Journal: bioRxiv
Article Title: Neurospheres from primary rodent brain cells to probe the 3D organization and function of synapses
doi: 10.64898/2026.03.19.712855
Figure Lengend Snippet: (A) Neurospheres were formed by mixing non-electroporated and electroporated cells before being plated in U-bottom ULA wells. (B) Maximum intensity projection image of a confocal stack of a DIV14 neurosphere in which neurons were separately electroporated with RFP (red) and Xph20-GFP (green). (C) Zoomed images on primary dendrites from neurons expressing intrabodies to PSD-95 (Xph20-GFP), or gephyrin (GPHN.FingR-GFP), revealing excitatory or inhibitory post-synapses, respectively. (D) Numbers of PSD-95 and gephyrin-positive puncta per electroporated neuron. Data represent the mean ± SEM of 14 and 10 neurospheres, respectively, and were compared by non-parametric Mann-Whitney test. Dots show individual neurospheres. (E) Maximum intensity projection of a confocal stack of a DIV14 neurosphere in which neurons were co-electroporated with GPHN.FingR-GFP (green) and Xph20-mRuby2 (magenta), allowing the detection of both excitatory and inhibitory post-synapses in the same cells. (F) Zoom on a dendritic segment corresponding to the rectangular area highlighted in (E). (G) Confocal image of a primary dendrite from a neuron expressing GFP-actin, further immunolabeled for GFP, showing numerous dendritic spines bulging out of the shaft.
Article Snippet: The intrabody to gephyrin, GPHN.FingR-GFP (Addgene # 46296 pCAG_GPHN.FingR-eGFP-CCR5TC), was a gift from D. Arnold (University of Southern California, Los Angeles, CA, USA) ( ).
Techniques: Expressing, MANN-WHITNEY, Immunolabeling