Optical imaging of calcium transients in neurons and pharyngeal muscle of C. elegans.
Electrophysiology and optical indicators have been used in vertebrate systems to investigate excitable cell firing and calcium transients, but both techniques have been difficult to apply in organisms with powerful reverse genetics. To overcome this limitation, we expressed cameleon proteins, genetically encoded calcium indicators, in the pharyngeal muscle of the nematode worm Caenorhabditis elegans. In intact transgenic animals expressing cameleons, fluorescence ratio changes accompanied muscular contraction, verifying detection of calcium transients. By comparing the magnitude and duration of calcium influx in wild-type and mutant animals, we were able to determine the effects of calcium channel proteins on pharyngeal calcium transients. We also successfully used cameleons to detect electrically evoked calcium transients in individual C. elegans neurons. This technique therefore should have broad applications in analyzing the regulation of excitable cell activity in genetically tractable organisms.
Kerr
R
R
Department of Biology, University of California, San Diego, La Jolla 92093, USA.
Lev-Ram
V
V
Baird
G
G
Vincent
P
P
Tsien
R Y
RY
Schafer
W R
WR
D000818:Animals; D017173:Caenorhabditis elegans / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000235:genetics / Q000502:physiology; D015536:Down-Regulation; D004594:Electrophysiology; D015801:Helminth Proteins / Q000235:genetics; D009124:Muscle Proteins / Q000235:genetics; D009154:Mutation / Q000502:physiology; D009474:Neurons / Q000378:metabolism*; D055095:Optics and Photonics; D010609:Pharyngeal Muscles / Q000294:innervation*; D020033:Protein Isoforms / Q000502:physiology
D029742:Caenorhabditis elegans Proteins; D015220:Calcium Channels; C109164:Egl-19 protein, C elegans; D015801:Helminth Proteins; D009124:Muscle Proteins; D020033:Protein Isoforms; D002118:Calcium