Prominent involvement of acetylcholine dynamics in stable olfactory representation across the Drosophila brain
- Jiaqi Fan
- Yuling Wang
- Lingbo Li
- Jing He
- Zhifeng Zhao
- Fei Deng
- Guochuan Li
- Xinyang Li
- Yiliang Zhou
- Jiayin Zhao
- Ning Huang
- Yixin Hu
- Yulong Li
- Jiamin Wu
- Lu Fang
- Qionghai Dai
2025-09-30
Despite the vital role of neuromodulators and neurotransmitters in the neural system, their spatiotemporal correlation with neuronal activities across multiple brain regions remain unclear. Here, we employed two-photon synthetic aperture microscopy (2pSAM) and neurochemical indicators to simultaneously record calcium and acetylcholine (ACh)/5-HT dynamics across multiple regions of the Drosophila brain over 2 h. Presenting 3 different odors across multiple trials, our analyses revealed signal-specific differences in responsiveness, functional connectivity, and odor classification accuracy across the brain. We further constructed low-dimensional manifolds to characterize the global odor-related dynamics. Incorporating both calcium and ACh signals enhanced odor classification accuracy in the global low-dimensional manifold and in specific brain regions where their functional connectivity network features exhibited complementary patterns. Moreover, ACh dynamics demonstrated relatively stable temporal characteristics compared to calcium and 5-HT. These results suggest the potential contribution of ACh to consistent odor representations and illustrate the utility of multi-signal imaging in studying neural computation.