Journal: bioRxiv
Article Title: Sleep neural code perpetuates the evolving negativity bias under stress
doi: 10.64898/2026.03.03.709197
Figure Lengend Snippet: a. Schematic of longitudinal Ca 2+ imaging recording schedules from the SoD experience session to the CFC memory recall session. b. Expression of the Janelia variant Ca 2+ indicator AAV9-hSyn-JGCaMP7b and the location of the implanted GRIN lens in the CA1 region of the dorsal hippocampus. Scale bar, 250 µm. c. Example image of cells extracted from Ca 2+ imaging and representative traces of z scored neural activity. d. Schematic of the analysis flowchart for detecting recall-specific ensembles using PCA/ICA dimensionality reduction methods. After all PCA/ICA ensembles from the CFC recall session were detected, these ensembles were further categorized into recall-specific (pink) or recall-nonspecific ensembles (blue) by comparing the ensemble activity in the CFC recall session with that in the previous CFC pre-shock session by Wilcoxon rank sum methods. For each example trace of the ensemble activity, the corresponding raster plot of the neural activity is shown with the same colour. e. Multisession comparison of the averaged activity index of the recall-specific ensembles, which is normalized by the average activity rate of the recall-nonspecific ensembles per mouse for each corresponding session. (Neutral n = 14; SoD n = 28 ensembles; two-way RM ANOVA with correction; interaction P <0.001; F (6, 240) = 5.382; Šídák’s multiple comparisons test). f. Pearson correlation analysis of the activity index of recall-specific ensemble during the SI-ICR session and prospective total freezing level (%) in the CFC recall session per SoD mouse. ( r = 0.9378, P = 0.0184, n = 5 mice) g. Activity rate of each CFC recall-specific ensemble in the neutral group in the home cage before the neutral experience (Pre-Neutral-home cage) and during the neutral session (novel home cage exploration) on day 6 ( n = 14 ensembles; Wilcoxon signed rank test; P = 0.3306); additionally, the activity rate of the SoD group in the home cage before the SoD session (Pre-SoD-home cage) and during the SoD session on day 6 of the SoD experience ( n = 28 ensembles, Wilcoxon signed rank test, P = 0.0014). h. Activity rate of each recall-specific ensemble during the novel B6 mouse encounter (SI-B6) and novel ICR mouse encounter (SI-ICR) in the social interaction test for the neutral group ( n = 14 ensembles, Wilcoxon signed rank test, P = 0.4352) and SoD group ( n = 28 ensembles, Wilcoxon signed rank test, P = 0.0066). i. Activity rate of each recall-specific ensemble during the 2-minute exploration period before receiving the first electric shock (CFC pre-shock) and during the remaining session after the delivery of the first shock (CFC post-shock) on the CFC conditioning day in the neutral group ( n = 14 ensembles; Wilcoxon signed rank test; P = 0.0076) and the SoD group ( n = 28 ensembles, Wilcoxon signed rank test, P = 0.0053). j. Schematic for assessing the correlation of the CFC recall-specific ensemble activity during opposite corner stays during the SI-ICR session and the ensemble activity during the freeze state in the CFC recall session. Example traces of averaged recall-specific ensemble activity (z scored) around the onset of the opposite corner entries in the SI-ICR (lower left) and during freezing behaviours in CFC recall (lower right) sessions. k. Spearman correlation ( r value shown) for the activity of the CFC recall-specific ensembles in the neutral group during opposite corner stays and activity during freezing in the CFC recall session for the neutral group ( r = - 0.3600, P = 0.2047) and l. SoD group ( r = 0.4426, P = 0.0208, n = 27 ensembles). m. Representative heatmaps showing the intercell type pairwise correlation analysis between SI avoidance-related (SI opposite corner-related neurons) and CFC aversion-related (shock- and freeze-related neurons) cells in the non-freezing state (left) and the freezing state in the CFC recall session (right). Red indicates a significant correlation between two neurons. n. Percentage of significant pairwise correlations of SI avoidance-related and CFC aversion-related cells during the freezing state in the CFC recall session (Neutral n = 5; SoD n = 5 mice; unpaired t test, t = 2.653; P = 0.0291). o. Percentage of significant intracell type neuron‒neuron pairwise correlations of SI avoidance-related neurons during the freezing state in the CFC recall session (Neutral, n = 5; SoD, n = 5 mice; Unpaired t test, t = 0.2808; P = 0.786). Data are presented as the mean ± s.e.m. Statistical significance is expressed as * P < 0.05; ** P < 0.01; and *** P < 0.001.
Article Snippet: For longitudinal in vivo Ca2+ imaging experiments, we used a recombinant adeno-associated virus (AAV) vector, AAV9-hSyn-jGCaMP7b (Janelia variant, Addgene: #104489, viral titer: 9.77 × 1013 vg/mL).
Techniques: Imaging, Expressing, Variant Assay, Activity Assay, Comparison
Addgene inc is a verified supplier 
