Two-photon calcium imaging from head-fixed Drosophila during optomotor walking behavior.
Drosophila melanogaster is a model organism rich in genetic tools to manipulate and identify neural circuits involved in specific behaviors. Here we present a technique for two-photon calcium imaging in the central brain of head-fixed Drosophila walking on an air-supported ball. The ball's motion is tracked at high resolution and can be treated as a proxy for the fly's own movements. We used the genetically encoded calcium sensor, GCaMP3.0, to record from important elements of the motion-processing pathway, the horizontal-system lobula plate tangential cells (LPTCs) in the fly optic lobe. We presented motion stimuli to the tethered fly and found that calcium transients in horizontal-system neurons correlated with robust optomotor behavior during walking. Our technique allows both behavior and physiology in identified neurons to be monitored in a genetic model organism with an extensive repertoire of walking behaviors.
Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA.
Johannes D
JD
Seelig
M Eugenia
ME
Chiappe
Gus K
GK
Lott
Anirban
A
Dutta
Jason E
JE
Osborne
Michael B
MB
Reiser
Vivek
V
Jayaraman
D000818:Animals; D001921:Brain / Q000166:cytology / Q000502:physiology; D002118:Calcium / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000502:physiology*; D005453:Fluorescence; D049452:Green Fluorescent Proteins; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation* / Q000379:methods*; D009038:Motion; D009043:Motor Activity / Q000502:physiology*; D009474:Neurons / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology; D016138:Walking / Q000502:physiology*
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D002118:Calcium