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Fig. 5. Ultrastructure analysis of MB calyx. (A) Ultrastructure analysis of larval (A2,A3) and 6 hours APF calyx (A4,A5) using GH146 driven EM marker CD2::HRP labeling PPN axons. (A1) Illustration of electron micrograph (EM) section plane using confocal Z projection of larval GH146-GAL4 driving mCD8::GFP, which labels the AL, inner antennocerebral tract and calyx with a dashed box showing the approximate location of EM section through the calyx. (A2) 4000x EM of the larval calyx showing the punctate labeling pattern with labeled boutons toward the outside of the calyx. Asterisks denote large presynaptic boutons. (A3) 40,000x EM of labeled larval PPN presynaptic terminals (filled arrowheads) apposing small unlabeled dendritic profiles postsynaptically. Open arrowheads point to the profile of the entire presynaptic bouton. (A4) 4000x EM of 6 hours APF degenerating PPN presynaptic boutons in the calyx. (A5) 40,000x EM at 6 hours APF with open arrowheads pointing to the border of a labeled presynaptic bouton and surrounding glia cytoplasm. (B) Ultrastructure analysis of larval (B2,B3) and 6 hours APF calyx (B4,B5) using GAL4-201Y driving CD2::HRP labeling MB {gamma} neuron dendrites. (B1) Illustration of EM section plane using confocal Z projection of larval 201Y-GAL4 driving mCD8::GFP, which labels dendrites in the calyx, the axon peduncle, and medial and dorsal axon lobes. Dashed box shows approximate location of EM section through the calyx. (B2) 4000x EM of the larval calyx showing the concentric labeling pattern, with labeled dendrites toward the outside of the calyx. Asterisks denote large presynaptic boutons. (B3) 40,000x EM of labeled larval dendritic calyx showing unlabeled PPN presynaptic terminals (filled arrowheads) apposing small labeled dendritic profiles postsynaptically. (B4) 4000x EM of 6 hours APF MB {gamma} neuron labeled calyx showing degeneration of larval concentric labeling. (B5) 40,000x EM at 6 hours APF with an arrow specifying a degenerating synapse full of synaptic vesicles surrounded by labeled dendritic profiles, which appear to be engulfed by a glia (open arrowheads pointing to the border of engulfed synaptic profiles and glia cytoplasm). Dotted lines in A2,A4,B2,B4 denote the border of the calyx. Scale bars: 50 µm in A1,B1; 10 µm in A2,A4,B2,B4; 1 µm in A3,A5,B3,B5. c, calyx; d, dorsal axon lobe; iACT, inner antennocerebral tract; m, medial axon lobe; p, axon peduncle.





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