Journal: Frontiers in Behavioral Neuroscience

Article Title: Altered cognition and anxiety in adolescent offspring whose mothers underwent different-pattern maternal sleep deprivation, and cognition link to hippocampal expressions of Bdnf and Syt-1

doi: 10.3389/fnbeh.2022.1066725

Figure Lengend Snippet: Effects of MSD on the expression of hippocampal of BDNF and Syt-1 proteins in the offspring. (A) The expression levels of BDNF ( n = 12, 6 females and 6 males in each treatment group) and Syt-1 ( n = 12, 6 females and 6 males in each treatment group) proteins in the hippocampus at 2 months. The results of protein quantification are depicted in (B,C) . ** P < 0.01 vs. Male CON group; ## P < 0.01 vs. Male CSD3h group; $$ P < 0.01 vs. Male CSD6h group; aa P < 0.01 vs. Female CON group; bb P < 0.01 vs. Female CSD3h group; cc P < 0.01 vs. Female CSD6h group.

Article Snippet: The membrane was subsequently incubated with the rabbit anti-BDNF (1:1000; Abcam, Cambridge, UK) and rabbit anti-Syt-1 (1:1000; Bioss, Beijing, China) primary antibodies, and the horseradish peroxidase (HRP) labeled goat anti-rabbit IgG (Zsbio, ZB-2301) secondary antibody, according to the manufacturer's instructions.

Techniques: Expressing

Journal: Frontiers in Behavioral Neuroscience

Article Title: Altered cognition and anxiety in adolescent offspring whose mothers underwent different-pattern maternal sleep deprivation, and cognition link to hippocampal expressions of Bdnf and Syt-1

doi: 10.3389/fnbeh.2022.1066725

Figure Lengend Snippet: The effects of MSD on the expression of hippocampal of BDNF and Syt-1 mRNA in the offspring. The mRNA expression levels of BDNF ( n = 16, 8 females and 8 males in each treatment group) and Syt1 ( n = 16, 8 females and 8 males in each treatment group) in the hippocampus at 2 months. The results of mRNA quantification are depicted in (A,B) . ** P < 0.01 vs. Male CON group; # P < 0.05 and ## P < 0.01 vs. Male CSD3h group; $ P < 0.05 and $$ P < 0.01 vs. Male CSD6h group; a P < 0.05 and aa P < 0.01 vs. Female CON group; b P < 0.05 and bb P < 0.01 vs. Female CSD3h group; c P < 0.05 and cc P < 0.01 vs. Female CSD6h group.

Article Snippet: The membrane was subsequently incubated with the rabbit anti-BDNF (1:1000; Abcam, Cambridge, UK) and rabbit anti-Syt-1 (1:1000; Bioss, Beijing, China) primary antibodies, and the horseradish peroxidase (HRP) labeled goat anti-rabbit IgG (Zsbio, ZB-2301) secondary antibody, according to the manufacturer's instructions.

Techniques: Expressing

Journal: bioRxiv

Article Title: Distinct nano-structures support a multifunctional role of actin at presynapses

doi: 10.1101/2022.05.18.492480

Figure Lengend Snippet: A-C. Anti-synaptotagmin (syt) antibody feeding experiments to assess the cycling activity of presynapses. A, cartoon of the anti-syt feeding experiments. Living neurons are incubated with the syt antibody (orange) directed against an extracellular epitope of synaptotagmin (yellow), either for 60 minutes to measure constitutive cycling, or for 3 minutes in the presence of 50 mM KCl to measure stimulated cycling. Neurons are then fixed and the syt antibody is revealed with a secondary antibody (green). B, widefield fluorescence image of cultured neurons 2 days after bead seeding at 8 div, labeled for actin (green), synaptophysin (purple), map2 (gray), and feeding with anti-synaptotagmin antibody (syt) during a 3 min incubation with KCl (stimulated cycling). C, zooms corresponding to the natural synapse (NS), S+ and S– axon-bead contacts highlighted in B. Scale bars for B, 20 µm; for C, 2 µm. D-E. FM1-43 dye loading/release experiment to assess the cycling activity of presynapses. D, cartoon of the FM1-43 experiment. Living neurons are first loaded with FM1-43 using a 3 min incubation in 50 mM KCl followed by 15 min of recovery, then images of the “loaded” time point are taken. FM1-43 is then released using a second 3-min incubation with 50 mM KCl and 15-min recovery, before images are taken of the “released” time point. E, widefield fluorescence image of cultured neurons 2 days after bead seeding at 8 div, labeled for actin using SiR (green) after loading with FM1-43 (orange). F, zooms corresponding to area highlighted in E, showing the images obtained after loading (left column) and release (right column) of FM1-43. Beads are indicated by dashed-line circles labeling the induced presynapses (S+) and axon-bead contact with no presynapse (S–). Scale bars for B, 20 µm; for C, 5 µm.

Article Snippet: Vesicular cycling assays were performed using an antibody directed to an extracellular epitope of synaptotagmin 1 (syt, clone 604.2, Synaptic Systems) that was fed to neurons 2 days after bead seeding (10 div).

Techniques: Activity Assay, Incubation, Fluorescence, Cell Culture, Labeling