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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 |
Sumario: | 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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