title: In vivo neuronal calcium imaging in C. elegans.
abstract: The nematode worm C. elegans is an ideal model organism for relatively simple, low cost neuronal imaging in vivo. Its small transparent body and simple, well-characterized nervous system allows identification and fluorescence imaging of any neuron within the intact animal. Simple immobilization techniques with minimal impact on the animal's physiology allow extended time-lapse imaging. The development of genetically-encoded calcium sensitive fluorophores such as cameleon and GCaMP allow in vivo imaging of neuronal calcium relating both cell physiology and neuronal activity. Numerous transgenic strains expressing these fluorophores in specific neurons are readily available or can be constructed using well-established techniques. Here, we describe detailed procedures for measuring calcium dynamics within a single neuron in vivo using both GCaMP and cameleon. We discuss advantages and disadvantages of both as well as various methods of sample preparation (animal immobilization) and image analysis. Finally, we present results from two experiments: 1) Using GCaMP to measure the sensory response of a specific neuron to an external electrical field and 2) Using cameleon to measure the physiological calcium response of a neuron to traumatic laser damage. Calcium imaging techniques such as these are used extensively in C. elegans and have been extended to measurements in freely moving animals, multiple neurons simultaneously and comparison across genetic backgrounds. C. elegans presents a robust and flexible system for in vivo neuronal imaging with advantages over other model systems in technical simplicity and cost.
authors.affiliation: Department of Physiology and Biophysics, Boston University School of Medicine, USA.
authors.forename: Samuel H
authors.initials: SH
authors.lastname: Chung
authors.forename: Lin
authors.initials: L
authors.lastname: Sun
authors.forename: Christopher V
authors.initials: CV
authors.lastname: Gabel
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D017173:Caenorhabditis elegans; D002118:Calcium / Q000032:analysis / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000737:chemistry; D008856:Microscopy, Fluorescence / Q000379:methods; D009474:Neurons / Q000737:chemistry / Q000378:metabolism*; D061848:Optical Imaging / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D012149:Restraint, Physical
chemical_list: D005456:Fluorescent Dyes; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D002118:Calcium