Abstract
This protocol comprises the entire process of fluorescent measurement of vesicle recycling using the probe SynaptopHluorin, a pH-dependent GFP variant whose fluorescence increases at the synapse upon vesicle release due to fluorescence quenching in acidic vesicles. This technique provides a genetic tool to monitor synaptic vesicle recycling in real time in cultured hippocampal neurons.
Materials and Reagents
Equipment
Software
Procedure
Recipes
Acknowledgments
The methods were adapted from (Li et al., 2013) Techniques were also adapted from all of the references cited. The authors wish to thank Miesenbock, G. (Miesenbock et al., 1998) for providing the synaptopHluorin construct for these studies. This work was supported by grant NIH NS064967 to EAJ.
References
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