Journal: bioRxiv

Article Title: A Single-Cell Signaling Atlas of Spinal Cord BDNF Responses Reveals Determinants Beyond Receptor Expression

doi: 10.64898/2026.03.24.713745

Figure Lengend Snippet: ( A ) Embryonic day 14 (E14) rat spinal cord are cultured to day in vitro (DIV) 3.5 before a 12hr supplement starvation and subsequent treatment. Samples are collected at indicated time points by rapid dissociation (<90s) and fixation by 2% paraformaldehyde (PFA), then stored at -80°C. Samples are barcoded, pooled, and simultaneously stained with isotopically pure metal-conjugated antibodies before and after methanol (MeOH) permeabilization and fixation. Stained samples are then analyzed by single-cell (suspension) mass cytometry. Created using BioRender. ( B ) Leiden clustering by cell identity markers is visualized by dimensionality reduction on a 3-Dimensional Uniform Manifold Association Plot (UMAP) colored and labeled by identified cell types. ( C ) The resulting UMAP is colored by the level of individual cell identity markers to highlight the observed neuronal maturation gradient that shifts from Sox2+, BLBP+ progenitors to more mature neuronal identity with increasing levels of ßIII-Tubulin, MAP2, and NeuN. ( D ) Non-neuronal cell identity marker levels are plotted to define glial cell populations. Individual cell identity marker expression is normalized from 0 to 1, and gradient is shifted to emphasize locations of marker enrichment, with cells above the intensity threshold colored cyan.

Article Snippet: Primary dissociated neural cultures were prepared from E14 Sprague Dawley rat spinal cords.

Techniques: Cell Culture, In Vitro, Staining, Single Cell, Suspension, Mass Cytometry, Labeling, Marker, Expressing