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. 2022 Jan 8;10:4. doi: 10.1186/s40478-021-01285-5

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© The Author(s) 2022

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 3 — BIN1iso1-induced degeneration is characterized by a strong accumulation of vesicles showing endosomal markers. a Electron microscopy images of retina expressing Luciferase (Ctrl), AmphA, BIN1iso1 and BIN1iso9 from 15-day-old flies. 1 to 2 ommatidia transversal sections are seen on the 4 left images. One photoreceptor neuron is outlined per image, its cytoplasm and nucleus being highlighted in yellow and its rhabdomere in green. Each ommatidium contains 6 outer photoreceptor neurons whose rhabdomeres are organized in a trapezoidal shape, with a 7th rhabdomere in the middle corresponding to one of the two inner photoreceptor neurons. In the BIN1iso1 condition, only 4 rhabdomeres can be seen. In addition, the cytoplasm of the photoreceptor is filled with vesicles, some of them (highlighted in magenta) are very big. These vesicles are surrounded by a single membrane (arrow on the right panel) as compared to the nuclear envelope (double arrow). Their rhabdomere is usually disintegrating (*). b Scheme of the organelles tested in photoreceptor neurons with the markers used. c, d Images of photoreceptor neurons expressing BIN1iso1 and EYFP-tagged endogenous Rab5 and GFP:2xFYVE, as early endosome markers (c), or EYFP-tagged endogenous Rab7 and evi:GFP, as late endosome/multivesicular body markers (d). Rh1 and actin staining, respectively yellow and magenta in the merge image, are used to label retina structure of 15-day-old flies. The photoreceptor neurons expressed Luciferase as a control or BIN1iso1. In the BIN1iso1 conditions, many vesicles were positive for the tested organelle markers (arrows, green staining in merge images). The extracellular inter-rhabdomeric staining for the evi:GFP marker (arrowhead) corresponds to exosomes. e Images of photoreceptor neurons expressing BIN1iso1 stained for BIN1, the photoreceptor neuron plasma membrane Na/K ATPase and actin in 7-day-old flies. BIN1iso1 is localized at the base of the rhabdomeres and around some abnormal vesicles as seen in the inset