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PLoS One. 2007 Mar 7;2(3):e275.

In vivo bioluminescence imaging of Ca signalling in the brain of Drosophila.

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  • 1Equipe: Bases Neurales des Comportements chez la Drosophile, Laboratoire de Neurobiologie Cellulaire et Moléculaire (NBCM), Centre National de la Recherche Scientifique (CNRS), Unité UPR-9040, Gif-sur-Yvette, France. jean-rene.martin@inaf.cnrs-gif.fr

Abstract

Many different cells' signalling pathways are universally regulated by Ca(2+) concentration [Ca(2+)] rises that have highly variable amplitudes and kinetic properties. Optical imaging can provide the means to characterise both the temporal and spatial aspects of Ca(2+) signals involved in neurophysiological functions. New methods for in vivo imaging of Ca(2+) signalling in the brain of Drosophila are required for probing the different dynamic aspects of this system. In studies here, whole brain Ca(2+) imaging was performed on transgenic flies with targeted expression of the bioluminescent Ca(2+) reporter GFP-aequorin (GA) in different neural structures. A photon counting based technique was used to undertake continuous recordings of cytosolic [Ca(2+)] over hours. Time integrals for reconstructing images and analysis of the data were selected offline according to the signal intensity. This approach allowed a unique Ca(2+) response associated with cholinergic transmission to be identified by whole brain imaging of specific neural structures. Notably, [Ca(2+)] transients in the Mushroom Bodies (MBs) following nicotine stimulation were accompanied by a delayed secondary [Ca(2+)] rise (up to 15 min. later) in the MB lobes. The delayed response was sensitive to thapsigargin, suggesting a role for intra-cellular Ca(2+) stores. Moreover, it was reduced in dunce mutant flies, which are impaired in learning and memory. Bioluminescence imaging is therefore useful for studying Ca(2+) signalling pathways and for functional mapping of neurophysiological processes in the fly brain.

PMID:
17342209
[PubMed - indexed for MEDLINE]
PMCID:
PMC1803028
Free PMC Article
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