2026-10-11T04:11:05Z https://api.figshare.com/v2/oai

oai:figshare.com:article/34017682 2026-09-28T17:52:51Z category_12 category_13 category_15 category_272 category_734 category_61 category_64 category_128 category_133 portal_5 item_type_1 month_year_09_2026

Cluster validations for adult glial annotations. Amanda A. G. Ferreira (25132828) Sergio Córdoba (645937) Raghuvanshi Rajesh (25132831) Ben Jiwon Choi (25132834) Claude Desplan (45436) Cell Biology Genetics Neuroscience Environmental Sciences not elsewhere classified Biological Sciences not elsewhere classified Developmental Biology Cancer Hematology Plant Biology innovative computational approach genetic markers expressed div >< p subcellular mrna compartmentalization optic lobe glia two large single detailed transcriptomic analysis identify mrna species glial diversity observed adult visual system glial cell bodies glial cell types understand glial development visual system cell mrna cell bodies glial types glial cells glia types adult stages cell class cell atlas neuropil glia chiasm glia work provides transcriptionally appear pupal stages helps explain gradually change drosophila </ distinct mrnas differentially localized developmental trajectories developmental single comprehensively annotated also present allowed us (A) Dot plot showing the expression levels of the genes used for the validation of each annotated cluster in adult. (B) Left, hierarchical tree showing cluster annotations after validations reveals 14 distinct cell types (in bold text) plus a number of clusters annotated preliminarily as Astrocyte- and Ensheathing-like glia. Right, unsupervised clustering of adult glial cells. (C–L) Immunostaining analysis of adult stage optic lobe. Neuropil structure is revealed by Ncad or HRP staining (yellow), and Repo (magenta) labels glia nuclei in the left panels. Marker gene expression is shown using antibodies, lacZ reporters, protein tags, or specific Gal4 lines (cyan). (C) Tret1-1-Gal4 (cyan) is only expressed in perineurial glia and not in fenestrated. (D) Vmat-Gal4 (cyan) is expressed in both perineurial and fenestrated glia. (E) rost-lacZ reporter (cyan) is expressed only in subperineurial glia. (F) GFP-tagged CG3036 (green) is expressed only in chalice glia. Vvl antibody (cyan) labels distal and proximal satellite glia. Optix (yellow) is present in chalice, marginal, epithelial, and proximal satellite glia. (G) b-Gal4 (cyan) is expressed in distal and proximal satellite glia and in epithelial glia, but not in marginal. (H) GstT4-Gal4 (cyan) is expressed only in marginal glia. (I) DAT-Gal4 (cyan) is expressed in outer chiasm glia but only seen rarely in inner chiasm glia. (J) GFP-tagged CG31663 (cyan) is expressed in proximal satellite glia and some medulla neuropil glia. This might reflect ensheathing- (mng+) vs. astrocyte-like (mng−) identity. It is also expressed in outer and inner chiasm glia and in cortex glia. (K) ome-Gal4 (cyan) is expressed in inner chiasm glia, marginal glia and some medulla neuropil glia, which could reflect ensheathing (mng+) vs. astrocytes-like (mng−) identity. (L) nkd-lacZ reporter (cyan) is expressed in medulla neuropil glia but not in lobula neuropil glia. fg = fenestrated glia, pg = perineurial glia, spg = subperineurial glia, dsg = distal satellite glia, psg = proximal satellite glia, chg = chalice glia, eg = epithelial glia, mg = marginal glia, xgo = outer chiasm glia, cxg = cortex glia, lng = lobula neuropil glia, mng = medulla neuropil glia. Gal4-UAS (>>), GFP/LacZ from construct (::). Scale bar = 20 µm. The code used to generate the graphs can be found at <a href="https://zenodo.org/records/21890456" target="_blank">https://zenodo.org/records/21890456</a>. 2026-09-28T17:57:37Z Image Figure 10.1371/journal.pbio.3004015.g003 https://figshare.com/articles/figure/_p_Cluster_validations_for_adult_glial_annotations_p_/34017682 CC BY 4.0