Fly-eFISH and PTR are using EASI-FISH to map NPs throughout the CNS in male and female flies to produce an atlas at the resolution of brain areas. This effort will help validate transcriptomic data and the EASI-FISH technique. It will tell us which NPs are candidates to be expressed in individual split-GAL4 lines.
The Project Pipeline Support (PPS) team is further scaling up fly NP EASI-FISH for Fly-eFISH and applying it to a wider range of CNS cell types. The project will focus on cell types targeted by specific split-GAL4 lines. Because of the large number of cell types, the examined set will be further restricted:
- Cell types with published transcriptomic data will provide a basis for comparison and validation in some cases.
- Fly-eFISH will collaborate with Janelia labs and visitors to leverage anatomical expertise in regions of interest for evaluating EASI-FISH images.
- Neuropeptides are typically released by neurons with dense core vesicles (DCVs) visible in EM. Connectomics Annotation will assist in evaluating candidate cell types for the presence of DCVs.
PTR, PPS, and the Tillberg lab are collaborating on further improvements to EASI-FISH signal quality. Future work in collaboration with the Rubin lab has been proposed for examining NP receptors and spatial aspects of NP and gap junction signaling in the CNS, as well as NP signaling between the fly CNS and gut.