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

Abstract

Summary: 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 Miao et al. (2020).

Keywords

• Cell biology
• Genetics
• Microscopy
• Model organisms