Journal: bioRxiv

Article Title: Neural circuit basis of adolescent THC-induced potentiation of opioid responses in adult mice

doi: 10.1101/2024.04.23.590773

Figure Lengend Snippet: A , Representative sagittal section of a cFos-stained brain of an animal treated with THC during adolescence and following a reinstatement dose of morphine. Scale, 2 mm. B , More cFos-expressing cells were detected in adolescent THC-treated mice than vehicle-treated mice following a reinstatement dose of 5 mg/kg morphine (548,481 vs. 325,789 cells, p = 0.0051). C , The top ten brain regions showing the highest-fold elevation in cFos labeling in adolescent THC-treated relative to vehicle-treated mice and the four regions showing an elevation in vehicle-treated mice. Groups are separated by the dashed red line. Shown are brain regions with an average > 150 cFos+ cells in each brain regions. D , Representative sagittal images of the GPe in adolescent vehicle-treated vs. THC-treated mice. Scale, 500 μm. E , Anatomical connectivity matrix of the mouse brain, broken into four communities based on the Louvain method for community detection F , Log fold-change in cFos labeling in each network community relative to one another (One-way ANOVA p = 0.0002; purple (1) vs. blue (2) 0.68 vs. 0.75, p = 0.76; purple (1) vs. red (3) 0.68 vs. 0.61, p = 0.82; purple (1) vs. yellow (4) 0.68 vs. 0.38, p = 0.0024; blue (2) vs. red (3) 0.75 vs. 0.61, p = 0.31; blue (2) vs. yellow (4) 0.68 vs. 0.38, p = 0.0002; red (3) vs. yellow (4) 0.61 vs. 0.38, p = 0.05. G , UMAP embedding of regions based on their correlated activity in adolescent vehicle-treated mice. H , UMAP embedding of regions based on their correlated brain activity in adolescent THC-treated mice. I , Hierarchical clustering of regions based on the cFos correlation matrix in adolescent vehicle-treated mice. J , Hierarchical clustering of regions based on the cFos correlation matrix in adolescent THC-treated mice. K , The number of hierarchical clusters in adolescent vehicle-treated and THC-treated mice across a range of heights for cluster cut-offs in hierarchical trees.

Article Snippet: The monoclonal rat anti-cFos antibody (Synaptic Systems lot #226017) in 5% NDS/PBS-Tween 0.1% was used at a 1:5000 dilution, with an incubation time of five days.

Techniques: Staining, Expressing, Labeling, Activity Assay

Journal: Stem cells (Dayton, Ohio)

Article Title: Retinoic Acid-signaling regulates the proliferative and neurogenic capacity of Müller glia-derived progenitor cells in the avian retina

doi: 10.1002/stem.2742

Figure Lengend Snippet: Antibodies, sources and working dilutions. Patterns of labeling and stimulus-dependent changes in levels of immunolabeling using these antibodies are consistent with previous reports (Fischer and Omar, 2005; Fischer et al., 2009a , 2009b ; Fischer et al., 2014 ; Todd and Fischer, 2015 ).

Article Snippet: Secondary antibodies included donkey-anti-goat-Alexa488/568, goat-anti-rabbit-Alexa488/568/647, goat-anti-mouse-Alexa488/568/647, goat anti-rat-Alexa488 (Life Technologies) diluted to 1:1000 in PBS plus 0.2% Triton X-100. table ft1 table-wrap mode="anchored" t5 Table 1 caption a7 Antigen Working dilution Host Clone or catalog number Source Sox2 1:1000 goat Y-17 Santa Cruz Immunochemicals Sox9 1:2000 mouse AB5535 Chemicon Pax6 1:1000 rabbit PRB-278P Covance Klf4 1:50 rabbit {"type":"entrez-protein","attrs":{"text":"ARP38430","term_id":"1190169341","term_text":"ARP38430"}} ARP38430 Aviva Systems Biology cFos 1:400 rabbit K-25 Santa Cruz Immunochemicals Glutamine Synthetase 1:2000 mouse ab125724 Abcam CD45 1:200 Mouse HIS-C7 Cedi Diagnostic Nkx2.2 1:80 Mouse 74.5A5 DSHB CRABP 1:1000 Mouse C1 Dr. J Saari, University of Washington Lim 1/2 1:50 Mouse 4F2 DSHB Lim 3 1:100 Mouse 67.4E12 DSHB Otx2 1:1000 Goat AF1979 R&D Systems HuD/HuC 1:600 Mouse A21271 Invitrogen Open in a separate window Antibodies, sources and working dilutions.

