A thermogenetics protocol for detecting gap junction channels in Drosophila egg chambers.
A thermogenetics protocol for detecting gap junction channels in Drosophila egg chambers.
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DOI：
10.1016/j.xpro.2020.100269
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发表时间：
2021-03-19
期刊：
影响因子：
--
通讯作者：
Montell D
中科院分区：
文献类型：
--
作者：
Miao G;Montell D
在本实验中，我们采用“热遗传学”的方法来检测果蝇卵室中细胞间的功能性缝隙连接通道。我们使用Gal 4在种系中表达热敏阳离子通道TrpA 1-A，并且使用莱克萨/LexAop系统在所有卵泡细胞中表达荧光[Ca 2 +]传感器（GCaMP 6）。如果间隙连接连接生殖细胞和卵泡细胞，我们预计温度依赖性TRPA 1-A依赖性阳离子从培养基流入生殖细胞，导致卵泡细胞中的GCaMP信号。有关本方案使用和执行的完整详细信息，请参阅。可以使用“热遗传学”方法检测功能性间隙连接通道。使用Gal 4在种系中表达热敏阳离子通道TrpA 1-A。使用莱克萨在卵泡细胞中表达荧光[Ca 2 +]传感器（GCaMP 6 s）。温度升高时可以检测TRPA 1-A依赖性阳离子流入。在该方案中，我们采用“热遗传学”方法来检测果蝇卵室中细胞之间的功能性间隙连接通道。我们使用Gal 4在种系中表达热敏阳离子通道TrpA 1-A，并且使用莱克萨/LexAop系统在所有卵泡细胞中表达荧光[Ca 2 +]传感器（GCaMP 6）。如果间隙连接连接生殖细胞和卵泡细胞，我们预计温度依赖性TRPA 1-A依赖性阳离子从培养基流入生殖细胞，导致卵泡细胞中的GCaMP信号。
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 [Ca2+] 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. Functional gap junction channels can be detected using a “thermogenetics” approach The thermosensitive cation channel TrpA1-A is expressed in the germline using Gal4 The fluorescent [Ca2+] sensor (GCaMP6s) is expressed in follicle cells using LexA A TRPA1-A dependent cation influx can be detected upon temperature increase 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 [Ca2+] 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.
影响因子：
11.8
作者：
Miao, Guangxia;Godt, Dorothea;Montell, Denise J.
通讯作者：
Montell, Denise J.
影响因子：
14.8
作者：
Prasad, Mohit;Jang, Anna C-C;Montell, Denise J.
通讯作者：
Montell, Denise J.