Downregulation of Plasma Membrane Ca2+ ATPase driven by tyrosine hydroxylase-Gal4 reduces Drosophila lifespan independently of effects in neurons

In Drosophila melanogaster, several Gal4 drivers are used to direct gene/RNAi expression to different dopaminergic neuronal clusters. We previously developed a fly model of Parkinson's disease, in which dopaminergic neurons had elevated cytosolic Ca2+ due to the expression of a Plasma Membrane Ca2+ ATPase (PMCA) RNAi under the thyroxine hydroxylase (TH)-Gal4 driver. Surprisingly, TH-Gal4>PMCA(RNAi) flies died earlier compared to controls and showed swelling in the abdominal area. Flies expressing the PMCA(RNAi) under other TH drivers also showed such swelling and shorter lifespan. Considering that TH-Gal4 is also expressed in the gut, we proposed to suppress the expression specifically in the nervous system, while maintaining the activation in the gut. Therefore, we expressed Gal80 under the direction of the panneuronal synaptobrevin (nSyb) promoter in the context of TH-Gal4. nSyb-Gal80; TH-Gal4>PMCA(RNAi) flies showed the same reduction of survival as TH-Gal4>PMCA(RNAi) flies, meaning that the phenotype of abdomen swelling and reduced survival could be due to the expression of the PMCA(RNAi) in the gut. In perimortem stages TH-Gal4>PMCA(RNAi) guts had alteration in the proventriculi and crops. The proventriculi appeared to lose cells and collapse on itself, and the crop increased its size several times with the appearance of cellular accumulations at its entrance. No altered expression or phenotype was observed in flies expressing PMCA(RNAi) in the dopaminergic PAM cluster (PAM-Gal4>PMCA(RNAi)). In this work we show the importance of checking the global expression of each promoter and the relevance of the inhibition of PMCA expression in the gut.

Automated summary

In fruit flies, Gal4 drivers are used to control gene/RNAi expression in different dopaminergic neuronal clusters. A Parkinson's disease fly model was developed with elevated cytosolic Ca2+ in dopaminergic neurons due to the expression of PMCA(RNAi) under TH-Gal4 driver. Surprisingly, the flies showed abdominal swelling and shorter lifespan, suggesting a potential role of gut expression. Suppressing PMCA(RNAi) expression specifically in the nervous system did not improve the phenotype, indicating that the gut expression might be responsible. Evaluation: 8/10.