Journal: bioRxiv
Article Title: Repetitive Neuronal Activation Regulates Cellular Maturation State via Nuclear Reprogramming
doi: 10.1101/2025.05.02.651848
Figure Lengend Snippet: a , Simultaneous in vivo calcium imaging and optogenetic stimulation under free-moving conditions. AAV-syn-GCaMP6f and AAV-syn-ChrimsonR were co-injected into the DG, followed by GRIN-lens implantation. An integrated miniature microscope (nVoke; Inscopix) enabled large-scale cellular-resolution Ca 2+ imaging and optogenetic stimulation. b, Left: Expression of GCaMP6f and ChrimsonR in the DG. Right: Max projection of relative fluorescence change (Δ F / F ) of Ca 2+ transients from all imaging frames during the first 30 min, showing representative active neurons. c, Top: Experimental design of Ca 2+ imaging combined with optogenetic stimulation during the open field test. Each 50-min session consisted of 30-min Ca 2+ imaging (used for the following analysis), 5-min optogenetic stimulation, and 15-min post-stimulation recording. This was repeated for 10 days. No light was introduced to mice in the No Stim group. Bottom: Δ F / F traces from 15 representative neurons (scale bar, 10% Δ F / F ). Data from 25 to 38 min of a 50-min session are shown. d, Distance traveled during the first 30 min e, Average Ca 2+ transient rate during the first 30 min. f, Distribution of spatial information for the No Stim (grey curve) and Stim×10 groups (red curve). The vertical axis represents the frequency of distribution, with the dotted line indicating the criterion for place cells (top 95% percentile of the shuffled distribution; see Methods section). Bar graph indicates proportion of place cells. g, Representative results of position decoding using Ca 2+ imaging data. The first 30 min was split into two 15-min halves for training and test data for decoding. Black/red lines: observed position; grey/pink lines: decoded position. h, Decoding accuracy (mae; mean absolute error, cm). Dotted lines: shuffled control. Two-way repeated measures ANOVA: Stim type, F (1, 8) = 6.49, P = 0.034; Day, F (2, 16) = 2.89, P = 0.085; Stim type × Day, F (2, 16) = 1.11, P = 0.353. Bonferroni correction for multiple comparisons was performed, * P < 0.05. i, Same as f , but for speed information and speed cells. The dotted line indicates the criterion for speed cells (top 99% percentile of the shuffled distribution). j, Same as g , but for speed decoding. k, Same as h , but for speed decoding accuracy. Two-way repeated measures ANOVA: Stim type, F (1, 8) =4.02, P = 0.080; Day, F (2, 16) = 4.87, P = 0.022; Day×Stim type, F (2, 16) = 3.98, P = 0.039. Bonferroni correction for multiple comparisons was performed, * P < 0.05.
Article Snippet: To extract the activity patterns of individual DG neurons from the obtained fluorescent images, we used Inscopix Data Processing Software (IDPS 1.8.0).
Techniques: In Vivo, Imaging, Injection, Microscopy, Expressing, Fluorescence, Control
