Validation of marker genes for lamina-specific glial subtypes of the young adult optic lobe.

(A, B)Chiasm(R53H12)>GFP adult optic lobe, showing expression in chiasm and distal satellite glia. Obp18a expression (magenta) was detected by in situ HCR in most GFP positive (green) nuclei of chiasm and distal satellite glia. (C, D) Perineurial(R85G01)>GFP adult optic lobe where JhI-21 (Juvenile hormone Inducible-21) expression (magenta) was detected by in situ HCR, specifically in chalice glia. (E, F) Perineurial(R85G01)>CD8::GFP adult optic lobe where CG13003 expression (magenta) was detected by in situ HCR in fenestrated glia. (G) Chiasm(R53H12)>GFP adult optic lobe where CG7135 expression (magenta) was detected by in situ HCR in chiasm, chalice, and proximal satellite glia. (H) Fas3 (magenta) antibody staining of Fenestrated(R47G01)>CD8::GFP adult optic lobe, showed expression in the lamina surface glia layer below the fenestrated glia, pseudo-cartridge glia, in the chalice, and proximal satellite glia. (I) Perineurial(R85G01)>CD8::GFP adult optic lobe where Cyp311a1 (Cytochrome P450 311a1) expression (magenta) was detected by in situ HCR, specifically in chalice glia. (J) Fenestrated(R47G01)>nls::GFP adult optic lobe where GstE9 (Glutathione S transferase E9) expression (magenta) was detected by in situ HCR in fenestrated and pseudo-cartridge glia. (K) Proximal satellite(R46H12)>nls::GFP adult optic lobe where Ndae1 (Na+-driven anion exchanger 1) expression (magenta) was detected by in situ HCR in proximal satellite glia, as well as in L1, L2, L3, and L4 lamina neurons, as predicted by the neuronal scRNA-seq data from [57]. (L) Large MCFO clones labelled by ltl-Gal4, marked distal satellite (bar) and fenestrated glia (arrow). HRP in white. (M) Chiasm(R53H12)>CD8::GFP adult optic lobe where Tsf1 expression (magenta) was detected by in situ HCR in proximal and distal satellite and chiasm glia. (N) Proximal satellite(R46H12)>CD8::GFP adult optic lobe where GILT1 expression (magenta) was detected by in situ HCR in proximal satellite glia, as well as in T1 medulla neurons, as predicted by the neuronal scRNA-seq data from [57]. All panels are single focal planes. Dashed lines outline the neuropils and scale bars are 20 μm. (TIF)