Journal: bioRxiv

Article Title: Anti-Sense Oligonucleotide as a Therapeutic for Synucleinopathies: Pharmacokinetic, Safety and Efficacy Evaluation

doi: 10.1101/2025.05.01.651722

Figure Lengend Snippet: Wild-type C57BL/6 mice (n = 3) received intraperitoneal injection of biotin-labeled α-Syn ASO conjugated to ApoB 11 (2 mg/kg) to assess CNS delivery. Biotin-tagged ASO was used to visualize drug localization in brain tissue. (A) Representative immunofluorescence images showing ASO-α-Syn localization (green) in the cortex, hippocampus, substantia nigra, striatum, and thalamus. Neuronal marker NeuN (red) and nuclear stain DAPI (blue) are shown. Quantification of NeuN⁺ neurons positive for ASO-α-Syn reveals highest uptake in substantia nigra (18.5%) and hippocampus (18%), followed by striatum (14.5%), cortex (12%), and thalamus (9%). (B) Immunofluorescence staining for ASO-α-Syn (green) in CNS glial and endothelial cells, including astrocytes (GFAP, red), oligodendrocytes (Olig2, red), microglia (Iba1, red), and endothelial cells (Lectin, red). White arrows indicate ASO-positive cells. Quantification of ASO-positive cells per cell type shows highest uptake in endothelial cells (26%), followed by astrocytes (12%) and oligodendrocytes (4.5%). Microglial uptake was minimal (0.5%). Data are shown as mean ± SD

Article Snippet: Central nervous system (CNS) cells were labeled with the following antibodies: anti-NeuN (Millipore, Cat# MAB377, RRID: AB_2298772) for neurons, anti-GFAP (Millipore, Cat# MAB3402, RRID:AB_94844) for astrocytes, anti-Olig2 (Novus, Cat# NBP1-28667, RRID:AB_1914109) for oligodendrocytes, anti-Iba1 (FUJIFILM Wako Pure Chemical Corporation Cat# 019-19741, RRID:AB_839504) for microglia, and Tomato-lectin (Bioworld, Cat# 21761030-1, RRID:AB_2336416) for endothelial cells.

Techniques: Injection, Labeling, Immunofluorescence, Marker, Staining

Journal: Glia

Article Title: The X-Linked Intellectual Disability Gene, ZDHHC9, Is Important for Oligodendrocyte Subtype Determination and Myelination.

doi: 10.1002/glia.70016

Figure Lengend Snippet: FIGURE 2 | There is a change in mature oligodendrocyte subtype densities, and an increase in the density of OPCs in the corpus callosum of Zdhhc9-KO mice. (a) Representative images of fluorescent in situ hybridization for MOL2/3 and MOL5/6 (Hapln2—MOL2/3; C030029H02Rik— MOL5/6; Ptgds—MOL5/6) in the P60 corpus callosum (scale bar, 50 μm) (b) Representative image of corpus callosum cells colored by oligodendro- cyte subtype classification. MOL2/3 cells are colored green; MOL5/6 cells are colored blue; unclassified cells are colored gray. c Graph of oligoden- drocyte subtype proportions (n = 5 mice per genotype). (d) Representative images of fluorescent in situ hybridization for immature oligodendrocyte cell-lineage-subtype marker genes (Pdgfra-OPCs, Itpr2-NFOLs, Ctps-MFOLs) in the corpus callosum (scale bar, 50 μm). (e) Representative image of corpus callosum cells colored by oligodendrocyte subtype classification. OPC cells are colored yellow; NFOL cells are colored cyan; MFOL cells are colored magenta; unclassified cells are colored gray. (f) Graph of OPC, NFOL and MFOL proportions (n = 4 Zdhhc9-KO mice, n = 5 control mice). (g) Representative images of OLIG2 (pan oligodendrocyte marker) and CC1 (mature oligodendrocyte marker) immunolabeling in the P60 corpus callo- sum. h Graph of OLIG2+ and OLIG2+/CC1+ proportions (n = 5 mice per genotype). Data in (c, f, h) show mean ± SEM. (c, f, h) multiple t-tests using Holm-Sidak method, alpha = 0.05.

Article Snippet: See the T erm s and C onditions (https://onlinelibrary.w iley.com /term s-and-conditions) on W iley O nline L ibrary for rules of use; O A articles are governed by the applicable C reative C om m ons L icense 4 of 15 Glia, 2025 (1:500, NBP1- 28667, Novus) and Ki67 (1:500, ab15580, Abcam); or Olig2 (1:500, NBP1- 28667, Novus) and CC3 (1:500, 9661S, Cell Signaling Techno) were incubated overnight at 4°C.

Techniques: In Situ Hybridization, Marker, Control, Immunolabeling