescence (ΔF) over time measured from the indicated exocytosis event. C, Disappearance of a NPY-mCherry punctum visualized by two still frames: one before and one after the indicated exocytosis event (red arrowhead, same event as in B). Below, Schematic model of an NPY-mCherry release event. During high-frequency train-stimulation indicated by blue bars (16 trains of 50 APs at 50 Hz), NPY-mCherry release events are visible as an abrupt termination of the line, marked by arrowheads. B, The kymograph shows NPY-mCherry fluorescence in an axonal stretch over time. A, Schematic representation of a primary mouse hippocampal neuron grown on a glia microisland. Munc18-1 is essential for DCV exocytosis in neurons.
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