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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...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
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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). |
format | Online Article Text |
id | pubmed-7811055 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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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