Abstract
The measurement of synaptic vesicle recycling in live neurons transfected with vesicular glutamate transporter fused to pHluorin (vGLUT-pHluorin) allows us to study exocytosis and endocytosis in neurons. When neurons are transfected with this protein we can measure the rate of vesicles fusing and internalizing from the membrane using live total internal reflection fluorescence (TIRF) imaging. Here, we describe transfection, culturing, and imaging of wild-type and αβγ-synuclein knockout hippocampal neurons. This technique can be used to evaluate the effect of different phenotypes and treatments in the physiology of synaptic vesicle recycling in cultured neurons.
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