Abstract
This protocol utilizes lipophilic FM dyes to monitor membrane recycling in real time. FM dyes are virtually nonfluorescent in solution but when membrane bound are intensely fluorescent, combined with the flexibility of different emission wavelengths make these dyes an excellent choice for investigating clathrin-mediated endocytosis, among other membrane trafficking and recycling pathways.
The original version of this chapter was revised. A correction to this chapter can be found at https://doi.org/10.1007/978-1-4939-8719-1_19
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Change history
24 October 2018
This book was inadvertently published with the incorrect title as Clathrin-Mediated Endoytosis: Methods and Protocols. This has now been corrected throughout the book to Clathrin-Mediated Endocytosis: Methods and Protocols.
References
Henkel AW, Lubke J, Betz WJ (1996) FM1-43 dye Ultrastructural localization in and release from frog motor nerve terminals. Proc Natl Acad Sci U S A 93:1918–1923
Betz WJ, Mao F, Smith CB (1996) Imaging exocytosis and endocytosis. Curr Opin Neurobiol 6(3):365–371
Ryan TA (2001) Presynaptic imaging techniques. Curr Opin Neurobiol 11(5):544–549
Renger JJ, Egles C, Liu G (2001) A developmental switch in neurotransmitter flux enhances synaptic efficacy by affecting AMPA receptor activation. Neuron 29:469–484
Rea R, Li J, Dharia A et al (2004) Streamlined synaptic vesicle cycle in cone photoreceptor terminals. Neuron 41:755–766
Gordon SJ, Leube RE, Cousin MA (2011) Synaptophysin is required for Synaptobrevin retrieval during synaptic vesicle endocytosis. J Neurosci 31(39):14032–14036
Ryan TA, Smith SJ (1995) Vesicle pool mobilization during action potential firing at hippocampal synapses. Neuron 14(5):983–989
Kim SH, Ryan TA (2009) Synaptic vesicle recycling at CNS synapses without AP-2. J Neurosci 29(12):3865–3874
Ryan TA, Reuter H, Wendland B et al (1993) The kinetics of synaptic vesicle recycling measured at single presynaptic Boutons. Neuron 11(4):713–724
Kay AR, Alfonso A, Alford S et al (1999) Imaging synaptic activity in intact brain and slices with FM1-43 in C. elegans, lamprey and rat. Neuron 24(4):809–817
Clayton EL, Evans GJ, Cousin MA (2008) Bulk synaptic vesicle endocytosis is rapidly triggered during strong stimulation. J Neurosci 28:6627–6632
Clayton EL, Cousin MA (2008) Differential labelling of bulk endocytosis in nerve terminals by FM dyes. Neurochem Int 53:51–55
Acknowledgment
This work was supported by the Medical Research Council (grant number G1002117).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Cousin, M.A., Gordon, S.L., Smillie, K.J. (2018). Using FM Dyes to Monitor Clathrin-Mediated Endocytosis in Primary Neuronal Culture. In: Swan, L. (eds) Clathrin-Mediated Endocytosis. Methods in Molecular Biology, vol 1847. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8719-1_18
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8719-1_18
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8717-7
Online ISBN: 978-1-4939-8719-1
eBook Packages: Springer Protocols