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

Kay AR, Alfonso A, Alford S, Cline HT, Holgado AM, Sakmann B, Snitsarev VA, Stricker TP, Takahashi M, Wu LG
Neuron. 1999 24 (4): 809-17

PMID: 10624945

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 beta-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.

MeSH Terms (20)

Animals Axons Brain Brain Stem Caenorhabditis elegans Cyclodextrins Drug Carriers Fluorescent Dyes Image Processing, Computer-Assisted In Vitro Techniques Lampreys Liposomes Male Pyridinium Compounds Quaternary Ammonium Compounds Rats Rats, Long-Evans Rats, Wistar Spinal Cord Synapses

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