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Monitoring brain activity and behaviour in freely moving Drosophila larvae using bioluminescence

We present a bioluminescence method, based on the calcium-reporter Aequorin (AEQ), that exploits targeted transgenic expression patterns to identify activity of specific neural groups in the larval Drosophila nervous system. We first refine, for intact but constrained larva, the choice of Aequorin t...

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Autores principales: Marescotti, Manuela, Lagogiannis, Konstantinos, Webb, Barbara, Davies, R. Wayne, Armstrong, J. Douglas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6006295/
https://www.ncbi.nlm.nih.gov/pubmed/29915372
http://dx.doi.org/10.1038/s41598-018-27043-7
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author Marescotti, Manuela
Lagogiannis, Konstantinos
Webb, Barbara
Davies, R. Wayne
Armstrong, J. Douglas
author_facet Marescotti, Manuela
Lagogiannis, Konstantinos
Webb, Barbara
Davies, R. Wayne
Armstrong, J. Douglas
author_sort Marescotti, Manuela
collection PubMed
description We present a bioluminescence method, based on the calcium-reporter Aequorin (AEQ), that exploits targeted transgenic expression patterns to identify activity of specific neural groups in the larval Drosophila nervous system. We first refine, for intact but constrained larva, the choice of Aequorin transgene and method of delivery of the co-factor coelenterazine and assay the luminescence signal produced for different neural expression patterns and concentrations of co-factor, using standard photo-counting techniques. We then develop an apparatus that allows simultaneous measurement of this neural signal while video recording the crawling path of an unconstrained animal. The setup also enables delivery and measurement of an olfactory cue (CO(2)) and we demonstrate the ability to record synchronized changes in Kenyon cell activity and crawling speed caused by the stimulus. Our approach is thus shown to be an effective and affordable method for studying the neural basis of behavior in Drosophila larvae.
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spelling pubmed-60062952018-06-26 Monitoring brain activity and behaviour in freely moving Drosophila larvae using bioluminescence Marescotti, Manuela Lagogiannis, Konstantinos Webb, Barbara Davies, R. Wayne Armstrong, J. Douglas Sci Rep Article We present a bioluminescence method, based on the calcium-reporter Aequorin (AEQ), that exploits targeted transgenic expression patterns to identify activity of specific neural groups in the larval Drosophila nervous system. We first refine, for intact but constrained larva, the choice of Aequorin transgene and method of delivery of the co-factor coelenterazine and assay the luminescence signal produced for different neural expression patterns and concentrations of co-factor, using standard photo-counting techniques. We then develop an apparatus that allows simultaneous measurement of this neural signal while video recording the crawling path of an unconstrained animal. The setup also enables delivery and measurement of an olfactory cue (CO(2)) and we demonstrate the ability to record synchronized changes in Kenyon cell activity and crawling speed caused by the stimulus. Our approach is thus shown to be an effective and affordable method for studying the neural basis of behavior in Drosophila larvae. Nature Publishing Group UK 2018-06-18 /pmc/articles/PMC6006295/ /pubmed/29915372 http://dx.doi.org/10.1038/s41598-018-27043-7 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Marescotti, Manuela
Lagogiannis, Konstantinos
Webb, Barbara
Davies, R. Wayne
Armstrong, J. Douglas
Monitoring brain activity and behaviour in freely moving Drosophila larvae using bioluminescence
title Monitoring brain activity and behaviour in freely moving Drosophila larvae using bioluminescence
title_full Monitoring brain activity and behaviour in freely moving Drosophila larvae using bioluminescence
title_fullStr Monitoring brain activity and behaviour in freely moving Drosophila larvae using bioluminescence
title_full_unstemmed Monitoring brain activity and behaviour in freely moving Drosophila larvae using bioluminescence
title_short Monitoring brain activity and behaviour in freely moving Drosophila larvae using bioluminescence
title_sort monitoring brain activity and behaviour in freely moving drosophila larvae using bioluminescence
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6006295/
https://www.ncbi.nlm.nih.gov/pubmed/29915372
http://dx.doi.org/10.1038/s41598-018-27043-7
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