Techniques: Labeling, Immunolabeling, Diagnostic Assay

Journal: Molecular Psychiatry

Article Title: Age, sex, and apolipoprotein E isoform alter contextual fear learning, neuronal activation, and baseline DNA damage in the hippocampus

doi: 10.1038/s41380-023-01966-8

Figure Lengend Snippet: A Amount of DNA pulled down by γH2Ax ChIP in naïve females. There was an effect of age ( p < 0.001, F (1,18) = 14.501) and genotype ( p < 0.001, F (2,18) = 29.522). E4 mice had more γH2Ax-associated DNA than E2 ( p < 0.0001) or E3 ( p < 0.0001). B Amount of DNA pulled down by γH2Ax ChIP in naive males. There was an effect of age ( p = 0.002, F (1,18) = 13.321) and an age-by-genotype interaction ( p = 0.021, F (2,18) = 4.847). C Number of cells with 53BP1 foci in the CA3 of naïve females. Middle-aged animals had more 53BP1 than young animals ( p = 0.029, F (1, 13) = 6.036). D Number of cells with 53BP1 foci in the CA3 of naïve males. There were no effects. E Relative Fos expression in the hippocampus of naïve females. Middle aged animals had more than young animals ( p = 0.008, F (1,16) = 9.082). F Relative Fos expression in the hippocampus of naïve males. There were no effects. G Number of cells with 53BP1 foci in the CA3 of naïve females. There was an effect of age ( p = 0.0003, F (1,13) = 24.16) and genotype ( p = 0.045, F (2,13) = 3.975). H Number of cells with 53BP1 foci in the CA3 of naive males. There was an effect of genotype ( p = 0.025, F (2, 13) = 4.941) and an age-by-genotype interaction ( p = 0.023, F (2,13) = 5.096). Middle-aged E2 had more than middle-aged E3 ( p = 0.004) and E4 ( p = 0.007) mice. I cFos in the CA1 of young females. There was an effect of time post-training ( p < 0.0001, F (2, 22) = 67.61); mice euthanized at 1.5 h had mor cFos than in behaviorally naïve or 24 h for all genotypes. J cFos in the CA1 of young males. There was an effect of time post-training ( p = 0.015, F (2,20) = 5.233); post hoc testing did not reach significance. K cFos in the CA1 of middle-aged females. There was an effect of time post-training ( p = 0.011, F (2,20) = 5.656) and a trend towards an effect of genotype ( p = 0.063). Only E2 mice euthanized at 1.5 h had higher cFos than behaviorally naïve or 24 h. L cFos in the CA1 of middle-aged males. There was an effect of time post-training ( p < 0.0001, F (2,21) = 29.38) and genotype ( p < 0.0001, F (2,21) = 37.70) and a time-by-genotype interaction ( p < 0.001, F (4,21) = 8.85). Only E2 mice euthanized at 1.5 h had higher cFos than behaviorally naive or 24 h. M Representative images of the CA1. White arrows indicate cFos+ cells, yellow arrows indicate 53BP1 + cells. Right : zoomed images of the white boxes. Data are presented as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, ^ p < 0.0001.

Article Snippet: Immunofluorescence was done on free-floating 40 μm sections [ , ] using primary antibodies against 53BP1 (Bethyl Labs, #A300-272A) and cFos (PhosphoSolutions, #309-cFOS), secondary antibodies (AlexaFluor647, Invitrogen, #A-21245; and AlexaFluor488, Invitrogen, #A-11001) and a DAPI (Sigma D9542) counterstain.

Techniques: Expressing