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A thermogenetics protocol for detecting gap junction channels in Drosophila egg chambers

In this protocol, we took a “thermogenetics” approach to detect functional gap junction channels between cells in Drosophila egg chambers. We expressed the thermosensitive cation channel TrpA1-A in the germline using Gal4, and a fluorescent [Ca(2+)] sensor (GCaMP6), in all follicle cells using the L...

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Detalles Bibliográficos
Autores principales: Miao, Guangxia, Montell, Denise
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7811055/
https://www.ncbi.nlm.nih.gov/pubmed/33490985
http://dx.doi.org/10.1016/j.xpro.2020.100269
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author Miao, Guangxia
Montell, Denise
author_facet Miao, Guangxia
Montell, Denise
author_sort Miao, Guangxia
collection PubMed
description In this protocol, we took a “thermogenetics” approach to detect functional gap junction channels between cells in Drosophila egg chambers. We expressed the thermosensitive cation channel TrpA1-A in the germline using Gal4, and a fluorescent [Ca(2+)] sensor (GCaMP6), in all follicle cells using the LexA/LexAop system. If gap junctions connect germ cells and follicle cells, we expect a temperature-dependent TRPA1-A-dependent cation influx into the germline from the culture medium to result in a GCaMP signal in follicle cells. For complete details on the use and execution of this protocol, please refer to Miao et al. (2020).
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spelling pubmed-78110552021-01-22 A thermogenetics protocol for detecting gap junction channels in Drosophila egg chambers Miao, Guangxia Montell, Denise STAR Protoc Protocol In this protocol, we took a “thermogenetics” approach to detect functional gap junction channels between cells in Drosophila egg chambers. We expressed the thermosensitive cation channel TrpA1-A in the germline using Gal4, and a fluorescent [Ca(2+)] sensor (GCaMP6), in all follicle cells using the LexA/LexAop system. If gap junctions connect germ cells and follicle cells, we expect a temperature-dependent TRPA1-A-dependent cation influx into the germline from the culture medium to result in a GCaMP signal in follicle cells. For complete details on the use and execution of this protocol, please refer to Miao et al. (2020). Elsevier 2021-01-14 /pmc/articles/PMC7811055/ /pubmed/33490985 http://dx.doi.org/10.1016/j.xpro.2020.100269 Text en © 2021 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Miao, Guangxia
Montell, Denise
A thermogenetics protocol for detecting gap junction channels in Drosophila egg chambers
title A thermogenetics protocol for detecting gap junction channels in Drosophila egg chambers
title_full A thermogenetics protocol for detecting gap junction channels in Drosophila egg chambers
title_fullStr A thermogenetics protocol for detecting gap junction channels in Drosophila egg chambers
title_full_unstemmed A thermogenetics protocol for detecting gap junction channels in Drosophila egg chambers
title_short A thermogenetics protocol for detecting gap junction channels in Drosophila egg chambers
title_sort thermogenetics protocol for detecting gap junction channels in drosophila egg chambers
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7811055/
https://www.ncbi.nlm.nih.gov/pubmed/33490985
http://dx.doi.org/10.1016/j.xpro.2020.100269
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