Journal: bioRxiv
Article Title: Membrane-anchored PrP Sc is the trigger for prion synaptotoxicity
doi: 10.1101/2025.07.11.664221
Figure Lengend Snippet: (A) Hippocampal neurons from ZH3 (Prnp -/- ) mice were transduced at DIV6 with a lentivirus encoding eGFP under control of the synapsin promoter to achieve neuron-specific expression. Cultures were then imaged on DIV21 using the EVOS Live Cell Imaging Microscope with transillumination ( Trans ) or fluorescence illumination ( GFP ). Merge shows a superposition of the two images. (B) Prnp -/- neurons were untransduced, or were transduced with lentiviruses encoding WT, G126V, or V208M PrP. Cells were fixed and stained with Alexa488-phalloidin (green) to visualize F-actin in dendritic spines and with D18 antibody (red) to detect PrP. Neurons from ZH3 (Prnp -/- ) and C57BL/6 (Prnp +/+ ) mice were used as negative and positive controls, respectively. Scale bar = 20 µm. (C) Higher magnification images of neurons showing dendritic shafts with protruding spines. Scale bar = 5 µm. (D) Dendritic spines of 12-15 neurons (each having 3-5 dendrites/neuron) from 2 independent experiments were counted in randomly selected areas, and statistical comparisons made using a one-way ANOVA multiple comparison test. Spine number is expressed per µm length of dendrite. Prnp -/- vs WT, p=0.4802; Prnp -/- vs G126V, p=0.9493; Prnp -/- vs V208M, p=0.2103. ns = not significant.
Article Snippet: Images were acquired using a Zeiss AxioObserver D1 Fluorescence Microscope and/or Zeiss LSM 700 Laser Scanning Confocal Microscope with 63X oil objectives.
Techniques: Control, Expressing, Live Cell Imaging, Microscopy, Fluorescence, Transduction, Staining, Comparison