In vivo functional calcium imaging of induced or spontaneous activity in the fly brain using a GFP-apoaequorin-based bioluminescent approach

Biochim Biophys Acta. 2013 Jul;1833(7):1632-40. doi: 10.1016/j.bbamcr.2012.12.017. Epub 2012 Dec 31.

Abstract

Different optical imaging techniques have been developed to study neuronal activity with the goal of deciphering the neural code underlying neurophysiological functions. Because of several constraints inherent in these techniques as well as difficulties interpreting the results, the majority of these studies have been dedicated more to sensory modalities than to the spontaneous activity of the central brain. Recently, a novel bioluminescence approach based on GFP-aequorin (GA) (GFP: Green fluorescent Protein), has been developed, allowing us to functionally record in-vivo neuronal activity. Taking advantage of the particular characteristics of GA, which does not require light excitation, we report that we can record induced and/or the spontaneous Ca(2+)-activity continuously over long periods. Targeting GA to the mushrooms-bodies (MBs), a structure implicated in learning/memory and sleep, we have shown that GA is sensitive enough to detect odor-induced Ca(2+)-activity in Kenyon cells (KCs). It has been possible to reveal two particular peaks of spontaneous activity during overnight recording in the MBs. Other peaks of spontaneous activity have been recorded in flies expressing GA pan-neurally. Similarly, expression in the glial cells has revealed that these cells exhibit a cell-autonomous Ca(2+)-activity. These results demonstrate that bioluminescence imaging is a useful tool for studying Ca(2+)-activity in neuronal and/or glial cells and for functional mapping of the neurophysiological processes in the fly brain. These findings provide a framework for investigating the biological meaning of spontaneous neuronal activity. This article is part of a Special Issue entitled: 12th European Symposium on Calcium.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aequorin / genetics
  • Aequorin / metabolism*
  • Animals
  • Animals, Genetically Modified / genetics
  • Apoproteins / genetics
  • Apoproteins / metabolism*
  • Brain / cytology
  • Brain / metabolism*
  • Calcium / metabolism*
  • Calcium Signaling / physiology*
  • Darkness
  • Diagnostic Imaging
  • Drosophila melanogaster / genetics
  • Drosophila melanogaster / growth & development
  • Drosophila melanogaster / metabolism*
  • Female
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism*
  • Light
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Mushroom Bodies / growth & development
  • Mushroom Bodies / metabolism*
  • Neuroglia / cytology
  • Neuroglia / metabolism
  • Neurons / cytology
  • Neurons / metabolism
  • Odorants
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism

Substances

  • Apoproteins
  • Luminescent Proteins
  • Recombinant Proteins
  • apoaequorin
  • Green Fluorescent Proteins
  • Aequorin
  • Calcium