Fig. 3: Cell type complement of the E. berryi adult optic lobe.

a, UMAP of 26,436 cells from an adult E. berryi optic lobe highlighting the location of each cell population. b, Violin plots indicating genes specifically expressed in sets of key cell clusters. c, Antibody staining demonstrating the location of FMRF-amide peptide, acetylated tubulin and 4′,6-diamidino-2-phenylindole (DAPI) in the E. berryi retina. d, Toluidine blue staining of a cross-section of the optic lobe. Inset: an overview of the optic lobe. e–l′, Fluorescence imaging of HCR stainings: pan-neuronal marker Sy65 (synaptotagmin) expression demonstrating the location of neuronal cells in both the optic lobe cortex and the medulla (e); dopamine synthesis enzyme Ty3h (tyrosine hydroxylase) expression in a subset of neuronal cells of both the optic lobe cortex and the medulla (f); cholinergic cell marker, acetylcholine esterase, Aces-2, expression in cells of the optic lobe medulla (g); kainate glutamate receptor Grik2-2 expression in the optic lobe outer granule layer (ogl) and in large cells of the medulla (h); kainate glutamate receptor Grik2-4 expression in the optic lobe outer granule layer and punctate expression in the medulla (i); Six4 expression in the optic lobe outer granule layer of the cortex (j); FMRF ligand transcript localization using HCR in whole mounts (showing an overview of FMRF ligand producing cells arranged in a rosette formation located in deep cells of the medulla (k) and a magnification of FMRF-expressing cell clusters in the medulla (k′)); the expression of glial marker Eaa1-2 on cryosection (l) and in whole-mount staining (l′), demonstrating Eaa1-2 expression in large cells of the medulla including large cells located in the palisade layer of the medulla (located directly beneath the inner granule layer of cortex) and a punctate pattern in cells that may be adjacent to projections of cells in the inner plexiform layer and of axonal projections in the medulla. Stainings were reproduced at least three times. igl, inner granule layer; ipl, inner plexiform layer; md, medulla; ogl, outer granule layer. Scale bars, 100 µm (c) and 50 µm (d–l′).