Imaging synaptic activity in intact brain and slices with FM1-43 in C. elegans, lamprey, and rat.
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.
Kay
A R
AR
Department of Biological Sciences, University of Iowa, Iowa City 52242, USA. alan-kay@uiowa.edu
Alfonso
A
A
Alford
S
S
Cline
H T
HT
Holgado
A M
AM
Sakmann
B
B
Snitsarev
V A
VA
Stricker
T P
TP
Takahashi
M
M
Wu
L G
LG
D000818:Animals; D001369:Axons / Q000502:physiology; D001921:Brain / Q000033:anatomy & histology / Q000166:cytology / Q000502:physiology*; D001933:Brain Stem / Q000033:anatomy & histology / Q000502:physiology; D017173:Caenorhabditis elegans / Q000502:physiology*; D003505:Cyclodextrins / Q000494:pharmacology; D004337:Drug Carriers; D005456:Fluorescent Dyes; D007091:Image Processing, Computer-Assisted; D066298:In Vitro Techniques; D007798:Lampreys / Q000502:physiology*; D008081:Liposomes; D008297:Male; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D051381:Rats; D020318:Rats, Long-Evans; D017208:Rats, Wistar; D013116:Spinal Cord / Q000033:anatomy & histology / Q000502:physiology; D013569:Synapses / Q000502:physiology* / Q000648:ultrastructure
C403046:ADVASEP 7; D003505:Cyclodextrins; D004337:Drug Carriers; C073804:FM1 43; D005456:Fluorescent Dyes; D008081:Liposomes; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds