Identification of neural circuits by imaging coherent electrical activity with FRET-based dyes.
We show that neurons that underlie rhythmic patterns of electrical output may be identified by optical imaging and frequency-domain analysis. Our contrast agent is a two-component dye system in which changes in membrane potential modulate the relative emission between a pair of fluorophores. We demonstrate our methods with the circuit responsible for fictive swimming in the isolated leech nerve cord. The output of a motor neuron provides a reference signal for the phase-sensitive detection of changes in fluorescence from individual neurons in a ganglion. We identify known and possibly novel neurons that participate in the swim rhythm and determine their phases within a cycle. A variant of this approach is used to identify the postsynaptic followers of intracellularly stimulated neurons.
Cacciatore
T W
TW
Group in Neurosciences, University of California, San Diego, La Jolla 92093, USA.
Brodfuehrer
P D
PD
Gonzalez
J E
JE
Jiang
T
T
Adams
S R
SR
Tsien
R Y
RY
Kristan
W B
WB
Kleinfeld
D
D
D000818:Animals; D002462:Cell Membrane / Q000502:physiology; D004594:Electrophysiology / Q000379:methods*; D005456:Fluorescent Dyes; D017952:Ganglia, Invertebrate / Q000166:cytology / Q000502:physiology; D007395:Interneurons / Q000166:cytology / Q000502:physiology; D007865:Leeches; D008856:Microscopy, Fluorescence / Q000379:methods; D009046:Motor Neurons / Q000166:cytology* / Q000502:physiology*; D009068:Movement / Q000502:physiology; D009420:Nervous System / Q000166:cytology; D009434:Neural Pathways; D013550:Swimming / Q000502:physiology
D005456:Fluorescent Dyes