The driver CHA-Gal4 is used to simultaneously express GFP and a nuclear RFP in cholinergic neurons. In cells with short processes, both GFP and RFP expression are more tightly localized in the nucleus, resulting in cells with yellow nuclei. In contrast, in cells with long processes, GFP spreads out resulting in nuclei with relatively stronger red fluorescence. On top of this, nuclear localization of DATI transcription factor is labeled in blue. This color-code allows the detection of four distinct classes of neurons: DATI- CHA+ neurons appear in green; DATI+ CHA- neurons in blue; DATI+ CHA+ neurons with short processes in light blue, and DATI+CHA+ neurons with long processes in magenta. Processes of CHA+ neurons appear in green. See Schinaman et al.
Image Credit: Joseph Schinaman and Rui Sousa-Neves
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