Neuropeptide T2A-GAL4 knock-ins illustrate practical considerations for interpreting GAL4 reporter patterns in Drosophila
Jinnai, K.; Ozawa, K.; Yajima, K.; Tanimoto, H.; Kondo, S.
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Transgenic GAL4 lines are widely used in Drosophila to label and manipulate specific cell types, but the resulting reporter expression patterns do not always represent ongoing driver activity. We generated a collection of neuropeptide T2A-GAL4 knock-ins and characterized their expression by native-fluorescence imaging using UAS-CD8::GFP and a Brp-miRFP680 knock-in, eliminating the need for immunostaining. During characterization of these lines, we observed cases in which adult GFP patterns did not match the on-going GAL4 activity, as well as cases in which GAL4 produced adverse cellular effects. Using temperature-sensitive GAL80, we found that adult GFP expression in certain neurons driven by CCHa2-T2A-GAL4 and Dsk-T2A-GAL4 depended on GAL4 activity before eclosion and was not reproduced by adult-only induction. Several GAL4 lines also showed dominant developmental phenotypes. Heterozygous Burs-T2A-GAL4 animals, for example, displayed wing-expansion defects that were fully suppressed by GAL80. Severe wing defects were not accompanied by a reduction in the number of reporter-labeled neurons before eclosion, but the morphology of these neurons were affected. These observations illustrate how developmental driver history and GAL4-associated cellular effects can influence reporter patterns in the adult nervous system.
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