Imaging synaptic activity in intact brain and slices with FM1-43 in C. elegans, lamprey, and rat

Alan R. Kay, Aixa Alfonso, Simon Alford, Hollis T. Cline, Andrea M. Holgado, Bert Sakmann, Vladislav Snitsarev, Thomas P. Stricker, Michiko Takahashi, Ling Gang Wu

Research output: Contribution to journalArticle

120 Citations (Scopus)

Abstract

The fluorescent probe FM1-43 has been used extensively for imaging vesicle recycling; however, high nonspecific adsorption resulting in elevated background levels has precluded its use in certain tissues, notably brain slices. We have found that a sulfobutylated derivative of β-cyclodextrin (ADVASEP-7) has a higher affinity for FM1-43 than the plasma membrane. ADVASEP-7 was used as a carrier to remove FM1-43 nonspecifically bound to the outer leaflet of the plasma membrane or extracellular molecules, significantly reducing background staining. This has enabled us to visualize synaptic vesicle recycling in the nematode C. elegans, intact lamprey spinal cord, and rat brain slices.

Original languageEnglish
Pages (from-to)809-817
Number of pages9
JournalNeuron
Volume24
Issue number4
DOIs
StatePublished - 1 Jan 1999

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Lampreys
Brain
Cell Membrane
Synaptic Vesicles
Cyclodextrins
Recycling
Fluorescent Dyes
Adsorption
Spinal Cord
Staining and Labeling
FM1 43
ADVASEP 7

Cite this

Kay, A. R., Alfonso, A., Alford, S., Cline, H. T., Holgado, A. M., Sakmann, B., ... Wu, L. G. (1999). Imaging synaptic activity in intact brain and slices with FM1-43 in C. elegans, lamprey, and rat. Neuron, 24(4), 809-817. https://doi.org/10.1016/S0896-6273(00)81029-6
Kay, Alan R. ; Alfonso, Aixa ; Alford, Simon ; Cline, Hollis T. ; Holgado, Andrea M. ; Sakmann, Bert ; Snitsarev, Vladislav ; Stricker, Thomas P. ; Takahashi, Michiko ; Wu, Ling Gang. / Imaging synaptic activity in intact brain and slices with FM1-43 in C. elegans, lamprey, and rat. In: Neuron. 1999 ; Vol. 24, No. 4. pp. 809-817.
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Kay, AR, Alfonso, A, Alford, S, Cline, HT, Holgado, AM, Sakmann, B, Snitsarev, V, Stricker, TP, Takahashi, M & Wu, LG 1999, 'Imaging synaptic activity in intact brain and slices with FM1-43 in C. elegans, lamprey, and rat', Neuron, vol. 24, no. 4, pp. 809-817. https://doi.org/10.1016/S0896-6273(00)81029-6

Imaging synaptic activity in intact brain and slices with FM1-43 in C. elegans, lamprey, and rat. / Kay, Alan R.; Alfonso, Aixa; Alford, Simon; Cline, Hollis T.; Holgado, Andrea M.; Sakmann, Bert; Snitsarev, Vladislav; Stricker, Thomas P.; Takahashi, Michiko; Wu, Ling Gang.

In: Neuron, Vol. 24, No. 4, 01.01.1999, p. 809-817.

Research output: Contribution to journalArticle

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Kay AR, Alfonso A, Alford S, Cline HT, Holgado AM, Sakmann B et al. Imaging synaptic activity in intact brain and slices with FM1-43 in C. elegans, lamprey, and rat. Neuron. 1999 Jan 1;24(4):809-817. https://doi.org/10.1016/S0896-6273(00)81029-6